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Sample records for fertilization in vitro

  1. IVF-In Vitro Fertilization.

    ERIC Educational Resources Information Center

    Kieffer, George H.

    1980-01-01

    Issues surrounding the controversial topic of in vitro fertilization and artificial manipulation of reproduction are discussed. The author examines the moral and ethical implications and presents results of a survey of various religious groups. (SA)

  2. In vitro fertilization (IVF)

    MedlinePlus

    ... Assisted reproductive technology; ART; Test-tube baby procedure; Infertility - in vitro ... It is used to treat many causes of infertility , including: Advanced age of the woman (advanced maternal ...

  3. Human rights to in vitro fertilization.

    PubMed

    Zegers-Hochschild, Fernando; Dickens, Bernard M; Dughman-Manzur, Sandra

    2013-10-01

    The Inter-American Court of Human Rights (the Court) has ruled that the Supreme Court of Costa Rica's judgment in 2000 prohibiting in vitro fertilization (IVF) violated the human right to private and family life, the human right to found and raise a family, and the human right to non-discrimination on grounds of disability, financial means, or gender. The Court's conclusions of violations contrary to the American Convention on Human Rights followed from its ruling that, under the Convention, in vitro embryos are not "persons" and do not possess a right to life. Accordingly, the prohibition of IVF to protect embryos constituted a disproportionate and unjustifiable denial of infertile individuals' human rights. The Court distinguished fertilization from conception, since conception-unlike fertilization-depends on an embryo's implantation in a woman's body. Under human rights law, legal protection of an embryo "from conception" is inapplicable between its creation by fertilization and completion of its implantation in utero.

  4. Antisperm antibodies and in vitro fertilization failure.

    PubMed

    Pexieder, T; Boillat, E; Janecek, P

    1985-12-01

    A new enzyme-linked immunosorbent assay (ELISA)-based test (Zer, Jerusalem) has allowed us to show the presence of antisperm antibodies (1:32 to 1:64) in the blood of 14 (33%) of 32 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) under gonadotropin stimulation. Observation of the morphology of fertilization in eight patients with and seven patients without antisperm antibodies has shown a significant association (P less than or equal to 0.025, Fisher exact probability test) among the cumulus/corona coagulation, the absence of fertilization, and the presence of these antibodies. Cumulus/oocyte complex washing and/or enzymatic cumulus removal are considered as elective interventions in the case of antisperm immunity. Each patient entering an IVF-ET program should have the antisperm antibody assay performed as a preliminary screening.

  5. [Circulating nucleic acids and in vitro fertilization].

    PubMed

    Scalici, E; Traver, S; Mullet, T; Ferrières, A; Monforte, M; Vintejoux, E; Hamamah, S

    2014-10-01

    During the last years, the use of circulating nucleic acids (microRNAs and cell-free DNA) as diagnostic and/or prognostic tools in cancerology was widely documented. Likewise, in obstetrics and gynecology, the development of non-invasive prenatal testing based on the assessment of these biomarkers confirmed their growing interest in this speciality. In human reproduction, several studies were interested in the microRNAs, small non-coding RNA sequences, present in the ovarian follicle and their implication in folliculogenesis. Some of these microRNAs, as well as the vesicles which transport them, are easily detectable in the bloodstream and could be used as reliable biomarkers of interest in infertility care. Cell-free DNA level varies according to physiopathology and reflect the proportion of apoptotic and/or necrotic events occurring in the body. As a result, its quantification could give an additional help to the practitioners for ovarian functional status evaluation. Furthermore, these circulating nucleic acids could also constitute new predictive biomarkers of oocyte and/or embryo quality and represent a promising perspective for the prevention of in vitro fertilization implantation failures. In conclusion, these circulating nucleic acids open the way to the development of new diagnostic and/or prognostic innovative tests in order to improve in vitro fertilization outcomes.

  6. Mild stimulation in in vitro fertilization.

    PubMed

    Macklon, N S; Fauser, B C J M

    2003-11-01

    Current approaches to ovarian stimulation for in vitro fertilization (IVF) are aimed at optimizing the number of oocytes retrieved in a treatment cycle. This approach is not without risks. Moreover, as the true costs of multiple pregnancy become clearer, the need to produce multiple embryos for transfer is increasingly questioned. Increasing knowledge of the physiological mechanisms involved in follicular development and dominance has led to new strategies in ovarian stimulation for IVF. The clinical availability of GnRH antagonists allows the normal cycle to be harnessed and manipulated by mild interventions to produce sufficient oocytes for successful IVF treatment. Recent evidence suggests that oocyte quality after mild stimulation may be superior to that after conventional stimulation regimens.

  7. Cell adhesion molecules and in vitro fertilization.

    PubMed

    Simopoulou, Maria; Nikolopoulou, Elena; Dimakakos, Andreas; Charalabopoulos, Konstantinos; Koutsilieris, Michael

    2014-01-01

    This review addresses issues regarding the need in the in vitro fertilization (IVF) field for further predictive markers enhancing the standing embryo selection criteria. It aims to serve as a source of defining information for an audience interested in factors related to the wide range of multiple roles played by cell adhesion molecules (CAMs) in several aspects of IVF ultimately associated with the success of an IVF cycle. We begin by stressing the importance of enriching the standing embryo selection criteria available aiming for the golden standard: "extract as much information as possible focusing on non-invasive techniques" so as to guide us towards selecting the embryo with the highest implantation potential. We briefly describe the latest trends on how to best select the right embryo, moving closer towards elective single embryo transfer. These trends are: frozen embryo transfer for all, preimplantation genetic screening, non-invasive selection criteria, and time-lapse imaging. The main part of this review is dedicated to categorizing and presenting published research studies focused on the involvement of CAMs in IVF and its final outcome. Specifically, we discuss the association of CAMs with conditions and complications that arise from performing assisted reproductive techniques, such as ovarian hyperstimulation syndrome, the state of the endometrium, and tubal pregnancies, as well as the levels of CAMs in biological materials available in the IVF laboratory such as follicular fluid, trophectoderm, ovarian granulosa cells, oocytes, and embryos. To conclude, since CAMs have been successfully employed as a diagnostic tool in several pathologies in routine clinical work, we suggest that their multi-faceted nature could serve as a prognostic marker in assisted reproduction, aiming to enrich the list of non-invasive selection and predictive criteria in the IVF setting. We propose that in light of the well-documented involvement of CAMs in the developmental

  8. Cell adhesion molecules and in vitro fertilization.

    PubMed

    Simopoulou, Maria; Nikolopoulou, Elena; Dimakakos, Andreas; Charalabopoulos, Konstantinos; Koutsilieris, Michael

    2014-01-01

    This review addresses issues regarding the need in the in vitro fertilization (IVF) field for further predictive markers enhancing the standing embryo selection criteria. It aims to serve as a source of defining information for an audience interested in factors related to the wide range of multiple roles played by cell adhesion molecules (CAMs) in several aspects of IVF ultimately associated with the success of an IVF cycle. We begin by stressing the importance of enriching the standing embryo selection criteria available aiming for the golden standard: "extract as much information as possible focusing on non-invasive techniques" so as to guide us towards selecting the embryo with the highest implantation potential. We briefly describe the latest trends on how to best select the right embryo, moving closer towards elective single embryo transfer. These trends are: frozen embryo transfer for all, preimplantation genetic screening, non-invasive selection criteria, and time-lapse imaging. The main part of this review is dedicated to categorizing and presenting published research studies focused on the involvement of CAMs in IVF and its final outcome. Specifically, we discuss the association of CAMs with conditions and complications that arise from performing assisted reproductive techniques, such as ovarian hyperstimulation syndrome, the state of the endometrium, and tubal pregnancies, as well as the levels of CAMs in biological materials available in the IVF laboratory such as follicular fluid, trophectoderm, ovarian granulosa cells, oocytes, and embryos. To conclude, since CAMs have been successfully employed as a diagnostic tool in several pathologies in routine clinical work, we suggest that their multi-faceted nature could serve as a prognostic marker in assisted reproduction, aiming to enrich the list of non-invasive selection and predictive criteria in the IVF setting. We propose that in light of the well-documented involvement of CAMs in the developmental

  9. Cost analysis of in vitro fertilization.

    PubMed

    Stern, Z; Laufer, N; Levy, R; Ben-Shushan, D; Mor-Yosef, S

    1995-08-01

    In vitro fertilization (IVF) has become a routine tool in the arsenal of infertility treatments. Assisted reproductive techniques are expensive, as reflected by the current "take home baby" rate of about 15% per cycle, implying the need for repeated attempts until success is achieved. Israel, today is facing a major change in its health care system, including the necessity to define a national package of health care benefits. The issue of infertility and whether its treatment should be part of the "health basket" is in dispute. Therefore an exact cost analysis of IVF is important. Since the cost of an IVF cycle varies dramatically between countries, we sought an exact breakdown of the different components of the costs involved in an IVF cycle and in achieving an IVF child in Israel. The key question is not how much we spend on IVF cycles but what is the cost of a successful outcome, i.e., a healthy child. This study intends to answer this question, and to give the policy makers, at various levels of the health care system, a crucial tool for their decision-making process. The cost analysis includes direct and indirect costs. The direct costs are divided into fixed costs (labor, equipment, maintenance, depreciation, and overhead) and variable costs (laboratory tests, chemicals, disposable supplies, medications, and loss of working days by the couples). The indirect costs are the costs of premature IVF babies, hospitalization of the IVF pregnant women in a high risk unit, and the cost of complications of the procedure. According to our economic analysis, an IVF cycle in Israel costs $2,560, of which fixed costs are about 50%. The cost of a "take home baby" is $19,267, including direct and indirect costs.

  10. Impaired fertilizing ability of superoxide dismutase 1-deficient mouse sperm during in vitro fertilization.

    PubMed

    Tsunoda, Satoshi; Kawano, Natsuko; Miyado, Kenji; Kimura, Naoko; Fujii, Junichi

    2012-11-01

    The oxidative modification of gametes by a reactive oxygen species is a major deleterious factor that decreases the successful rate of in vitro fertilization. Superoxide dismutase 1 (SOD1) plays a pivotal role in antioxidation by scavenging the superoxide anion, and its deficiency causes infertility in female mice, but the significance of the enzyme in male mice remains unclear. In the present study, we characterized Sod1(-/-) (Sod1-KO) male reproductive organs and compiled the first report of the impaired fertilizing ability of Sod1-KO sperm in in vitro fertilization. Insemination of wild-type oocytes with Sod1-KO sperm exhibited lower rates of fertility compared with insemination by wild-type sperm. The low fertilizing ability found for Sod1-KO sperm was partially rescued by reductant 2-mercaptoethanol, which suggested the oxidative modification of sperm components. The numbers of motile and progressive sperm decreased during the in vitro fertilization process, and a decline in ATP content and elevation in lipid peroxidation occurred in the Sod1-KO sperm in an incubation time-dependent manner. Tyrosine phosphorylation, which is a hallmark for sperm capacitation, was also impaired in the Sod1-KO sperm. These results collectively suggest that machinery involved in sperm capacitation and motility are vulnerable to oxidative damage during the in vitro fertilization process, which could increase the rate of inefficient fertilization.

  11. Hysteroscopy in a program of in vitro fertilization.

    PubMed

    Goldenberg, M; Bider, D; Ben-Rafael, Z; Dor, J; Levran, D; Oelsner, G; Mashiach, S

    1991-12-01

    Two hundred twenty-four women underwent hysteroscopic evaluation without anesthesia after at least two failed attempts of in vitro fertilization and embryo transfer. One hundred fifty-three (68%) women were diagnosed as having mechanical infertility, and abnormal hysteroscopic findings were observed in 32 (21%). Forty-one women were diagnosed as having unexplained infertility (18%) and six (15%) had abnormal findings with hysteroscopy. Of the 30 couples who entered the in vitro fertilization regimen program because of male infertility, 4 (13%) had abnormal findings. The overall rate of abnormal findings was 19%; cervical canal and intrauterine abnormalities were found in 10 and 32 patients, respectively. Ten patients were treated during hysteroscopic evaluation procedure, and four patients subsequently underwent operative hysteroscopy under general anesthesia. We suggest that diagnostic hysteroscopy should be a routine procedure before in vitro fertilization and embryo transfer therapy.

  12. [In vitro assessment of semen for the prediction of fertility].

    PubMed

    Waberski, D; Petrounkina, A; Weitze, K F; Töpfer-Petersen, E

    1999-02-01

    The prediction of fertility is a primary goal in the field of reproductive medicine. The aim of the present paper is to describe the value of conventional and modern sperm analysis systems considering the process of fertilization. The classical assessment of motility and morphology enables the rough estimation of semen quality in order to select ejaculates for the use in artificial insemination. Recent methods for sperm diagnosis, such as fluorescent marking for the detection of sperm plasma membrane integrity, the hypoosmotic swelling test, and computer assisted semen analysis allow for the evaluation of a large number of spermatozoa and the assessment of sperm dynamics under in vitro-fertilization conditions. The oocyte penetration test investigates the ability of spermatozoa for capacitation, hyperactivation and acrosome reaction in vitro. The amount of specific seminal plasma proteins is related to fertility and thereby provides an additional semen evaluation method. For the use of a given semen test the specific in vitro condition has to be considered. In addition, the evaluated criteria relevant for the process of fertilization need to be defined. The combination of selected semen tests gives a higher accuracy for the prediction of fertilizing capacity compared with a single test. PMID:10077807

  13. In vitro maturation and in vitro fertilization of mouse oocytes and preimplantation embryo culture.

    PubMed

    Kidder, Benjamin L

    2014-01-01

    Epigenetic regulation of gene expression in the germline is important for reproductive success of mammals. Misregulation of genes whose expression is correlated with reproductive success may result in subfertility or infertility. To study epigenetic events that occur during oocyte maturation and preimplantation embryo development, it is important to generate large numbers of ovarian follicles and embryos. Oocyte maturation can be modeled using in vitro maturation (IVM), which is a system of maturing ovarian follicles in a culture dish. In addition, fertilization and early embryogenesis can be modeled using in vitro fertilization (IVF), which involves the fertilization of mature oocytes with capacitated sperm in a culture dish. Here, we describe protocols for in vitro maturation (IVM) and in vitro fertilization (IVF) of mouse oocytes and preimplantation embryo culture. These protocols are suitable for the study of oocyte and embryo biology and the production of embryonic mice. PMID:24743999

  14. Toxicity testing for human in vitro fertilization programs.

    PubMed

    Ackerman, S B; Stokes, G L; Swanson, R J; Taylor, S P; Fenwick, L

    1985-09-01

    Using a mouse embryo culture system, several procedures and materials associated with human in vitro fertilization protocols were tested for potential toxicity. Also, quality-control assays were performed for media prepared by nine different human in vitro fertilization programs. Detrimental effects upon embryo development were observed when culture media were exposed to the following substances: surgical instruments sterilized with Cidex or Cidex-7 or sterilized with ethylene oxide after packaging in Nest Protector Packs, various brands of surgical gloves, and various synthetic materials being evaluated as possible needle or catheter "liners." Results from comparative testing of media and serum supplements prepared by different in vitro programs indicated a wide range in culture medium quality, as assayed by the ability of the reagents to support mouse embryo development. The importance of an animal model system available to all human in vitro programs for routine quality-control analysis and testing of novel uses of materials and innovative methods is discussed.

  15. Hyaluronidase removal of the cumulus oophorus increases in vitro fertilization.

    PubMed

    Lavy, G; Boyers, S P; DeCherney, A H

    1988-10-01

    The effect of hyaluronidase removal of the cumulus oophorus on the in vitro fertilization rate of oocytes obtained from patients with poor oocyte fertilizability has been evaluated. Eighty-eight oocytes were obtained from 13 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) for indications of male-factor, immunological, and idiopathic infertility. In addition, patients in whom fertilization did not occur on previous IVF cycles were evaluated in the study. The oocytes of each individual patient were randomly assigned into a treatment (removal of the cumulus; N = 40 oocytes) or nontreatment group (control; N = 48 oocytes). Hyaluronidase was used to remove the cumulus immediately following oocyte retrieval, and insemination was performed 6-8 hr later. The overall oocyte fertilization rate (both treated and untreated) was 42%. The treatment group demonstrated a higher rate of fertilization compared to the nontreatment group (55% vs 31%; P less than 0.05). Examination of various patient groups revealed a statistically significant difference in fertilization rates between the treated and the untreated oocytes only in the "no previous fertilization" group (60% vs 28%; P less than 0.05). A higher rate of fertilization of the treated oocytes was also seen in the immunologic infertility group, however, statistical significance was not achieved (50% vs 25%; P = 0.07). Only one clinical pregnancy was achieved in this group of 13 patients. We conclude that in this group of patients, removal of the cumulus prior to insemination may, in some cases, increase the fertilization potential of the oocyte.

  16. Failed fertilization during an in vitro fertilization cycle after oral ingestion of amantadine hydrochloride.

    PubMed

    Cowan, B D; Lucas, J A; Sopelak, V M; Lockard, V

    1988-10-01

    Oocyte fertilization occurred during three in vitro fertilization/embryo transfer (IVF/ET) treatment cycles of an infertile couple but failed during an IVF cycle when the husband took amantadine for prophylaxis against viral infection. Semen parameters were similar to those of other cycles attempted by this couple as well as to those of other IVF couples treated concurrently. We circumstantially suggest that amantadine, and potentially other ingestible medications or foods, while not spermicidal, may impair gamete function during IVF/ET.

  17. Failure of fertilization in in vitro fertilization: the "occult" male factor.

    PubMed

    Oehninger, S; Acosta, A A; Kruger, T; Veeck, L L; Flood, J; Jones, H W

    1988-08-01

    Failure of fertilization in patients undergoing in vitro fertilization (IVF) deserves extensive analysis for better prediction of the success or failure of this therapeutic modality. Consequently, we retrospectively studied the 52 couples in whom fertilization failed during Norfolk series 18 to 25, in an effort to establish the precise causes of failure. In the initial evaluation, pure oocyte abnormalities were identified in 19.2% of the cases; 32.6% showed sperm abnormalities, and a combination of oocyte and sperm anomalies was found in 7.7%. In 40.4% of the cases, failure of fertilization could not be explained. Reassessment of sperm morphology by new, strict criteria increased the identification of sperm abnormalities to 61.5% and of combined sperm and oocyte anomalies to 13.4%, for a total of 74.9% of sperm factors involved, as opposed to 40.3% in the original evaluation. The incidence of unexplained failed fertilization was substantially reduced, to 11.5%. In a control group (tubal infertility) matched by age, date, and stimulation, in whom fertilization occurred, 83.3% had normal sperm parameters as judged by the new criteria for morphology evaluation. This paper emphasizes the need for a more accurate diagnosis of sperm abnormalities to establish the true incidence of this factor in failed fertilization and to obtain information of prognostic value to patients and clinicians.

  18. In vitro fertilization and embryo transfer: a brief overview.

    PubMed Central

    DeCherney, A. H.

    1986-01-01

    The in vitro fertilization process breaks down into three essential components: induction of ovulation, fertilization of the oocyte, and development of embryos that are transferred into the uterus. Problems may arise resulting in failure at any one of these junctions. In 1984, the World Congress on In Vitro Fertilization was held, looking at 9,641 laparoscopies yielding 1,101 clinical pregnancies, with an overall pregnancy rate of 11 percent--clearly indicating that in vitro fertilization/embryo transfer (IVF/ET) was an idea whose time had come. Ovulation induction is monitored by both the use of ultrasound and daily estradiol levels, ultrasound indicating the number of oocytes that will be available for capture, and estradiol indicating in an indirect way the quality of those oocytes. It is a major aim in each patient to obtain at least four embryos, since this optimizes success rates. Ovulation induction at Yale is carried out with a high-dose human menopausal gonadotropin (HMG)/human chorionic gonadotropin (HCG) regimen. This regimen has insured us a success rate of 17 percent clinical pregnancies per laparoscopy. In the future, modification will occur in the process with cryopreservation of oocytes and embryos, and gamete manipulation. The modifications will be effected primarily to increase pregnancy rates. Research will continue mainly to delineate better biochemical markers for oocyte quality, but also to further explain the mystery of implantation. PMID:3532577

  19. [Programmed stimulation for in vitro fertilization].

    PubMed

    Imthurn, B; Macas, E; Hotz, E; Keller, P J

    1992-01-01

    A programmed stimulation protocol for in vitro fertilisation was evaluated in order to perform oocytes aspiration from Tuesday to Friday only. For this purpose, norethisterone acetate (Primolut-Nor) was given in a daily dose of 10 mg for 10-25 days, starting on the 2nd day of the preceding cycle and ending on a Monday. The stimulation was started on the following Friday by administering daily 0.1 mg D-Trp-6-LHRH s. c. (Decapeptyl). In group A (preliminary study) the treatment with HMG 150 IU/d (Pergonal) was initiated two days later. In group B the treatment with HMG was individualised with respect to the initial dose and the starting point. 10 ultrasonographic, needle-guided, transvaginal aspirations were performed from January to March 1990 in Group A, 29 aspirations from April to December 1990 in Group B. Indication for IVF in all patients was tubal infertility. All aspirations were possible from Tuesday to Friday. In Group A 5 of 15 cycles (33%) had to be cancelled. A total of 53 oocytes (5.3 +/- 2.3 per aspiration) were retrieved. Cleavage per oocyte took place in 62%. There was no pregnancy. In Group B, the cancellation rate could be reduced to 3% (1/30 cycles; p less than 0.025). 227 oocytes were recovered (7.8 +/- 3.1 per aspiration; p less than 0.05) and there was a higher cleavage rate (74%, n.s.). The clinical pregnancy rate (with allowance for progressive introduction of freezing) increased to 30% per stimulation cycle (9/30, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

  20. In Vitro Fertilization and Multiple Pregnancies

    PubMed Central

    2006-01-01

    increased need for medical and social support. The Technology Being Reviewed IVF was first developed as a method to overcome bilateral Fallopian tube obstruction. The procedure includes several steps: (1) the woman’s egg is retrieved from the ovaries; (2) exposed to sperm outside the body and fertilized; (3) the embryo(s) is cultured for 3 to 5 days; and (4) is transferred back to the uterus. IFV is considered to be one of the most effective treatments for infertility today. According to data from the Canadian Assisted Reproductive Technology Registry, the average live birth rate after IVF in Canada is around 30%, but there is considerable variation in the age of the mother and primary cause of infertility. An important advantage of IVF is that it allows for the control of the number of embryos transferred. An elective single embryo transfer in IVF cycles adopted in many European countries was shown to significantly reduce the risk of multiple pregnancies while maintaining acceptable birth rates. However, when number of embryos transferred is not limited, the rate of IVF-associated multiple pregnancies is similar to that of other treatments involving ovarian stimulation. The practice of multiple embryo transfer in IVF is often the result of pressures to increase success rates due to the high costs of the procedure. The average rate of multiple pregnancies resulting from IVF in Canada is currently around 30%. An alternative to IVF is IUI. In spite of reported lower success rates of IUI (pregnancy rates per cycle range from 8.7% to 17.1%) it is generally attempted before IVF due to its lower invasiveness and cost. Two major drawbacks of IUI are that it cannot be used in cases of bilateral tubal obstruction and it does not allow much control over the risk of multiple pregnancies compared with IVF. The rate of multiple pregnancies after IUI with COS is estimated to be about 21% to 29%. Ontario Health Insurance Plan Coverage Currently, the Ontario Health Insurance Plan covers

  1. Influence of L-arginine during bovine in vitro fertilization.

    PubMed

    Silva, Thiago Velasco Guimarães; da Silva, Bruno Baraúna; de Sá, André Luiz Alves; da Costa, Nathalia Nogueira; Sampaio, Rafael Vilar; Cordeiro, Marcela da Silva; Santana, Priscila Di Paula Bessa; Adona, Paulo Roberto; Santos, Simone do Socorro Damasceno; Miranda, Moysés dos Santos; Ohashi, Otávio Mitio

    2014-12-01

    The objective of this work was to evaluate the effect of using L-arginine during in vitro fertilization (IVF) on in vitro embryonic development using Bos taurus and Bos indicus semen. Effect of different concentrations (0, 1, 10 and 50 mM) of L-arginine, added to the IVF medium, was evaluated on the fertilization rate at 18 h post-fertilization (hpf), NO3(-)/NO2(-) production during IVF by the Griess colorimetric method (30 hpf), cleavage and blastocyst rates (on Day 2 and Day 7 of culture, respectively) and total blastocyst cell number (Day 7 of culture). The results reveal that the addition of 50 mM L-arginine to IVF medium, with either Bos taurus or Bos indicus spermatozoa, decreased the cleavage rate and blastocyst rate compared to the control group. Other concentrations did not affect embryo production. However, 1 mM L-arginine with Bos indicus semen increased the proportion of hatched blastocysts. These results indicate that high L-arginine concentrations may exhibit toxic effects on bovine gametes during in vitro fertilization. PMID:25651608

  2. Influence of L-arginine during bovine in vitro fertilization.

    PubMed

    Silva, Thiago Velasco Guimarães; da Silva, Bruno Baraúna; de Sá, André Luiz Alves; da Costa, Nathalia Nogueira; Sampaio, Rafael Vilar; Cordeiro, Marcela da Silva; Santana, Priscila Di Paula Bessa; Adona, Paulo Roberto; Santos, Simone do Socorro Damasceno; Miranda, Moysés dos Santos; Ohashi, Otávio Mitio

    2014-12-01

    The objective of this work was to evaluate the effect of using L-arginine during in vitro fertilization (IVF) on in vitro embryonic development using Bos taurus and Bos indicus semen. Effect of different concentrations (0, 1, 10 and 50 mM) of L-arginine, added to the IVF medium, was evaluated on the fertilization rate at 18 h post-fertilization (hpf), NO3(-)/NO2(-) production during IVF by the Griess colorimetric method (30 hpf), cleavage and blastocyst rates (on Day 2 and Day 7 of culture, respectively) and total blastocyst cell number (Day 7 of culture). The results reveal that the addition of 50 mM L-arginine to IVF medium, with either Bos taurus or Bos indicus spermatozoa, decreased the cleavage rate and blastocyst rate compared to the control group. Other concentrations did not affect embryo production. However, 1 mM L-arginine with Bos indicus semen increased the proportion of hatched blastocysts. These results indicate that high L-arginine concentrations may exhibit toxic effects on bovine gametes during in vitro fertilization.

  3. The relationship of semen parameters to fertilization in patients participating in a program of in vitro fertilization.

    PubMed

    Alper, M M; Lee, G S; Seibel, M M; Smith, D; Oskowitz, S P; Ransil, B J; Taymor, M L

    1985-12-01

    Approximately 80% of the patients in a program of in vitro fertilization (IVF) will fertilize an oocyte. The purpose of this study was to determine which parameters of the semen analysis influence fertilization in vitro. Of 120 patients participating in an in vitro fertilization program, 98 achieved fertilization of at least one mature oocyte and 22 did not. Ovulation induction was standardized and patients whose sperm was exposed to at least one mature oocyte (by light microscopy) were included in the study. Semen washing was accomplished using a "swim-up" technique. Semen parameters were assessed both before (raw) and after washing. Following insemination with 100,000 motile sperm, fertilization was determined by the presence of pronuclei or cleavage. Mean sperm count and motility were higher in patients who fertilized. However, morphology was similar. Fertilization was more likely to occur with a raw density greater than 104 million/ml and a motility greater than 64%, as well as with a density greater than 18 million/ml and a motility greater than 86% following washing. Furthermore, washing lowered sperm counts by 75% and increased motility by 25% but had no effect on morphology. This study demonstrates that sperm count and motility, but not morphology, influence fertilization in a program of in vitro fertilization and that patients with higher counts and motility have a greater probability of fertilization.

  4. Male-factor infertility and in vitro fertilization.

    PubMed

    Yates, C A; De Kretser, D M

    1987-06-01

    In vitro fertilization (IVF) was developed primarily as a treatment for female and idiopathic infertility. However, with the discovery that relatively few sperm are required to achieve fertilization in vitro, it was proposed that IVF could be used also as an effective treatment for male-factor infertility. This review deals with the work that has been carried out by various groups in this area of male-factor infertility. As the standards of classification and the presentation of results vary from group to group, this also shows that there is a need for some standardization of how patient selection and the presentation of results are carried out in the area of male-factor infertility.

  5. Purification of water for in vitro fertilization and embryo transfer.

    PubMed

    Fleetham, J; Mahadevan, M M

    1988-06-01

    The purpose of this study was to determine whether water obtained from the Milli-Q water purification system (Millipore, Mississauga, Canada) needed further purification for use in in vitro fertilization and embryo transfer. We describe a method for maintenance of the Milli-Q system. To assess water quality, alternate batches of culture media were prepared by using either Milli-Q water or Milli-Q water further treated by twice glass distillation. The percentage of mouse two-cell embryos that developed to blastocysts and the human in vitro fertilization-embryo transfer pregnancy rates were recorded for each batch. There were no significant differences in the parameters examined, indicating that further purification by twice glass distillation is not necessary if the Milli-Q system has been maintained as outlined.

  6. In vitro fertilization and embryo transfer in the rhesus monkey.

    PubMed

    Wolf, D P; Vandevoort, C A; Meyer-Haas, G R; Zelinski-Wooten, M B; Hess, D L; Baughman, W L; Stouffer, R L

    1989-08-01

    Twenty-three rhesus monkeys were subjected to 9 days of ovarian hyperstimulation with sequential exposure to human follicle-stimulating hormone (hFSH) and then human luteinizing hormone (hLH) + hFSH. Six animals (26%) did not exhibit sustained, elevated levels of circulating estradiol, primarily due to the occurrence of a premature surge of endogenous LH (n = 4). Seventeen animals (74%) responded with supraphysiologic levels of circulating estradiol (peak value: means = 4480 pg/ml) and received human chorionic gonadotropin (hCG) on Day 10. Oocytes were collected 26 h later by aspiration of large antral follicles. Oocyte quantity (means = 18/animal) and quality (63% mature) were evaluated by in vitro fertilization (IVF), embryonic development, and embryo transfer to foster mothers. Modified conditions for the successful fertilization of oocytes used a Tyrode's augmented (TALP) medium supplemented with 0.3% bovine serum albumin (BSA). Oocytes were inseminated at the metaphase II stage with ejaculated, washed sperm (50 100 x 10(3)/ml) preexposed at ambient temperature to caffeine and dibutyryl cyclic adenosine 3'5'-monophosphate. Successful fertilization ranged from 26% to 75%. In one experiment, 5 of 11 embryos produced by IVF developed in vitro to hatched blastocysts. Embryo freezing employed a propanediol-based protocol and was applied to early cleavage-stage embryos with 100% (5 of 5) post-thaw survival. Two frozen-thawed embryos were transferred transtubally on 3 occasions into rhesus monkeys during the early luteal phase of spontaneous menstrual cycles. One pregnancy resulted, which proceeded normally to the unassisted delivery of a male offspring 170 days after the LH surge. We conclude that this sequential regimen of human gonadotropins provides a cohort of oocytes from rhesus monkeys that will complete meiotic maturation and fertilize in vitro, with embryonic development proceeding in vitro and in vivo. The production of putative antibodies to human

  7. Microrobots for in vitro fertilization applications.

    PubMed

    Boukallel, M; Gauthier, M; Piat, E; Abadie, J; Roux, C

    2004-05-01

    The Micromanipulation and Micro-actuation Research Group at the LAB has activities related to biological and surgical applications. Concerning cells micromanipulation, our laboratory works in collaboration with the research team "Genetic and Reproduction" of the Besançon's hospital (France). The global final objective is the development of an automatic intra cytoplasmic sperm injection (ICSI) device in order to improve performances and ergonomics of current devices. In the future this new device will contain various modules: module for removal of cumulus cells, modules for characterization of oocytes, microinjection module, cells transport system. The first subsystem developed is a new single cell transport system. It consists in a so-called micropusher which pushes single cells without having contact with the external environment. This micropusher is a ferromagnetic particle (from 400 x 400 x 20 microm3 to 100 x 100 x 5 microm3) which follows the movement of a permanent magnet located under the biological medium. A 2D micro-positioning table moves this magnet under the glass slide. The pusher and cells positions are measured through an optical microscope with a CCD camera located above the biological medium. The second subsystem is developed to measure oocytes mechanical stiffness in order to sort them. We have then developed a micro/nano-force sensor based on the diamagnetic levitation principle: a glass tip end-effector (with 20 microm in diameter) is fixed on the equipment which is in levitation (0.5 mm in diameter, 100 mm in length). When a force is applied to the levitated glass tip, it moves to a new equilibrium position. Thanks to themeasurement of this displacement, the applied force can be measured. Since there is no contact and friction between the levitated tip and the fixed part, the resolution of this sensor is very high (10 nN).

  8. Risks of In Vitro Fertilization (IVF)

    MedlinePlus

    ... method of assisted reproduction in which a man’s sperm and a woman’s eggs are combined outside of ... remains under debate and study. However, when intracytoplasmic sperm injection (ICSI) is done along with IVF, there ...

  9. Fertilization capacity of cryopreserved Iberian ibex epididymal sperm in a heterologous in vitro fertilization assay.

    PubMed

    López-Saucedo, J; Santiago-Moreno, J; Fierro, R; Izquierdo, D; Coloma, M A; Catalá, M G; Jiménez, I; Paramio, M T

    2015-02-01

    In vitro fertilization (IVF) can be used to assess the fertilization capacity of sperm. Heterologous IVF may be useful when assessing that of wild animals as it is often difficult to obtain adequate numbers of naturally corresponding oocytes. The aim of the present study was to assess the fertilization capacity of frozen-thawed ibex epididymal spermatozoa via heterologous IVF involving the oocytes of prepubertal domestic goats. The effect on fertilization and embryo development of adding oestrous sheep serum (ESS) to the fertilization medium was also examined. Cumulus-oocyte complexes (COCs) were matured in TCM-199 for 24-27 h at 38.5°C in a 5% CO2 in air atmosphere. Frozen-thawed epididymal spermatozoa were selected by density gradient centrifugation. After maturation, the oocytes were co-incubated with spermatozoa in synthetic oviductal fluid (SOF) with different concentrations of ESS: SOF-C (0%), SOF-2 (2%) and SOF-20 (20%). At 17 h post-insemination (hpi), zygotes with one female and one male pronucleus (2PN) were categorised as normal; zygotes with 3PN were recorded as polyspermic, and oocytes with 1PN as asynchronous. Cleavage and blastocyst development were assessed at 48 and 168 hpi respectively. The percentage of zygotes with 2PN was higher in the SOF-2 than in the SOF-20 treatment group (27.7% versus 2.9% P < 0.05). The percentage of blastocysts formed with the SOF-C, SOF-2 and SOF-20 treatments were 1.1%, 7.5% and 0% respectively. These results show that the presence of 2% ESS achieves better results than the use of no serum or the standard 20% concentration. Heterologous IVF may be an effective method for predicting the fertilization capacity of ibex spermatozoa, and therefore perhaps that of other wild mountain ungulates. PMID:24286139

  10. Fertilization capacity of cryopreserved Iberian ibex epididymal sperm in a heterologous in vitro fertilization assay.

    PubMed

    López-Saucedo, J; Santiago-Moreno, J; Fierro, R; Izquierdo, D; Coloma, M A; Catalá, M G; Jiménez, I; Paramio, M T

    2015-02-01

    In vitro fertilization (IVF) can be used to assess the fertilization capacity of sperm. Heterologous IVF may be useful when assessing that of wild animals as it is often difficult to obtain adequate numbers of naturally corresponding oocytes. The aim of the present study was to assess the fertilization capacity of frozen-thawed ibex epididymal spermatozoa via heterologous IVF involving the oocytes of prepubertal domestic goats. The effect on fertilization and embryo development of adding oestrous sheep serum (ESS) to the fertilization medium was also examined. Cumulus-oocyte complexes (COCs) were matured in TCM-199 for 24-27 h at 38.5°C in a 5% CO2 in air atmosphere. Frozen-thawed epididymal spermatozoa were selected by density gradient centrifugation. After maturation, the oocytes were co-incubated with spermatozoa in synthetic oviductal fluid (SOF) with different concentrations of ESS: SOF-C (0%), SOF-2 (2%) and SOF-20 (20%). At 17 h post-insemination (hpi), zygotes with one female and one male pronucleus (2PN) were categorised as normal; zygotes with 3PN were recorded as polyspermic, and oocytes with 1PN as asynchronous. Cleavage and blastocyst development were assessed at 48 and 168 hpi respectively. The percentage of zygotes with 2PN was higher in the SOF-2 than in the SOF-20 treatment group (27.7% versus 2.9% P < 0.05). The percentage of blastocysts formed with the SOF-C, SOF-2 and SOF-20 treatments were 1.1%, 7.5% and 0% respectively. These results show that the presence of 2% ESS achieves better results than the use of no serum or the standard 20% concentration. Heterologous IVF may be an effective method for predicting the fertilization capacity of ibex spermatozoa, and therefore perhaps that of other wild mountain ungulates.

  11. Monozygous double inner cell masses in mouse blastocysts following fertilization in vitro and in vivo.

    PubMed

    Chida, S

    1990-06-01

    The incidence of human identical twins after in vitro fertilization and embryo transfer (IVF-ET) is estimated to be higher than the commonly accepted incidence after conception in vivo. This paper reports that 8 mouse blastocysts which had double inner cell masses in 261 blastocysts (3.1%) were identified after fertilization in vitro and they developed into trophoblastic outgrowth formation with two inner cell masses. In contrast, only 3 monozygous double inner cell masses were observed in 526 blastocysts (0.6%) which were fertilized in vivo and cultured from the two-cell stage in vitro. It is therefore possible that fertilization in vitro predisposes to monozygous twinning.

  12. In vitro and in vivo fertility of ram semen cryopreserved in different extenders.

    PubMed

    Valente, S S; Pereira, R M; Baptista, M C; Marques, C C; Vasques, M I; Pereira, M V C Silva; Horta, A E M; Barbas, J P

    2010-01-01

    Seminal traits of frozen-thawed (FT) ram semen and in vitro and field fertility in native Portuguese breeds were evaluated in 4 experiments. In exp. 1 and 2 the cryopreservation capacity of 2 extenders, E1 (15% egg yolk-EY) and E2 (4.5% EY and trehalose) was compared through morphological evaluation and in vitro fertilizability of FT ram semen. Exp. 3 aimed to determine the usefulness of in vitro homologous/heterologous fertilization tests as tools for predicting ram fertility. Exp. 4 was conducted to verify if the identified differences between the 2 extenders could be confirmed by field fertility. E1 showed a better cryoprotective action expressed by higher in vitro and field fertility results. In conclusion, EY is difficult to be replaced in ram semen extenders. Heterologous fertilization seems to be a useful tool for predicting fertility of FT ram semen.

  13. Low-cost in vitro fertilization: current insights.

    PubMed

    Teoh, Pek Joo; Maheshwari, Abha

    2014-01-01

    Despite the development of in vitro fertilization (IVF) more than 30 years ago, the cost of treatment remains high. Furthermore, over the years, more sophisticated technologies and expensive medications have been introduced, making IVF increasingly inaccessible despite the increasing need. Globally, the option to undergo IVF is only available to a privileged few. In recent years, there has been growing interest in exploring strategies to reduce the cost of IVF treatment, which would allow the service to be provided in low-resource settings. In this review, we explore the various ways in which the cost of this treatment can be reduced.

  14. Low-cost in vitro fertilization: current insights

    PubMed Central

    Teoh, Pek Joo; Maheshwari, Abha

    2014-01-01

    Despite the development of in vitro fertilization (IVF) more than 30 years ago, the cost of treatment remains high. Furthermore, over the years, more sophisticated technologies and expensive medications have been introduced, making IVF increasingly inaccessible despite the increasing need. Globally, the option to undergo IVF is only available to a privileged few. In recent years, there has been growing interest in exploring strategies to reduce the cost of IVF treatment, which would allow the service to be provided in low-resource settings. In this review, we explore the various ways in which the cost of this treatment can be reduced. PMID:25187741

  15. Hair mercury concentrations and in vitro fertilization (IVF) outcomes among women from a fertility clinic.

    PubMed

    Wright, Diane L; Afeiche, Myriam C; Ehrlich, Shelley; Smith, Kristen; Williams, Paige L; Chavarro, Jorge E; Batsis, Maria; Toth, Thomas L; Hauser, Russ

    2015-01-01

    Total hair mercury (Hg) was measured among 205 women undergoing in vitro fertilization (IVF) treatment and the association with prospectively collected IVF outcomes (229 IVF cycles) was evaluated. Hair Hg levels (median=0.62ppm, range: 0.03-5.66ppm) correlated with fish intake (r=0.59), and exceeded the recommended EPA reference of 1ppm in 33% of women. Generalized linear mixed models with random intercepts accounting for within-woman correlations across treatment cycles were used to evaluate the association of hair Hg with IVF outcomes adjusted for age, body mass index, race, smoking status, infertility diagnosis, and protocol type. Hair Hg levels were not related to ovarian stimulation outcomes (peak estradiol levels, total and mature oocyte yields) or to fertilization rate, embryo quality, clinical pregnancy rate or live birth rate.

  16. Maturation, fertilization, and development of marmoset monkey oocytes in vitro.

    PubMed

    Gilchrist, R B; Nayudu, P L; Hodges, J K

    1997-01-01

    This study was conducted to investigate 1) the capacity of in vitro-matured (IVM) marmoset oocytes to be fertilized and to support embryonic development in vitro and 2) oocyte meiotic maturation in relation to in vivo FSH administration, follicle size, and oocyte-cumulus cell status. Pairs of ovaries were collected on Day 4 of the follicular phase from adult females receiving either 1) human FSH (3 IU; n = 5) or 2) control (saline; n = 5) daily for 4 days. Antral follicles were excised from ovaries and separated into classes according to size: class 1 (660-840 microm), class 2 (> 840-1000 microm), class 3 (> 1000-1400 microm), and class 4 (> 1400 microm). A total of 823 partially naked and cumulus-enclosed oocytes (CEOs) were released from follicles and cultured in vitro. Cumulus cells remaining after 22 h were removed, metaphase II (MII) oocytes were inseminated with epididymal sperm, and resulting embryos were cultured until developmental arrest. Fluorescence microscopy was used to assess oocyte meiotic and embryo developmental progression. Oocyte germinal vesicle breakdown (GVB)- and MII-competencies increased significantly with follicular size (p < 0.01 and p < 0.0001, respectively), although they were independent of oocyte-cumulus cell associations. After 24 and 32 h in vitro, 69% and 93%, respectively, of CEOs with MII competence had completed meiotic maturation, and the rate of nuclear maturation increased progressively with follicle size (p < 0.01) and with the association of cumulus cells (p < 0.01). In vivo FSH priming slightly improved oocyte GVB- and MII-competencies (p < 0.01 and p < 0.05, respectively) and decreased the time required to achieve MII (p < 0.01). IVM oocytes from all follicle sizes fertilized (78-92%) in vitro, with 27% developing to morula- and 4% to blastocyst-stage embryos. This study demonstrates for the first time that IVM New World primate oocytes are able to support advanced preimplantation embryonic development in vitro. Oocyte

  17. Microdroplet in vitro fertilization can reduce the number of spermatozoa necessary for fertilizing oocytes.

    PubMed

    Hasegawa, Ayumi; Mochida, Keiji; Tomishima, Toshiko; Inoue, Kimiko; Ogura, Atsuo

    2014-01-01

    Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is mostly performed using about 80-500 µl drops, it is expected that the number of spermatozoa used for insemination can be reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using frozen-thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates were low (13-15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number of inseminated spermatozoa per oocyte can be reduced to 1/96-1/240 by this method. In two separate embryo transfer experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic reasons.

  18. The association of in vitro fertilization and perinatal morbidity.

    PubMed

    Kalra, Suleena Kansal; Molinaro, Thomas A

    2008-09-01

    In recent years, there has been increasing concern regarding the safety of in vitro fertilization (IVF) because of the potential health impact on these infants. Multiple pregnancy contributes the vast majority of morbidity associated with IVF and, initially, many thought that adverse outcomes after IVF were solely attributable to the high incidence of twin pregnancies. More recently, multiple studies have suggested that IVF singleton pregnancies may be at increased risk for preterm birth, low birth weight, congenital anomalies, perinatal mortality, and several other pregnancy-related complications compared with unassisted singleton pregnancies. We have focused this review on the increased risk of adverse outcomes in IVF singleton conceptions compared with that of unassisted conceptions. The available evidence evaluating the association between IVF and low birth weight, preterm delivery, placental abruption, preeclampsia, congenital anomalies, and perinatal mortality in singleton pregnancies is summarized. In addition, data reporting an increased risk of congenital and chromosomal anomalies after IVF are presented.

  19. Sperm midpiece apoptotic markers: impact on fertilizing potential in in vitro fertilization and intracytoplasmic sperm injection.

    PubMed

    Talarczyk-Desole, Joanna; Kotwicka, Małgorzata; Jendraszak, Magdalena; Pawelczyk, Leszek; Murawski, Marek; Jędrzejczak, Piotr

    2016-04-01

    The aim of this study was to investigate the relationship between apoptotic markers present in human spermatozoa, namely phosphatidylserine translocation (PST) from the inner to the outer layer of the cytomembrane and the active form of caspase-3 (c3) versus the fertilizing potential of male gametes in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) models. A total of 116 male patients treated with their partners for infertility underwent basic semen analysis and an assessment of the presence of PST and the active c3 in sperm using flow cytometry. Forty patients underwent IVF, group A, while 76 patients underwent ICSI, group B. The fertilizing potential of the gametes was measured as the percentage of oocytes with pronuclei present after either procedure. PST and active c3 were identified in vital gametes, mainly in the midpiece area. Concentration, motility, morphology, and viability of spermatozoa strongly negatively correlated with both markers. In group A, a negative correlation between both markers and the success rate of conventional IVF was observed (r = -0.4, p = 0.04 for PST; r = -0.4, p = 0.02 for active c3, respectively). In group B, the success rate of ICSI did not correlate with either marker (r = -0.2, p = 0.85 for PST and r = 0.1, p = 0.51 for active c3). The two apoptotic markers localized in the sperm midpiece area may affect their function not only by decreasing basic andrologic parameters but also by reducing the probability of conception. Therefore, analysis of PST and active c3 in the sperm of patients undergoing infertility treatment should be recommended.

  20. In vitro fertilization/intracytoplasmic sperm injection for male infertility

    PubMed Central

    Merchant, Rubina; Gandhi, Goral; Allahbadia, Gautam N.

    2011-01-01

    Progress in the field of assisted reproduction, and particularly micromanipulation, now heralds a new era in the management of severe male factor infertility, not amenable to medical or surgical correction. By overcoming natural barriers to conception, in vitro fertilization and embryo transfer (IVF-ET), subzonal sperm insemination, partial zona dissection, and intracytoplasmatic injection of sperm (ICSI) now offer couples considered irreversibly infertile, the option of parenting a genetically related child. However, unlike IVF, which necessitates an optimal sperm number and function to successfully complete the sequence of events leading to fertilization, micromanipulation techniques, such as ICSI, involving the direct injection of a spermatozoon into the oocyte, obviate all these requirements and may be used to alleviate severe male factor infertility due to the lack of sperm in the ejaculate due to severely impaired spermatogenesis (non-obstructive azoospermia) or non-reconstructable reproductive tract obstruction (obstructive azoospermia). ICSI may be performed with fresh or cryopreserved ejaculate sperm where available, microsurgically extracted epididymal or testicular sperm with satisfactory fertilization, clinical pregnancy, and ongoing pregnancy rates. However, despite a lack of consensus regarding the genetic implications of ICSI or the application and efficacy of preimplantation genetic diagnosis prior to assisted reproductive technology (ART), the widespread use of ICSI, increasing evidence of the involvement of genetic factors in male infertility and the potential risk of transmission of genetic disorders to the offspring, generate major concerns with regard to the safety of the technique, necessitating a thorough genetic evaluation of the couple, classification of infertility and adequate counseling of the implications and associated risks prior to embarking on the procedure. The objective of this review is to highlight the indications, advantages

  1. The effect of endometriomas on in vitro fertilization outcome.

    PubMed

    Dlugi, A M; Loy, R A; Dieterle, S; Bayer, S R; Seibel, M M

    1989-12-01

    To determine the effect of ovarian endometriomas on in vitro fertilization (IVF) outcome, two groups of patients were studied. Group I consisted of seven patients with ovarian endometriomas and severe pelvic adhesions treated for a total of 12 cycles. Group II patients consisted of eight patients with hydrosalpinges and comparable pelvic adhesions treated for a total of 27 cycles. There were no differences in the number of days required for stimulation or in the serum estradiol levels attained between the two groups. Group I patients were noted to have significantly fewer preovulatory follicles (1.42 vs 3.33, P less than 0.005), cycles with fertilization (28 vs 84%, P less than 0.005), and embryos transferred (0.78 vs 2.56, P = 0.01) than Group II patients. Three pregnancies occurred in Group II, while there were no conceptions among Group I patients. This study suggests that the presence of an ovarian endometrioma(s) has an adverse effect on IVF outcome and suggests that patients with ovarian endometriomas should have them removed prior to undergoing IVF.

  2. In vitro fertilization and the right to reproduce.

    PubMed

    Uniacke, Suzanne

    1987-07-01

    The assumption that people have a right to have children is considered in relation to the question of whether society has an obligation to allocate resources to make clinical in vitro fertilization available, at least to married couples where the woman is infertile and the couple provides the ovum and sperm. The author analyzes various issues that need to be addressed: the distress that accompanies infertility versus the fact that infertility is not a life- or health-threatening condition; the distinction between basic needs or desires on the one hand and claim rights on the other; the extent of the obligations that affluent societies have to fund other services related to the bearing and rearing of children; the costs and benefits of relieving infertility through IVF versus other means; and the concept of compensation to one who has been injured or harmed unjustly, here by the involuntary loss of reproductive capacity.

  3. Mother-daughter in vitro fertilization triplet surrogate pregnancy.

    PubMed

    Michelow, M C; Bernstein, J; Jacobson, M J; McLoughlin, J L; Rubenstein, D; Hacking, A I; Preddy, S; Van der Wat, I J

    1988-02-01

    A successful triplet pregnancy has been established in a surrogate gestational mother following the transfer of five embryos fertilized in vitro. The oocytes were donated by her biological daughter, and the sperm obtained from the daughter's husband. The daughter's infertility followed a total abdominal hysterectomy performed for a postpartum hemorrhage as a result of a placenta accreta. Synchronization of both their menstrual cycles was obtained using oral contraceptive suppression for 2 months, followed by stimulation of both the surrogate gestational mother and her daughter such that embryo transfer would occur at least 48 hr after the surrogate gestational mother's own ovulation. This case raises a number of medical, social, psychological, and ethical issues. PMID:3367072

  4. Bovine in vitro fertilization: in vitro oocyte maturation and sperm capacitation with heparin.

    PubMed

    Parrish, John J

    2014-01-01

    As a result of research in the 1980s on in vitro maturation, sperm capacitation, and in vitro fertilization, the bovine is now one of the important models for development. Further, the current production of bovine embryos in vitro rivals that of in vivo embryo production for commercial applications. Researchers of today may be unaware of why decisions were made in the procedures. This review addresses the state of the art at the time of the work by Parrish et al. (Bovine in vitro fertilization with frozen thawed semen. Theriogenology 1986;25:591-600), and how later work would explain success or failure of competing procedures. Important was the use of frozen semen and heparin capacitation, because this allowed future researchers/practitioners to change sperm numbers and capacitation conditions to adjust for variations among bulls. The large numbers of citation of the original work stand the testament of time in the repeatability and success of the procedures. The work was done within the environment of the N.L. First laboratory and the unique interactions with a large number of talented graduate students, postdoctoral researchers, and technicians.

  5. Psychological aspects of in vitro fertilization: a review.

    PubMed

    Eugster, A; Vingerhoets, A J

    1999-03-01

    This paper reviews psychological research within the context of in vitro fertilization (IVF). The focus will be on psychological reactions before entering an IVF-procedure, during an IVF-treatment, and after both unsuccessful and successful IVF. The effects of psychosocial factors on the treatment outcome after IVF and interventions on conception rates will also be discussed. Undergoing an IVF-treatment is an emotional and physical burden, for both the woman and her partner. Research results suggest that couples entering an IVF-treatment program are, in general, psychologically well adjusted. Concerning reactions during the treatment, both women and men experience waiting for the outcome of the IVF-treatment and an unsuccessful IVF. as most stressful. Common reactions during IVF are anxiety and depression, while after an unsuccessful IVF, feelings of sadness, depression and anger prevail. After a successful IVF-treatment, IVF-parents experience more stress during pregnancy than 'normal fertile' parents. Mothers with children conceived by IVF express a higher quality of parent child relationship than mothers with a naturally conceived child. Research further suggests that psychosocial factors, like ineffective coping strategies, anxiety and/or depression are associated with a lower pregnancy rate following IVF-procedures. In addition, support has been found suggesting that stress reduction through relaxation training or behavioral treatment improves conception rates.

  6. In vitro fertilization for male infertility: when and how?

    PubMed

    Hall, J; Fishel, S

    1997-12-01

    The first observation that in vitro fertilization (IVF) was useful for treating oligozoospermia and oligoasthenozoospermia was reported by Fishel and Edwards in 1982. This was followed by a series of cases indicating the value of IVF in such cases. Conventional IVF has been modified and refined to achieve increased rates of conception in cases of male factor infertility. Methods such as high insemination concentration IVF for the treatment of teratozoospermia and microscopic IVF for the treatment of oligozoospermia have had some impact on fertilization and pregnancy rates; however, reports of success are varied. The recent advent of micromanipulation and, in particular, intracytoplasmic sperm injection (ICSI) has overshadowed the use of these modified IVF procedures. Because of the high fertilization and pregnancy rates achieved with ICSI, other micromanipulation techniques (subzonal insemination and partial zona dissection) have been abandoned; there have also been suggestions that other more conventional techniques, i.e. IVF, should also be abandoned and that ICSI become the sole technique for the treatment of infertility. The rapid increase in the number of centres using ICSI has led to extreme pressure for individual units to achieve high fertilization and pregnancy rates and there is a temptation to assign all patients to ICSI treatment. It is important that, in this highly competitive environment, new techniques are not applied haphazardly and reduced to the mere injection of gametes and achievement of pregnancy regardless of the cause of infertility. In his 1986 IVF--Historical Perspective, Fishel quoted Auguste Comte: 'to understand science it is necessary to know its history'. IVF has much recent history in animal and also human work. Although ICSI is the most significant therapeutic advance in male infertility treatment, its application to human IVF is only 4 years old, with a paucity of animal studies on which to rely. For this reason IVF still plays a

  7. In vitro fertilization for male infertility: when and how?

    PubMed

    Hall, J; Fishel, S

    1997-12-01

    The first observation that in vitro fertilization (IVF) was useful for treating oligozoospermia and oligoasthenozoospermia was reported by Fishel and Edwards in 1982. This was followed by a series of cases indicating the value of IVF in such cases. Conventional IVF has been modified and refined to achieve increased rates of conception in cases of male factor infertility. Methods such as high insemination concentration IVF for the treatment of teratozoospermia and microscopic IVF for the treatment of oligozoospermia have had some impact on fertilization and pregnancy rates; however, reports of success are varied. The recent advent of micromanipulation and, in particular, intracytoplasmic sperm injection (ICSI) has overshadowed the use of these modified IVF procedures. Because of the high fertilization and pregnancy rates achieved with ICSI, other micromanipulation techniques (subzonal insemination and partial zona dissection) have been abandoned; there have also been suggestions that other more conventional techniques, i.e. IVF, should also be abandoned and that ICSI become the sole technique for the treatment of infertility. The rapid increase in the number of centres using ICSI has led to extreme pressure for individual units to achieve high fertilization and pregnancy rates and there is a temptation to assign all patients to ICSI treatment. It is important that, in this highly competitive environment, new techniques are not applied haphazardly and reduced to the mere injection of gametes and achievement of pregnancy regardless of the cause of infertility. In his 1986 IVF--Historical Perspective, Fishel quoted Auguste Comte: 'to understand science it is necessary to know its history'. IVF has much recent history in animal and also human work. Although ICSI is the most significant therapeutic advance in male infertility treatment, its application to human IVF is only 4 years old, with a paucity of animal studies on which to rely. For this reason IVF still plays a

  8. Simulated Microgravity Influences Bovine Oocyte In Vitro Fertilization and Preimplantation Embryo Development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study was to investigate whether in vitro fertilization and preimplantation embryos exposed to a simulated microgravity environment in vitro would improve, or be deleterious to, their fertilization and embryonic development. A Rotating Cell Culture System™ (RCCS) bioreactor with a Hi...

  9. [Application of microfluidics in sperm isolation and in vitro fertilization].

    PubMed

    Li, Fang-Fang; Wang, Xiao-Ying; Zhou, Shu-Min; You, Fan

    2014-05-01

    Due to the low effectiveness of traditional assisted reproductive technology (ART), new technological possibilities are constantly explored. Lots of studies have demonstrated the potential of microfluidics to revolutionize the fundamental processes of in vitro fertilization (IVF). With the advantages of high efficiency, short time, harmless collection, real-time observation of separation, similar microenvironment, and automation, the application of microfluidics in sperm isolation and IVF has shown an evident superiority over the conventional approaches and provided a new platform for ART. This review highlights the application of various microfluidic techniques in sperm motility assessment and isolation, sperm chemotaxis assay, IVF, sperm concentration, and sperm separation and enrichment in recent years. It also briefly introduces the basic principles, structural design, and operation processes of the microfluidic platform, focusing on the advantages and disadvantages of each method and the potential of their clinical application. Obviously, there are still some challenges to the application of microfluidics in ART. However, it is believed that the development of this new technology would be toward a highly integrated application of several steps in one single device, known as IVF-lab-on-a-chip.

  10. Abnormal chromosome behavior in human oocytes which remained unfertilized during human in vitro fertilization.

    PubMed

    Spielmann, H; Krüger, C; Stauber, M; Vogel, R

    1985-09-01

    Chromosomal abnormalities and abnormal embryonic development have previously been observed after human in vitro fertilization (IVF). Chromosomal abnormalities may arise not only after fertilization but even earlier during meiotic maturation of human oocytes in culture. Since chromosomal analysis is simple in oocytes during meiotic maturation, the chromosomal status was analyzed in oocytes which remained unfertilized in a human in vitro fertilization program. In 50 fertilization attempts the chromosomes of 62 unfertilized oocytes could be analyzed; 45 of them were in the process of meiotic maturation. In three oocytes two small polar bodies were observed 16-18 hr after insemination in the absence of fertilization. In one oocyte abnormal chromosome behavior was found during the first meiotic division, and in four oocytes during metaphase of the second meiotic division. These data suggest that chromosomal analysis of unfertilized oocytes in human IVF may improve the understanding human oocyte maturation and fertilization.

  11. Combined electroejaculation and in vitro fertilization in the evaluation and treatment of anejaculatory infertility.

    PubMed

    Randolph, J F; Ohl, D A; Bennett, C J; Ayers, J W; Menge, A C

    1990-02-01

    Seven couples underwent combined electroejaculation and in vitro fertilization for anejaculatory infertility after a failed regimen of electroejaculation and intrauterine insemination. Two pregnancies resulted, one proceeding to a term vaginal delivery and one ending in a 6-week spontaneous abortion. Poor sperm binding and no fertilization were seen in two of the couples. Fertilization of the oocytes but no subsequent pregnancy were seen in the other three couples. Combined in vitro fertilization and gamete intrafallopian transfer was performed in four of the couples. The combination of electroejaculation and in vitro fertilization offers the opportunity to evaluate the female pelvis, observe the sperm-oocyte interaction, and achieve a pregnancy in couples with anejaculatory infertility.

  12. Transcapillary fluid dynamics during ovarian stimulation for in vitro fertilization.

    PubMed

    Tollan, A; Holst, N; Forsdahl, F; Fadnes, H O; Oian, P; Maltau, J M

    1990-02-01

    Transcapillary fluid dynamics were studied in 10 women during ovarian stimulation for in vitro fertilization. The examinations were done on the first day of stimulation (day 3 of the menstrual cycle, mean serum estradiol concentration 0.2 nmol/L), and the day before oocyte aspiration (day 10 to 12, mean serum estradiol concentration 6.8 nmol/L). Interstitial colloid osmotic pressure was measured on the thorax at heart level by the "wick" method, and interstitial hydrostatic pressure by the "wick-in-needle" method. Plasma colloid osmotic pressure decreased (mean, 2.0 mm Hg; p less than 0.002) and interstitial colloid osmotic pressure increased (mean, 1.0 mm Hg; p less than 0.02) during hormonal stimulation. This implies a reduced transcapillary colloid osmotic gradient (plasma colloid osmotic pressure--interstitial colloid osmotic pressure), probably because of increased capillary permeability to plasma proteins. Hemoglobin and hematocrit were significantly reduced, and body weight and foot volume significantly increased. These results demonstrate that during ovarian stimulation there are both water retention and augmented filtration of fluid from the vascular to the interstitial compartment. This may be of significance for the pathophysiologic condition in the ovarian hyperstimulation syndrome.

  13. Zebrafish Reproduction: Revisiting In Vitro Fertilization to Increase Sperm Cryopreservation Success

    PubMed Central

    Hagedorn, Mary; Carter, Virginia L.

    2011-01-01

    Although conventional cryopreservation is a proven method for long-term, safe storage of genetic material, protocols used by the zebrafish community are not standardized and yield inconsistent results, thereby putting the security of many genotypes in individual laboratories and stock centers at risk. An important challenge for a successful zebrafish sperm cryopreservation program is the large variability in the post-thaw in vitro fertilization success (0 to 80%). But how much of this variability was due to the reproductive traits of the in vitro fertilization process, and not due to the cryopreservation process? These experiments only assessed the in vitro process with fresh sperm, but yielded the basic metrics needed for successful in vitro fertilization using cryopreserved sperm, as well. We analyzed the reproductive traits for zebrafish males with a strict body condition range. It did not correlate with sperm volume, or motility (P>0.05), but it did correlate with sperm concentration. Younger males produced more concentrated sperm (P<0.05). To minimize the wastage of sperm during the in vitro fertilization process, 106 cells/ml was the minimum sperm concentration needed to achieve an in vitro fertilization success of ≥ 70%. During the in vitro process, pooling sperm did not reduce fertilization success (P>0.05), but pooling eggs reduced it by approximately 30 to 50% (P<0.05). This reduction in fertilization success was due not to the pooling of the females' eggs, but to the type of tools used to handle the eggs. Recommendations to enhance the in vitro process for zebrafish include: 1) using males of a body condition closer to 1.5 for maximal sperm concentration; 2) minimizing sperm wastage by using a working sperm concentration of 106 motile cells/ml for in vitro fertilization; and 3) never using metal or sharp-edged tools to handle eggs prior to fertilization. PMID:21698162

  14. Experience with intravaginal culture for in vitro fertilization (IVF).

    PubMed

    Costoya, A L; Cafatti, C M; Gadán, A A

    1991-12-01

    It has been reported that intravaginal culture (IVC) yields results comparable to those obtained with classic IVF techniques; furthermore, it can simplify and reduce the costs of the procedure. In our experience with IVC however, only 8 of 78 mature oocytes inseminated (10.2%) fertilized. Possible causes of this low fertilization rate were analyzed. In our opinion this new and attractive technique needs further evaluation before being extended to common clinical practice.

  15. Ovarian cyst aspiration prior to initiating ovarian hyperstimulation for in vitro fertilization.

    PubMed

    Silverberg, K M; Olive, D L; Schenken, R S

    1990-06-01

    Twenty-three consecutive patients presenting for in vitro fertilization were evaluated with transvaginal sonography on cycle day 3, prior to initiating ovarian hyperstimulation. Three of these patients were noted to have large ovarian cysts. All three underwent transvaginal aspiration of the cysts, followed immediately by initiation of ovarian hyperstimulation. Following oocyte retrieval, in vitro fertilization, and embryo transfer, all three women achieved pregnancy. We conclude that cyst aspiration is not contraindicated when an ovarian cyst is encountered in the follicular phase of an in vitro fertilization cycle.

  16. The fertilization ability and developmental competence of bovine oocytes grown in vitro.

    PubMed

    Makita, Miho; Ueda, Mayuko; Miyano, Takashi

    2016-08-25

    In vitro growth culture systems for oocytes are being developed in several mammalian species. In these growth culture systems, in vitro grown oocytes usually have lower blastocyst formation than in vivo grown oocytes after in vitro fertilization. Furthermore, there have been a few reports that investigated the fertilization ability of in vitro grown oocytes in large animals. The purpose of this study was to investigate the fertilization process and developmental competence of bovine oocytes grown in vitro. Oocyte-granulosa cell complexes collected from bovine early antral follicles (0.4-0.7 mm in diameter) were cultured for growth with 17β-estradiol and androstenedione for 14 days and matured in vitro. These oocytes were then inseminated for 6 or 12 h, and further cultured for development up to 8 days in vitro. After growth culture, oocytes grew from 95 µm to around 120 µm and acquired maturation competence (79%). Although fertilization rates of in vitro grown oocytes were low after 6 h of insemination, 34% of in vitro grown oocytes fertilized normally after 12 h of insemination, having two polar bodies and two pronuclei with a sperm tail, and 22% of these oocytes developed into blastocysts after 8 days of culture. The fertilization and blastocyst formation rates were similar to those of in vivo grown oocytes. In addition, blastocyst cell numbers were also similar between in vitro and in vivo grown oocytes. In conclusion, in vitro grown bovine oocytes are similar to in vivo grown oocytes in fertilization ability and can develop into blastocysts.

  17. [In vitro fertilization and systemic lupus erythematosus or antiphospholipid syndrome: An update].

    PubMed

    Orquevaux, P; Masseau, A; Le Guern, V; Gayet, V; Vauthier, D; Boutin, D; Wechsler, B; Morel, N; Guettrot-Imbert, G; Pennaforte, J-L; Piette, J-C; Costedoat-Chalumeau, N

    2015-03-01

    Fertility is not impaired in systemic lupus erythematosus or antiphospholipid syndrome, but, similarly to the general population, these patients may undergo in vitro fertilization. This type of treatment increases the risk of lupus flare, thrombosis, and ovarian hyperstimulation syndrome. This review will focus on in vitro fertilization in systemic lupus erythematosus or antiphospholipid syndrome. Literature data are relatively scant with only 3 reported studies. The first one included 17 patients and 63 cycles of induction ovulation/in vitro fertilization leading to 25 % of lupus flare, no thrombosis, and 3 % of ovarian hyperstimulation syndrome. The second study included 10 patients and 40 cycles of in vitro fertilization showing 31 % of lupus flare, no thrombosis and no ovarian hyperstimulation syndrome. The last one included 34 patients and 83 procedures of in vitro fertilization leading to 8 % of flares, 5 % of thrombosis and no ovarian hyperstimulation syndrome. Interestingly, in this last study, half of the complications were explained by poor adherence to treatment. These data are reassuring but it is important to remember that in vitro fertilization should be scheduled and carefully supervised in the same way as the high-risk pregnancies occurring in these patients.

  18. Preimplantation development following in vitro fertilization of mouse oocytes: effects of timing of superovulation and preincubation in vitro.

    PubMed

    Edgar, D H; Whalley, K M; Mills, J A

    1987-04-01

    The early embryonic development of in vitro fertilized oocytes was assessed following superovulation in F1 hybrid (C57BL/6 X CBA/Ca) mice. Decreasing the time interval between the administration of constant doses of pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) resulted in decreases in the frequency of development to the blastocyst stage but had no significant effect on development to the two-cell stage. Preincubation of postovulatory oocytes in vitro prior to insemination did not compensate for the reduced preovulatory development in vivo but resulted in decreases in the frequency of development to the blastocyst stage. The results indicate that inadequate preovulatory development of superovulated mouse oocytes can adversely affect the preimplantation development of in vitro fertilized embryos in the absence of a visible inhibitory effect on development to the two-cell stage and also that preincubation of postovulatory oocytes in vitro prior to fertilization reduces subsequent developmental capacity.

  19. [Poor Ovarian Response to Stimulation for In Vitro Fertilization].

    PubMed

    Spremović-Radjenović, Svetlana; Bila, Jovan; Gudović, Aleksandra; Vidaković, Snežana; Dokić, Milan; Radunović, Nebojša

    2015-01-01

    The term "poor respond (POR) patients" is used for the group of women who respond badly to usual doses of gonadotropins in in vitro fertilization (IVF) treatments; the consequence is low pregnancy rate. A consensus was reached on the minimal criteria needed to define POR. At least two of the following three features must be present: 1. advanced maternal age (40 years or more) 2. previous POR (3 or less oocytes with a conventional stimulation protocol) 3. abnormal ovarian reserve (AMH 0.5-1.1 ng/ml or AFC 5-7).The aim is to find better therapeutic options for these patients. Increased levels of day 3 follicle stimulating hormone (FSH) and estradiol (E2), as well as decreased levels of anti-Müllerian hormone (AMH) and antral follicle count (AFC), can be used to assess ovarian reserve, as indirect predictive tests. A larger number of well designed, large scale, randomized, controlled trials are needed to assess the efficacy of different management strategies for poor responders: flare up gonadotropin releasing hormone (GnRH) agonist protocols, modified long GnRH agonist mini-dose protocols, luteal initiation GnRH agonist stop protocol, pretreatment with estradiol--GnRH antagonist in luteal phase, natural cycle aspiration or natural cycle aspiration GnRH antagonist controlled, adjuvant therapy with growth hormone or dehydroepiandrosterone (DHEA). The results of up to now used protocols are unsatisfactory and stimulation of the ovulation in poor responders remains a challenge, especially when bearing in mind that in the majority of cases the patients will be menopausal in relatively short period of time.

  20. Heat stress during in vitro fertilization decreases fertilization success by disrupting anti-polyspermy systems of the oocytes.

    PubMed

    Sakatani, Miki; Yamanaka, Kenichi; Balboula, Ahmed Z; Takenouchi, Naoki; Takahashi, Masashi

    2015-01-01

    Low pregnancy rates during the summer are due, in part, to reduced fertilization. Given that elevated temperature is associated with this season, we investigated the effect of heat stress during fertilization using an in vitro model. Three experiments were performed to determine the mechanism by which exposure to elevated temperature disrupts fertilization. Oocytes were fertilized for 6 hr at 38.5°C or 41.0°C or 40.0°C with non-pre-incubated sperm, or for 6 hr at 38.5°C with sperm that had been pre-incubated at 38.5°C or 41.0°C for 4 hr. In each experiment, zygotes were cultured at 38.5°C in 5% CO(2) and 5% O(2). Rates of cleavage and blasocyst formation were reduced when fertilization occurs at elevated temperatures. The percent of sperm classified as alive, using fluorescein diacetate labeling, was decreased by pre-incubation and fertilization at 40.0°C. Although no difference was observed in sperm penetration rate, polyspermy tended to be increased by heat stress during fertilization. The zona pellucidae of zygotes formed following fertilization at 40.0°C for 6 hr were more sensitive to digestion with pronase. Furthermore, these zygotes exhibited higher hydrogen peroxide levels, measured by 2,7-dihydrodichlorofluorescein diacetate staining, and showed increased transcript abundance for HSPA1A, a gene involved in the heat-shock response, but decreased transcript abundance for UCHL1, a gene involved in preventing polyspermy. Results indicate that heat stress during fertilization is lethal to sperm, and causes oxidative stress, altered transcript abundance, and a defective block to polyspermy in the zygote. Thus, an increase in polyspermy is likely one cause of the reduced competency of zygotes fertilized under elevated temperatures to develop to the blastocyst stage.

  1. Post-thaw motility of frozen boar sperm does not predict success with in vitro fertilization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using cryopreserved boar sperm rather than liquid semen for in vitro fertilization (IVF) allows improved IVF consistency. However, cryopreservation of boar sperm results in reduced post-thaw motility, fertilization and embryo development. Boars are often screened on an individual basis prior to use ...

  2. Sperm DNA damage or progressive motility: which one is the better predictor of fertilization in vitro?

    PubMed

    Simon, Luke; Lewis, Sheena E M

    2011-06-01

    Sperm progressive motility has been reported to be one of the key factors influencing in vitro fertilization rates. However, recent studies have shown that sperm DNA fragmentation is a more robust predictor of assisted reproductive outcomes including reduced fertilization rates, embryo quality, and pregnancy rates. This study aimed to compare the usefulness of sperm progressive motility and DNA damage as predictive tools of in vitro fertilization rates. Here, 136 couples provided 1,767 eggs with an overall fertilization rate of 64.2%. The fertilization rate in vitro correlated with both sperm progressive motility (r² = 0.236; P = 0.002) and DNA fragmentation (r² = -0.318; P < 0.001). The relative risk of a poor fertilization rate was 9.5 times higher in sperm of men with high DNA fragmentation (>40%) compared with 2.6 times in sperm with poor motility (<40%). Further, sperm DNA fragmentation gave a higher specificity (93.3%) in predicting the fertilization rate than progressive motility (77.8%). Finally, the odds ratio to determine fertilization rate (>70%) was 4.81 (1.89-12.65) using progressive motility compared with 24.18 (5.21-154.51) using DNA fragmentation. This study shows that fertilization rates are directly dependent upon both sperm progressive motility and DNA fragmentation, but sperm DNA fragmentation is a much stronger test.

  3. Effects of prolactin on gametes and zygotes during in vitro fertilization in mice.

    PubMed

    Fukuda, A; Noda, Y; Mori, T; Mori, C; Hashimoto, H; Hoshino, K

    1988-02-01

    Hyperprolactinemia is one of the major causative factors of infertility. However, the effect of prolactin on gametes during in vitro fertilization has not been elucidated. In the present study, the effects of mouse prolactin on the motility of spermatozoa, in vitro fertilization, and in vitro development of the zygote were investigated in mice using media containing three different concentrations (10, 50, and 100 ng/ml) of mouse prolactin. The development of unfertilized and fertilized oocytes (zygotes) was not affected in vitro by prolactin regardless of the amount of prolactin added to culture media. However, the fertilizing capacity of the spermatozoa was suppressed when they were preincubated for 90 min in culture media containing prolactin at concentrations of 50 and 100 ng/ml. The motility of spermatozoa was not affected by prolactin regardless of the concentration of prolactin used for preincubation. The present study indicates that prolactin may have some effects on the capacitation process of spermatozoa in vitro. This result should be taken into consideration in in vitro fertilization and embryo transfer procedures in humans.

  4. In vitro fertilization and development of bovine oocytes matured in serum-free medium.

    PubMed

    Saeki, K; Hoshi, M; Leibfried-Rutledge, M L; First, N L

    1991-02-01

    This study was undertaken to investigate the effects of supplementation of serum (fetal calf serum), gonadotropins (LH, FSH, prolactin) and estradiol-17 beta (E2) to culture medium during in vitro maturation of bovine cumulus oocyte complexes on subsequent fertilization and development to the blastocyst stage in vitro. Serum supplementation during bovine oocyte maturation was not required but hormonal supplementation, gonadotropins (LH + FSH) and E2, enhanced the fertilizability and developmental ability of bovine oocytes matured in vitro. The addition of prolactin to maturation medium containing LH, FSH, and E2 did not further enhance frequencies of fertilization and development. PMID:2009327

  5. In vitro fertilization and development of bovine oocytes matured in serum-free medium.

    PubMed

    Saeki, K; Hoshi, M; Leibfried-Rutledge, M L; First, N L

    1991-02-01

    This study was undertaken to investigate the effects of supplementation of serum (fetal calf serum), gonadotropins (LH, FSH, prolactin) and estradiol-17 beta (E2) to culture medium during in vitro maturation of bovine cumulus oocyte complexes on subsequent fertilization and development to the blastocyst stage in vitro. Serum supplementation during bovine oocyte maturation was not required but hormonal supplementation, gonadotropins (LH + FSH) and E2, enhanced the fertilizability and developmental ability of bovine oocytes matured in vitro. The addition of prolactin to maturation medium containing LH, FSH, and E2 did not further enhance frequencies of fertilization and development.

  6. The application of the zona-free hamster egg test for the prognosis of human in vitro fertilization.

    PubMed

    Foreman, R; Cohen, J; Fehilly, C B; Fishel, S B; Edwards, R G

    1984-09-01

    The zona-free hamster egg test was carried out using spermatozoa from 15 men which consistently failed to fertilize their wives' oocytes in vitro. Spermatozoa from nine of these men fertilized hamster eggs in vitro, indicating that positive results in this assay are an unreliable guide to human in vitro fertilization. Donor spermatozoa were needed to fertilize the wife's oocytes in three of these cases. Nevertheless, the proportion of hamster egg penetration was significantly lower compared with spermatozoa from 15 men who could fertilize their wives' oocytes in vitro. The hamster assay also failed to indicate the establishment of pregnancy.

  7. Acupuncture and In Vitro Fertilization: A Retrospective Chart Review

    PubMed Central

    Opsahl, Michael S.; Taylor-Swanson, Lisa; Ackerman, Deborah L.

    2013-01-01

    Abstract Objectives In 2007, Craig et al. reported the results of a randomized controlled trial in which a standardized acupuncture protocol performed on the day of embryo transfer (ET) resulted in lower pregnancy rates after in vitro fertilization (IVF). Between 2005 and 2007, the Craig protocol was used by one of the authors (LHR) at an infertility clinic unaffiliated with the Craig et al. trial. The objective was to retrospectively review clinic records to evaluate the effect of the Craig protocol in both donor and nondonor IVF cycles on four outcomes: (1) live births; (2) biochemical pregnancies; (3) adverse outcomes; and (4) live births in nondonor cycles across age groups established by the Society for Assisted Reproductive Technology. Design The study design was a retrospective chart review. Setting The study was conducted at a private infertility clinic. Patient(s) Patients underwent fresh, donor (N=70) or nondonor (N=402) IVF-ET. Intervention(s) The Craig protocol included the following points before ET: GV-20, CV-6, ST-29, SP-8, PC-6, LV-3; Shenmen and Brain on the left ear; and Uterus and Endocrine on the right ear. After transfer the points were LI-4, SP-10, ST-36, SP-6, KI-3; Uterus and Endocrine on the left ear; and Shenmen and Brain on the right ear. Main outcome measure(s) Live births (LB) beyond 24 weeks' gestation was the main outcome measure. Result(s) In nondonor IVF cycles, there were no differences in LB across age groups (odds ratio [OR]=1.04, 95% confidence interval [CI] 0.68–1.57), biochemical pregnancies (OR=0.60, 95% CI 0.27–1.33), or adverse outcomes (OR=0.63, 95% CI 0.31–1.26). In donor cycles, LB were higher in the acupuncture group (relative risk=1.31, 95% CI 1.02–1.71). Conclusions In this observational study, the Craig protocol was not found to lower IVF LB. In fact, the Craig protocol was associated with higher LB in donor cycles. These findings should be considered cautiously because more adequately powered, randomized

  8. Effects of exposure to zearalenone on porcine oocytes and sperm during maturation and fertilization in vitro.

    PubMed

    Sambuu, Rentsenkhand; Takagi, Mitsuhiro; Namula, Zhao; Otoi, Takeshige; Shiga, Satoshi; Rodrigues Dos Santos, Regiane; Fink-Gremmels, Johanna

    2011-09-01

    The influence of acute exposure to zearalenone (ZEN) on porcine oocyte maturation, fertilization or sperm penetration ability during both in vitro maturation and fertilization was evaluated. First, oocytes were cultured in ZEN-containing (0-1000 µg/l) maturation medium and then fertilized. The oocytes maturing in vitro without ZEN were then fertilized in ZEN-containing fertilization medium. The maturation rates of oocytes and penetration ability of sperm decreased significantly in the presence of 1000 µg/l of ZEN. However, neither increases in the rates of degeneration and DNA fragmentation of oocytes nor reductions in normal and polyspermic fertilization were observed. ZEN did not affect the sperm penetration rates; however, 1000 µg/l ZEN had positive effects on normal and polyspermic fertilization rates. Therefore, it can be suggested that an acute exposure of porcine oocytes during maturation and of oocytes and sperm during fertilization to ZEN up to 1000 µg/l may not affect the fertility of the oocytes.

  9. Treatment independent pregnancy with operative laparoscopy for endometriosis in an in vitro fertilization program.

    PubMed

    Damewood, M D; Rock, J A

    1988-09-01

    Thirty-nine patients with Stage I to IV endometriosis and at least 5 years of primary infertility were enrolled in the Johns Hopkins In Vitro Fertilization Program. At the time of laparoscopic oocyte retrieval, operative endoscopy with lysis of adhesions, fulguration or resection of pelvic endometriosis, or enucleation of ovarian endometriomas was performed. Although the in vitro fertilization-embryo transfer cycle did not result in pregnancy, 12 patients (28%) conceived within 10 months of the operative laparoscopic procedure. Nine of the pregnancies occurred in patients with Stage I to II endometriosis.

  10. In vitro fertilization of water buffalo follicular oocytes and their ability to cleave in vitro.

    PubMed

    Suzuki, T; Singla, S K; Sujata, J; Madan, M L

    1992-12-01

    Water buffalo (Murrah) oocytes were collected from ovaries obtained from a local slaughterhouse. They were classified according to the character of the cumulus cells under a stereomicroscope and then cultured in 25 mM Hepes buffered tissue culture medium-199 (TCM-199) supplemented with 5% estrous water buffalo serum in an atmosphere containing 5% CO2 in air at 39 degrees C. After 20 to 24 hours of in vitro maturation, the oocytes were cultured at 38.5 degrees C in TCM-199 supplemented with 1% estrous water buffalo serum and in an atmosphere containing 5% CO2 in air. Oocytes with compact and dense cumulus cells cleaved significantly further (P<0.01, 67.3%, 33/49) than those with fair, partially denuded oocytes with thin cumulus layers (27.5%, 25/91) or small remnants of cumulus cells and poor naked oocytes (3/100). A substantial variation in fertilization and developmental rates (16.0 to 43.8%) was observed among 4 different bulls. Late morulae were transferred nonsurgically into 14 buffalo recipients on Day 6 or 7 of their estrous cycle. One recipient was diagnosed to be pregnant by palpation per rectum on Day 60 and delivered a calf in October 1991.

  11. The hemizona assay (HZA): a predictor of human sperm fertilizing potential in in vitro fertilization (IVF) treatment.

    PubMed

    Franken, D R; Oehninger, S; Burkman, L J; Coddington, C C; Kruger, T F; Rosenwaks, Z; Acosta, A A; Hodgen, G D

    1989-02-01

    The hemizona assay (HZA) was developed to assess human sperm fertilizing potential. This blinded study investigated the relationship between sperm binding to the hemizona and in vitro fertilization (IVF) success (36 patients). Nonliving human oocytes were recovered from excised ovaries and stored. Each zona pellucida was cut into equal hemispheres by micromanipulation. For the HZA, one droplet exposed a hemizona to abnormal spermatozoa, while the control droplet contained the matching hemizona and spermatozoa from normal semen. After 4 hr, the number of tightly bound spermatozoa was counted. Binding to the hemizona was significantly higher for those having IVF success (mean of 36.1 +/- 7, versus 10.4 +/- 4 from the failure group; P less than 0.05). Fewer sperm from the failure group had a strictly normal morphology (3.2 versus 12.7%; P less than 0.05, Kruger method). Tight zona binding was significantly correlated with the percentage motile sperm, percentage normal morphology, and seminal sperm concentration. These results enhanced our confidence that the HZA is diagnostic for identification of patients at high risk of failing to achieve fertilization in vitro.

  12. Transient exposure to calcium ionophore enables in vitro fertilization in sterile mouse models

    PubMed Central

    Navarrete, Felipe A.; Alvau, Antonio; Lee, Hoi Chang; Levin, Lonny R.; Buck, Jochen; Leon, Patricia Martin-De; Santi, Celia M.; Krapf, Dario; Mager, Jesse; Fissore, Rafael A.; Salicioni, Ana M.; Darszon, Alberto; Visconti, Pablo E.

    2016-01-01

    Mammalian sperm acquire fertilizing capacity in the female tract in a process called capacitation. At the molecular level, capacitation requires protein kinase A activation, changes in membrane potential and an increase in intracellular calcium. Inhibition of these pathways results in loss of fertilizing ability in vivo and in vitro. We demonstrated that transient incubation of mouse sperm with Ca2+ ionophore accelerated capacitation and rescued fertilizing capacity in sperm with inactivated PKA function. We now show that a pulse of Ca2+ ionophore induces fertilizing capacity in sperm from infertile CatSper1 (Ca2+ channel), Adcy10 (soluble adenylyl cyclase) and Slo3 (K+ channel) KO mice. In contrast, sperm from infertile mice lacking the Ca2+ efflux pump PMACA4 were not rescued. These results indicate that a transient increase in intracellular Ca2+ can overcome genetic infertility in mice and suggest this approach may prove adaptable to rescue sperm function in certain cases of human male infertility. PMID:27627854

  13. Transient exposure to calcium ionophore enables in vitro fertilization in sterile mouse models.

    PubMed

    Navarrete, Felipe A; Alvau, Antonio; Lee, Hoi Chang; Levin, Lonny R; Buck, Jochen; Leon, Patricia Martin-De; Santi, Celia M; Krapf, Dario; Mager, Jesse; Fissore, Rafael A; Salicioni, Ana M; Darszon, Alberto; Visconti, Pablo E

    2016-01-01

    Mammalian sperm acquire fertilizing capacity in the female tract in a process called capacitation. At the molecular level, capacitation requires protein kinase A activation, changes in membrane potential and an increase in intracellular calcium. Inhibition of these pathways results in loss of fertilizing ability in vivo and in vitro. We demonstrated that transient incubation of mouse sperm with Ca(2+) ionophore accelerated capacitation and rescued fertilizing capacity in sperm with inactivated PKA function. We now show that a pulse of Ca(2+) ionophore induces fertilizing capacity in sperm from infertile CatSper1 (Ca(2+) channel), Adcy10 (soluble adenylyl cyclase) and Slo3 (K(+) channel) KO mice. In contrast, sperm from infertile mice lacking the Ca(2+) efflux pump PMACA4 were not rescued. These results indicate that a transient increase in intracellular Ca(2+) can overcome genetic infertility in mice and suggest this approach may prove adaptable to rescue sperm function in certain cases of human male infertility. PMID:27627854

  14. The influence of transient hyperprolactinemia on hormonal parameters, oocyte recovery, and fertilization rates in in vitro fertilization.

    PubMed

    Gonen, Y; Casper, R F

    1989-06-01

    A detrimental effect of transient elevation of plasma prolactin (PRL) during in vitro fertilization (IVF) has not been proven; however, treatment with a dopamine agonist has been suggested. The present study was undertaken to determine if transient, midcycle hyperprolactinemia exerted a deleterious effect on the number of oocytes retrieved or on fertilization of oocytes in vitro. Fifty-three infertile patients with midcycle hyperprolactinemia (PRL greater than 20 micrograms/liter) during ovarian hyperstimulation for IVF were compared with 53 matched controls who remained normoprolactinemic. Mean (+/- SE) serum PRL levels on the day after hCG were significantly higher in the study group (29.5 +/- 1 micrograms/liter) than in the control (13.1 +/- 0.5 microgram/liter) (P less than 0.0005), whereas the mean estradiol (E2) concentrations on the same day were not significantly different (4822 +/- 287 and 4492 +/- 269 pmol/liters, respectively). Fertilization rates (72 +/- 4 and 70 +/- 4%, respectively) and the mean number of oocytes recovered (4.2 +/- 0.3 and 3.7 +/- 0.3, respectively) did not differ between the two groups. No correlation was observed between serum PRL and E2 levels, fertilization rates, or the number of oocytes retrieved in either group. Eleven patients with elevated PRL levels as a result of ovarian hyperstimulation were treated with 2.5 mg bromocriptine daily during the next IVF cycle. Serum PRL levels were significantly lower in the treated (5.6 +/- 1.8 micrograms/liter) than in the untreated cycles (35.6 +/- 3.1 micrograms/liter) (P less than 0.0005), whereas serum E2 concentrations did not differ.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. In vitro fertilization in the Netherlands: experiences and opinions of Dutch women.

    PubMed

    Holmes, H B; Tymstra, T

    1987-04-01

    We sent written questionnaires to 93 Dutch women [73 current and potential clients of the Rotterdam in vitro fertilization (IVF) program and 20 fertile nonusers] to determine the motivation for participation in IVF, patients' reactions to treatment, and views of concerned laypersons on current ethical/social issues. Seventy-eight (84%) completed the questionnaires. A large majority was strongly positive toward the having of children and toward IVF, but fewer fertile than infertile women (P less than 0.05) believed that "a child of one's own" was a right and that government insurance ought to cover IVF. Nearly all the patients (with or without successful pregnancies) were very satisfied with their clinical treatment and would recommend IVF. A large majority of all the women favored gamete donation and surrogate gestation, but fewer fertile than infertile women approved experiments with "spare" embryos or implanting donor embryos.

  16. Human sperm devoid of germinal angiotensin-converting enzyme is responsible for total fertilization failure and lower fertilization rates by conventional in vitro fertilization.

    PubMed

    Li, Le-Jun; Zhang, Feng-Bin; Liu, Shu-Yuan; Tian, Yong-Hong; Le, Fang; Wang, Li-Ya; Lou, Hang-Ying; Xu, Xiang-Rong; Huang, He-Feng; Jin, Fan

    2014-06-01

    In conventional in vitro fertilization (IVF), complete failure of fertilization occurs in 5% to 15% of treatments. Although the causes may be unclear, sperm defects appear to be the major contributor. However, a convincing test is not yet available that can predict the risk of fertilization failure. In this study, we found that germinal angiotensin-converting enzyme (gACE) (also called testicular ACE) was undetectable in sperm from patients who had total fertilization failure (TFF) and lower fertilization rates (LFRs) by IVF based on Western blot and indirect immunofluorescence analyses. Additionally, almost all of the patients without gACE on sperm (23 of 25) manifested a TT genotype of the rs4316 single-nucleotide polymorphism of ACE. Overall, our results indicate that the absence of gACE expression is responsible for TFF and LFRs by IVF. The rs4316 polymorphism of ACE might be associated with infertility in those patients. We conclude that sperm lacking gACE may be recognized before commencing IVF and that the patients may be directed instead to consider intracytoplasmic sperm injection.

  17. In vitro fertilization in Japan — Early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology —

    PubMed Central

    SUZUKI, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world’s first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe.1) In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine.2,3) IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient’s uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART. PMID:24814992

  18. In vitro fertilization in Japan - early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology.

    PubMed

    Suzuki, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world's first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe. In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine. IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient's uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART. PMID:24814992

  19. Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro

    PubMed Central

    2013-01-01

    Background Good quality spermatozoa are important to achieve fertilization, viable embryos and offspring. Single Layer Centrifugation (SLC) through a colloid (Androcoll-P) selects good quality spermatozoa. However, it has not been established previously whether porcine spermatozoa selected by this method maintain their fertility. Methods The semen was prepared either by SLC or by standard centrifugation (control) and used for in vitro fertilization (IVF) at oocyte:spermatozoa ratios of 1:50; 1:100 and 1:300 (or 4 x 103, 8 x 103 and 24 x 103 spermatozoa/ml) to evaluate their subsequent ability to generate blastocysts. In addition, sperm motility was assessed by computer assisted sperm motility analysis. Results Total and progressive motility were significantly higher in sperm samples prepared by SLC compared to uncentrifuged samples. Sperm binding ability, polyspermy, cleavage and blastocyst rates were affected by the oocyte:sperm ratio, but not by sperm treatment. Conclusion The use of SLC does not adversely affect the in vitro fertilizing and embryo-generating ability of the selected spermatozoa compared to their unselected counterparts, but further modifications in the IVF conditions would be needed to improve the monospermy in IVF systems. Since SLC did not appear to have a negative effect on sperm fertilizing ability, and may in fact select for spermatozoa with a greater potential for fertilization, an in vivo trial to determine the usefulness of this sperm preparation technique prior to artificial insemination is warranted. PMID:23497680

  20. Drug delivery for in vitro fertilization: rationale, current strategies and challenges.

    PubMed

    Janát-Amsbury, Margit M; Gupta, Kavita M; Kablitz, Caroline D; Peterson, C Matthew

    2009-08-10

    In vitro fertilization has experienced phenomenal progress in the last thirty years and awaits the additional refinement and enhancement of medication delivery systems. Opportunity exists for the novel delivery of gonadotropins, progesterone and other adjuvants. This review highlights the rationale for various medications, present delivery methods and introduces the status of novel ideas and possibilities.

  1. Adoption Actions and Attitudes of Couples Seeking In Vitro Fertilization: An Exploratory Study.

    ERIC Educational Resources Information Center

    Williams, Linda S.

    1992-01-01

    Examined adoption actions and attitudes of 16 childless women, and husbands of 14 of the women, who applied for or underwent in vitro fertilization (IVF). Found that IVF and adoption were sought concurrently by most and that wives were more in favor of adoption than were husbands. (Author/NB)

  2. Novel and traditional traits of frozen-thawed porcine sperm related to in vitro fertilization success

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryopreserved semen allows the use of single ejaculates for repeated analyses, potentially improving in vitro fertilization (IVF) consistency by eliminating inter-ejaculate variability observed with fresh semen. However, the freezing and thawing processes result in compromised sperm function and IVF...

  3. Rescue in vitro fertilization method for legacy stock of frozen mouse sperm.

    PubMed

    Nakagata, Naomi; Takeo, Toru; Fukumoto, Kiyoko; Haruguchi, Yukie; Kondo, Tomoko; Takeshita, Yumi; Nakamuta, Yuko; Umeno, Tomoko; Tsuchiyama, Shuuji

    2014-04-24

    Sperm cryopreservation has been widely adopted for maintenance of the genetically engineered mouse (GEM). The cryopreserved sperm are being exchanged among many institutes worldwide. However, the recipients are not always able to obtain high fertilization rates with the frozen sperm shipped from senders. In this study, we cryopreserved mouse sperm via various methods and performed in vitro fertilization (IVF) in which the combination of methyl-beta-cyclodextrin for sperm preincubation and reduced glutathione for insemination was used (the MBCD-GSH IVF). In addition, frozen sperm sent from the Jackson Laboratory (USA) were thawed and used for IVF in the same manner. The fertilization rates of both the sperm cryopreserved via the methods applied in some countries and the cryopreserved GEM sperm improved when used with the MBCD-GSH IVF method. Therefore, we strongly believe that the MBCD-GSH IVF method brings about relatively high fertilization rates with any strain of frozen mouse sperm.

  4. Improvement in in vitro fertilization outcome following in vivo synchronization of oocyte maturation in mice.

    PubMed

    Taiyeb, Ahmed M; Muhsen-Alanssari, Saeeda A; Dees, W L; Ridha-Albarzanchi, Mundhir T; Kraemer, Duane C

    2015-04-01

    Synchronization of oocyte maturation in vitro has been shown to produce higher in vitro fertilization (IVF) rates than those observed in oocytes matured in vitro without synchronization. However, the increased IVF rates never exceeded those observed in oocytes matured in vivo without synchronization. This study was therefore designed to define the effect of in vivo synchronization of oocyte maturation on IVF rates. Mice were superovulated and orally treated with 7.5 mg cilostazol (CLZ), a phosphodiesterase 3A (PDE3A) inhibitor, to induce ovulation of immature oocytes at different stages depending on frequency and time of administration of CLZ. Mice treated with CLZ ovulated germinal vesicle (GV) or metaphase I (MI) oocytes that underwent maturation in vitro or in vivo (i.e. in the oviduct) followed by IVF. Superovulated control mice ovulated mature oocytes that underwent IVF directly upon collection. Ovulated MI oocytes matured in vitro or in vivo had similar maturation rates but significantly higher IVF rates, 2-4 cell embryos, than those observed in control oocytes. Ovulated GV oocytes matured in vitro showed similar maturation rates but significantly higher IVF rates than those observed in control oocytes. However, ovulated GV oocytes matured in vivo had significantly lower IVF rates than those noted in control oocytes. It is concluded that CLZ is able to synchronize oocyte maturation and improve IVF rates in superovulated mice. CLZ may be capable of showing similar effects in humans, especially since temporal arrest of human oocyte maturation with other PDE3A inhibitors in vitro was found to improve oocyte competence level. The capability of a clinically approved PDE3A inhibitor to improve oocyte fertilization rates in mice at doses extrapolated from human therapeutic doses suggests the potential scenario of the inclusion of CLZ in superovulation programs. This may improve IVF outcomes in infertile patients.

  5. Disentangling causal paths between obesity and in vitro fertilization outcomes: an intractable problem?

    PubMed

    Howards, Penelope P; Cooney, Maureen A

    2008-06-01

    Research questions should be appropriate for the available data. Disentangling causal paths between obesity and in vitro fertilization outcomes may be an intractable problem, but other research questions of clinical importance can be addressed through appropriate study designs and analytic methods.

  6. In Vitro Fertilization and the Family: Quality of Parenting, Family Functioning, and Child Psychosocial Adjustment.

    ERIC Educational Resources Information Center

    Hahn, Chun-Shin; DiPietro, Janet A.

    2001-01-01

    Examined associations between in vitro fertilization (IVF) and parenting quality, family functioning, and emotional/behavioral adjustment of 3- to 7-year-olds. Found that IVF mothers reported greater protectiveness than mothers of naturally conceived children. Teachers rated IVF mothers as displaying greater warmth but not overprotective or…

  7. In vitro pollination of maize (Zea mays L.) - Proof of double fertilization.

    PubMed

    Havel, L; Novák, F J

    1981-08-01

    Isolated ovules from the maize homozygous recessive brown midrib (bm3) were in vitro pollinated by pollen carrying the dominant allele - and the purple embryo marker (PEM). The colour characters of the embryos and kernels corresponded to the results of control pollination and confirmed the process of double fertilization. The question whether the method is useful for obtaining haploids is discussed. PMID:24258750

  8. Collection of gametes from live axolotl, Ambystoma mexicanum, and standardization of in vitro fertilization.

    PubMed

    Mansour, N; Lahnsteiner, F; Patzner, R A

    2011-01-15

    This study established the first protocol for collection of gametes from live axolotl, Ambystoma mexicanum, by gentle abdominal massage and in vitro fertilization. To stimulate spermiation and ovulation, human chorionic gonadotrophin (hCG) and Ovopel pellets, which are commercially used to stimulate spawning in fish, were tested. The hCG was more effective than Ovopel pellets and yielded a higher semen volume in the injected males and a shorter response time in the females. Collected semen by this method was already motile and fertile. Fertile eggs could be collected in 3-4 successive collection times after the female has started the typical spawning behaviour. The fertilization condition that yielded the highest hatching rate was mixing semen with eggs before the addition of a fertilization saline solution (20 mmol/l NaCl, 1 mmol/l KCl, 1 mmol/l Mg(2)SO(4), 1 mmol Ca(2)Cl, 3 mmol NaHCO(3), 10 mmol/l Tris, pH 8.5 - Osmolality = 65 mosmol/kg). When the pH of the fertilization solution was increased to ≥ 10, the hatching rate was significantly increased. The use of fertilization solutions with osmolalities of ≥ 150 and ≥ 182 were accompanied with a significant decrease in hatching rates and the appearance of deformed larvae, respectively. In conclusion, a reliable protocol for gamete collection from live axolotl is established as a laboratory model of in vitro fertilization for urodele amphibians. This protocol may be transferable to endangered urodeles.

  9. Successful fertilization, embryo development, and pregnancy in human in vitro fertilization (IVF) using a chemically defined culture medium containing no protein.

    PubMed

    Caro, C M; Trounson, A

    1986-08-01

    A randomized control trial involving the fertilization and culture of human embryos in culture medium (T6) containing either 10% maternal serum or no protein or amino acid supplement was carried out to assess the effect of deletion from culture of all fixed nitrogen on fertilization, embryo development, and embryo viability. There was no difference in fertilization rates (68 vs 69%), development of apparently normal embryos (96 vs 97%), pregnancy rate (18 vs 14%), or birth rate (13 vs 11%) between protein-containing and protein-free media. Deletion of protein from the culture medium may enable the constitution of more appropriate and defined culture media for human in vitro fertilization (IVF).

  10. In vitro fertilization of mouse ova by spermatozoa exposed isothermally to radio-frequency radiation

    SciTech Connect

    Cleary, S.F.; Liu, L.M.; Graham, R.; East, J. )

    1989-01-01

    Mouse spermatozoa were exposed in vitro for 1 h to 27- or 2,450-MHz CW RF radiation at SARs of 0 to 90 W/kg under isothermal (37 +/- 0.2 degrees C) conditions. Exposure at either frequency to RF radiation at SARs of 50 W/kg or greater resulted in a statistically significant reduction in the ability of irradiated sperm to fertilize mouse ova in vitro (P less than .05). Over the range of SARs there was no apparent difference in the effects of 27- vs. 2,450-MHz RF radiation. There were no readily detectable exposure effects on spermatozoan morphology, ultrastructure, or capacitation. The reduction of in vitro fertilization is attributed to a direct effect of RF radiation on spermatozoa rather than to heating.

  11. Varicocele management in the era of in vitro fertilization/intracytoplasmic sperm injection

    PubMed Central

    Pathak, Piyush; Chandrashekar, Aravind; Hakky, Tariq S; Pastuszak, Alexander W

    2016-01-01

    Varicocele is the most common surgically treatable cause of male infertility, and often results in alterations in semen parameters, sperm DNA damage, and changes to the seminal milieu. Varicocele repair can result in improvement in these parameters in the majority of men with clinical varicocele; data supporting repair in men with subclinical varicocele are less definitive. In couples seeking fertility using assisted reproductive technologies (ARTs), varicocele repair may offer improvement in semen parameters and sperm health that can increase the likelihood of successful fertilization using techniques such as in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), or may decrease the level of ART needed to achieve successful pregnancy. Male infertility is an indicator of general male health, and evaluation of the infertile male with an eye toward future health can facilitate optimal screening and treatment of these men. Furthermore, varicocele may represent a progressive lesion, offering an argument for its repair, although this is currently unclear. PMID:27030086

  12. Mouse embryo quality control for toxicity determination in the Norfolk in vitro fertilization program.

    PubMed

    McDowell, J S; Swanson, R J; Maloney, M; Veeck, L

    1988-06-01

    Ongoing quality control is necessary as part of the maintenance and improvement of a successful human in vitro fertilization (IVF) program. Using a mouse quality-control culture system, several instrument preparation protocols were reevaluated to determine their efficiency in the control or elimination of potential toxicity. Dilute concentrations of urine and endometrial fluid were also tested. Medium rinsed through laparoscope and aspiration needles failed to support embryo development. This effect was reversed in needles that were pretreated with rinses of Dulbecco's phosphate-buffered saline. Endometrial fluid demonstrated no obvious toxic effect, but urine-exposed embryos arrested in the two-cell state. The importance of periodic evaluation of materials and their pretreatment before use in in vitro fertilization of human oocytes is essential to ensure control of potentially toxic substances.

  13. The effect of glucocorticoids on mouse oocyte in vitro maturation and subsequent fertilization and embryo development.

    PubMed

    González, Raquel; Ruiz-León, Yolanda; Gomendio, Montserrat; Roldan, Eduardo R S

    2010-02-01

    Increased glucocorticoid levels, due to medical therapy or stress-related, may affect reproduction via the hypothalamus-pituitary-axis or directly at the oocyte level. We examined the effects of natural (corticosterone) or synthetic (dexamethasone) glucocorticoids on mouse oocyte maturation and underlying changes in extracellular signal-regulated kinase (ERK) phosphorylation patterns. Fertilization and progression up to the blastocyst stage were also evaluated. Oocytes were exposed to corticosterone or dexamethasone (0, 0.25, 2.5, 25 or 250microM) for 17h during in vitro maturation. After maturation, ERK-1/2 activation in oocytes was assessed by SDS-PAGE and immunoblotting, and fertilization and developmental capacity were examined in vitro. Corticosterone exposure during oocyte maturation significantly decreased progression to metaphase II, fertilization and embryo development at the highest concentration. Corticosterone caused a concentration-dependent inhibition of ERK-1/2 activation, with the highest concentration resulting in considerable inhibition of oocyte ERK-1/2 phosphorylation and no blastocyst development. In contrast, dexamethasone had no effect on maturation, fertilization and cleavage, and no effect was seen on ERK-1/2 phosphorylation. Based on these in vitro findings, high glucocorticoid levels may have consequences for subsequent development, although a short exposure to physiologic or stress-related glucocorticoid levels may not represent a hazard to meiosis progression of the oocyte.

  14. The effect of glucocorticoids on mouse oocyte in vitro maturation and subsequent fertilization and embryo development.

    PubMed

    González, Raquel; Ruiz-León, Yolanda; Gomendio, Montserrat; Roldan, Eduardo R S

    2010-02-01

    Increased glucocorticoid levels, due to medical therapy or stress-related, may affect reproduction via the hypothalamus-pituitary-axis or directly at the oocyte level. We examined the effects of natural (corticosterone) or synthetic (dexamethasone) glucocorticoids on mouse oocyte maturation and underlying changes in extracellular signal-regulated kinase (ERK) phosphorylation patterns. Fertilization and progression up to the blastocyst stage were also evaluated. Oocytes were exposed to corticosterone or dexamethasone (0, 0.25, 2.5, 25 or 250microM) for 17h during in vitro maturation. After maturation, ERK-1/2 activation in oocytes was assessed by SDS-PAGE and immunoblotting, and fertilization and developmental capacity were examined in vitro. Corticosterone exposure during oocyte maturation significantly decreased progression to metaphase II, fertilization and embryo development at the highest concentration. Corticosterone caused a concentration-dependent inhibition of ERK-1/2 activation, with the highest concentration resulting in considerable inhibition of oocyte ERK-1/2 phosphorylation and no blastocyst development. In contrast, dexamethasone had no effect on maturation, fertilization and cleavage, and no effect was seen on ERK-1/2 phosphorylation. Based on these in vitro findings, high glucocorticoid levels may have consequences for subsequent development, although a short exposure to physiologic or stress-related glucocorticoid levels may not represent a hazard to meiosis progression of the oocyte. PMID:19733225

  15. [Is an act of human love the in vitro fertilization? A proposal ethical analysis].

    PubMed

    García Sánchez, Emilio

    2014-01-01

    Since 1978, when the first test tube baby, Louis Brown, was born, thousands of children have been born every year through in vitro fertilization. Many families keep attending fertility clinics in order to receive some treatment for their infertility problems and have a child. Children born in this way are worthy human beings. Their parents love them and devote themselves to their children admirably, showing real parental love. However, does this loving kindness justify, from an ethical point of view, any way of desiring and having a son or daughter? Is it really an act of human love to long for a child and satisfy this desire using artificial methods? Is it equally human and worthy to wish them choosing in vitro fertilization than to wish them through an intimate and loving relationship, in which the child emerges as a result of interpersonal donation? I answer these questions by analyzing the ethics proposal formulated by Rhonheimer and Carrasco de Paula. In short, only the intimate and loving sexual union between a man and a woman -as long as it is unconditional love- may be the dignity cause of the existence of a human being. And such union and unconditional requirement are absent in vitro fertilization.

  16. Postpartal dissection of all coronary arteries in an in vitro-fertilized postmenopausal woman.

    PubMed

    Karadag, Bilgehan; Roffi, Marco

    2009-01-01

    Myocardial infarction complicates approximately 1 in 10,000 pregnancies. Although coronary artery dissection is the leading cause of pregnancy-related myocardial infarction during the postpartum period, the pathogenesis of coronary dissection during this period remains uncertain.Herein, we report the case of a 52-year-old black postmenopausal woman with no apparent cardiovascular risk factors who gave birth to twins after in vitro fertilization. Ten days after delivery, she presented with an acute coronary syndrome. Coronary angiography revealed dissection of all 3 coronary arteries. Despite aggressive medical management, the patient experienced recurrent myocardial ischemia. Repeat coronary angiography revealed progression of the dissection process, which required urgent coronary artery bypass surgery. The patient's postoperative course was uneventful. To our knowledge, this report is the 1st description of pregnancy-associated coronary artery dissections in a postmenopausal woman, and the 1st such event in a pregnancy that resulted from in vitro fertilization.

  17. Transient hyperprolactinemia has no effect on endocrine response and outcome in in vitro fertilization (IVF).

    PubMed

    Pattinson, H A; Taylor, P J; Fleetham, J A; Servis, S A

    1990-04-01

    Transient rises in plasma prolactin levels can be observed during the late follicular phases of both natural and stimulated cycles. It has been suggested that such a phenomenon might adversely affect the success of in vitro fertilization. This prospective study was designed to assess the effect of transient rises in prolactin levels on the endocrine response to ovarian stimulation and the outcome of in vitro fertilization treatment. A total of 90 treatment cycles in 87 couples was studied. Prolactin was measured in the mid and late follicular phases of the cycles. During the study period, 24 pregnancies occurred. There were no differences in those cycles in which pregnancy did or did not occur in either mid or late follicular prolactin levels. Neither the initial level nor the percentage rise in prolactin during the stimulation had any effect on the peak estradiol level achieved, the numbers of follicles seen, the number of eggs retrieved, or the incidence or outcome of pregnancy. It was concluded that transient hyperprolactinemia is of no significance in ovarian stimulation for in vitro fertilization.

  18. Buffalo and cattle hybrid embryo development is decreased by caffeine treatment during in vitro fertilization.

    PubMed

    Tatham, B G; Feehan, T; Pashen, R

    2003-02-01

    Water buffalo are renowned for difficulties in the implementation of assisted reproductive technologies, with both males and females being problematic. In this study, we used cattle oocytes to assess the effect of treatments with heparin and caffeine on buffalo spermatozoa and subsequent fertilization and embryo development in vitro. There was no significant difference between buffalo and bovine spermatozoa in the events associated with fertilization. Fertilization of cattle oocytes with buffalo spermatozoa resulted in 7.8% of oocytes developing into hybrid embryos. A difference in the developmental capability of hybrid embryos compared with the cattle control was observed. This has not been previously reported. The subsequent transfer of a limited number of hybrid embryos did not produce a viable pregnancy. However, control treatments in this experiment also failed to achieve pregnancy, so objective data is not available to provide conclusions about the developmental competence of the buffalo and cattle hybrid embryos. Optimal spermatozoa capacitation treatments achieved 61% fertilization and 21% zygote cleavage into two cell embryos. There was no significant difference in fertilization or development due to heparin or spermatozoa concentrations. However, treatment of buffalo and cattle spermatozoa with caffeine significantly decreased embryo cleavage but also tended to decrease embryo development to the blastocyst stage. These studies suggest that problems with reproduction in buffalo may reside with biological mechanisms associated with the oocyte that are often complicated by poor male reproductive performance. Selection for bull fertility would prevent some of these complications.

  19. Ureteral lesion secondary to vaginal ultrasound follicular puncture for oocyte recovery in in-vitro fertilization.

    PubMed

    Coroleu, B; Lopez Mourelle, F; Hereter, L; Veiga, A; Calderón, G; Martinez, F; Carreras, O; Barri, P N

    1997-05-01

    Techniques of oocyte retrieval have progressed from laparoscopy to transvaginal follicular aspiration under ultrasonographic control. This highly efficient method, routinely used nowadays, is not free of complications. We present a case of a ureteral lesion secondary to vaginal ultrasound follicular puncture for oocyte recovery in in-vitro fertilization. Despite the surgical procedure to reimplant the ureter, the patient achieved a twin pregnancy which is ongoing uneventfully.

  20. IVF versus ICSI for the fertilization of in-vitro matured human oocytes.

    PubMed

    Walls, M; Junk, S; Ryan, J P; Hart, R

    2012-12-01

    Traditional dogma suggests that intracytoplasmic sperm injection (ICSI) should be performed to ensure successful oocyte fertilization in an in-vitro maturation (IVM) cycle. This study postulated that there would be no difference in the fertilization rate when ICSI was compared with IVF. This hypothesis was tested in a randomized trial of IVF versus ICSI in IVM. A total of 150 immature oocytes were collected in eight cycles of IVM for patients diagnosed with polycystic ovarian syndrome (PCOS). Patients were primed with minimal FSH before transvaginal oocyte aspiration. Sibling oocytes were inseminated by 50% IVF and 50% ICSI. There was no significant difference in fertilization, useable or total blastocyst development between the two insemination technique groups. Clinical pregnancy results for combined fresh and cryopreserved transfers were identical between the two insemination techniques with a total of two fresh and five cryopreserved IVF-inseminated embryos resulting in three clinical pregnancies (42.9%) and five fresh and two cryopreserved ICSI-derived embryos resulting in three clinical pregnancies (42.9%). This research has shown IVF to be a legitimate fertilization technique for IVM oocytes in PCOS patients and provides a greater awareness of the use of a fertilization method previously not utilized with IVM. In-vitro maturation (IVM) is an alternative treatment method to traditional IVF. Due to the minimal use of stimulating hormones in this treatment, IVM has a lower risk of ovarian hyperstimulation syndrome, it can be used for fertility preservation in cancer patients and it is more cost conservative. Early research into the effects of IVM showed a hardening effect on the membrane surrounding the egg (the zona pellucida). It was initially believed that, to overcome this hardening in order to allow the egg to be fertilized, spermatozoa would need to be injected into the egg using intracytoplasmic sperm injection. Due to recent advances in hormonal

  1. In vitro fertilization experiments using sockeye salmon reveal that bigger eggs are more fertilizable under sperm limitation

    PubMed Central

    Macfarlane, Christopher P.; Hoysak, Drew J.; Liley, N. Robin; Gage, Matthew J.G.

    2009-01-01

    Although theory and widespread evidence show that the evolution of egg size is driven primarily by offspring and maternal fitness demands, an additional explanation invokes sperm limitation as a selective force that could also influence egg size optima. Levitan proposed that constraints from gamete encounter in external fertilization environments could select for enlargement of ova to increase the physical size of the fertilization target. We test this theory using in vitro fertilization experiments in an externally fertilizing fish. Sockeye salmon (Onchorhyncus nerka) females show considerable between-individual variation in ovum size, and we explored the consequences of this natural variation for the fertilization success of individual eggs under conditions of sperm limitation. By engineering consistent conditions where in vitro fertilization rate was always intermediate, we were able to compare the sizes of fertilized and unfertilized eggs across 20 fertilization replicates. After controlling for any changes in volume through incubation, results showed that successfully fertilized eggs were significantly larger than the eggs that failed to achieve fertilization. Under conditions without sperm limitation, fertility was unaffected by egg size. Our findings therefore support Levitan's theory, demonstrating empirically that some element of egg size variation could be selected by fertilization demands under sperm limitation. However, further research on sperm limitation in natural spawnings is required to assess the selective importance of these results. PMID:19364734

  2. Male reproductive traits, semen cryopreservation, and heterologous in vitro fertilization in the bobcat (Lynx rufus).

    PubMed

    Gañán, N; González, R; Sestelo, A; Garde, J J; Sánchez, I; Aguilar, J M; Gomendio, M; Roldan, E R S

    2009-08-01

    There is limited information on bobcat ejaculate traits and sperm cryopreservation and fertilizing ability. Bobcats were electroejaculated under general anesthesia in November (autumn) and April (spring), and endocrine and sperm traits were characterized. Testosterone (mean+/-SEM: 0.90+/-0.15 ng/mL) was not different between sampling times, but cortisol (average: 13.95+/-1.73 microg/dL) was significantly higher in April. Average number of spermatozoa was 10.0+/-3.4 x 10(6) sperm/ejaculate, with values being significantly higher in April. Sperm motility (average 55.7+/-5.8% motile sperm) was not different between sampling times. The proportion of normal spermatozoa in the ejaculate (average: 14.7+/-2.1%) was significantly higher in April, but the percentage of spermatozoa with intact acrosomes (average: 43.7+/-3.8%) was significantly higher in autumn. Spermatozoa were cryopreserved in a Tes-Tris-based diluent (TEST) or Biladyl, both containing 20% egg yolk and 4% glycerol. Diluted sperm were loaded into straws, refrigerated using a programmable thermoblock with a dry chamber, frozen in nitrogen vapors, thawed, and incubated in F-10 medium with 5% fetal bovine serum for up to 3h. After cryopreservation in TEST, there were about 50% motile sperm upon thawing, and survival was high during incubation post-thaw. Cryopreservation in Biladyl led to similar results, but motility decreased substantially during incubation post-thaw. Bobcat spermatozoa fertilized domestic cat oocytes matured in vitro. Fertilization rates were higher for sperm collected in April and cryopreserved in TEST (46%) than for those cryopreserved using Biladyl (<3%). Fertilized oocytes cleaved in culture, and some (27%) reached the morula stage. This study has allowed us to gain further baseline information on bobcat reproduction, explore sperm cryopreservation conditions, and show that fertilizing capacity can be tested using in vitro-matured cat oocytes. These results will be important for future

  3. Effects of lead on the male mouse as investigated by in vitro fertilization and blastocyst culture

    SciTech Connect

    Johansson, L.; Sjoeblom, P.; Wide, M.

    1987-02-01

    Long-term exposure of male mice to inorganic lead (lead chloride, 1 g/liter) in the drinking water reduces their fertility. The cause of this reduction, expressed as a decrease in the number of mated females showing inplantations, was investigated, using an in vivo fertilization method. It was found that spermatozoa from lead-exposed males had a significantly lower ability to fertilize mouse eggs than those from unexposed males. Preimplantation embryos, isolated from uterine horns of mice mated with lead-exposed males. Preimplantation embryos, isolated from uterine horns of mice mated with lead-exposed males, were examined. No morphologically abnormal embryos were found. However, when cultured in vitro over the implantation period, blastocysts of the group mated with lead-exposed males showed an increased frequency of delayed hatching from the zona pellucida or an inability to hatch. Among blastocysts from this group a decreased frequency of inner cell mass development was also found.

  4. Kisspeptin-54 triggers egg maturation in women undergoing in vitro fertilization

    PubMed Central

    Jayasena, Channa N.; Abbara, Ali; Comninos, Alexander N.; Nijher, Gurjinder M.K.; Christopoulos, Georgios; Narayanaswamy, Shakunthala; Izzi-Engbeaya, Chioma; Sridharan, Mathini; Mason, Alexina J.; Warwick, Jane; Ashby, Deborah; Ghatei, Mohammad A.; Bloom, Stephen R.; Carby, Anna; Trew, Geoffrey H.; Dhillo, Waljit S.

    2014-01-01

    BACKGROUND. Patients with mutations that inactivate kisspeptin signaling are infertile. Kisspeptin-54, the major circulating isoform of kisspeptin in humans, potently stimulates reproductive hormone secretion in humans. Animal studies suggest that kisspeptin is involved in generation of the luteinizing hormone surge, which is required for ovulation; therefore, we hypothesized that kisspeptin-54 could be used to trigger egg maturation in women undergoing in vitro fertilization therapy. METHODS. Following superovulation with recombinant follicle-stimulating hormone and administration of gonadotropin-releasing hormone antagonist to prevent premature ovulation, 53 women were administered a single subcutaneous injection of kisspeptin-54 (1.6 nmol/kg, n = 2; 3.2 nmol/kg, n = 3; 6.4 nmol/kg, n = 24; 12.8 nmol/kg, n = 24) to induce a luteinizing hormone surge and egg maturation. Eggs were retrieved transvaginally 36 hours after kisspeptin injection, assessed for maturation (primary outcome), and fertilized by intracytoplasmic sperm injection with subsequent transfer of one or two embryos. RESULTS. Egg maturation was observed in response to each tested dose of kisspeptin-54, and the mean number of mature eggs per patient generally increased in a dose-dependent manner. Fertilization of eggs and transfer of embryos to the uterus occurred in 92% (49/53) of kisspeptin-54–treated patients. Biochemical and clinical pregnancy rates were 40% (21/53) and 23% (12/53), respectively. CONCLUSION. This study demonstrates that a single injection of kisspeptin-54 can induce egg maturation in women with subfertility undergoing in vitro fertilization therapy. Subsequent fertilization of eggs matured following kisspeptin-54 administration and transfer of resulting embryos can lead to successful human pregnancy. TRIAL REGISTRATION. ClinicalTrials.gov NCT01667406. FUNDING. Medical Research Council, Wellcome Trust, and National Institute for Health Research. PMID:25036713

  5. Effect of high and low antral follicle count in pubertal beef heifers on in vitro fertilization (IVF)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pubertal heifers can be classified between those with high (= 25) and low (= 15) antral follicle counts (AFC). The objective of this study was to determine oocyte development and maturation (e.g., fertility) in an in vitro fertilization (IVF) system for high and low AFC heifers. From a pool of 120...

  6. The science behind 25 years of ovarian stimulation for in vitro fertilization.

    PubMed

    Macklon, Nick S; Stouffer, Richard L; Giudice, Linda C; Fauser, Bart C J M

    2006-04-01

    To allow selection of embryos for transfer after in vitro fertilization, ovarian stimulation is usually carried out with exogenous gonadotropins. To compensate for changes induced by stimulation, GnRH analog cotreatment, oral contraceptive pretreatment, late follicular phase human chorionic gonadotropin, and luteal phase progesterone supplementation are usually added. These approaches render ovarian stimulation complex and costly. The stimulation of multiple follicular development disrupts the physiology of follicular development, with consequences for the oocyte, embryo, and endometrium. In recent years, recombinant gonadotropin preparations have become available, and novel stimulation protocols with less detrimental effects have been developed. In this article, the scientific background to current approaches to ovarian stimulation for in vitro fertilization is reviewed. After a brief discussion of the relevant aspect of ovarian physiology, the development, application, and consequences of ovarian stimulation strategies are reviewed in detail.

  7. Lasers in the in-vitro fertilization laboratory

    NASA Astrophysics Data System (ADS)

    Tadir, Yona; Neev, Joseph; Berns, Michael W.

    1993-05-01

    Laser beams are routinely used in the clinical practice of assisted reproduction. The main applications are in laparoscopic and hysteroscopic surgery. The potential applications of laser microbeams as a tool for gamete manipulations are studied and basic concepts are discussed.

  8. [Gamete donation in an in vitro fertilization program].

    PubMed

    Zhang, L Z

    1993-12-01

    Two cases of gamete donation in an IVF program were reported in this paper. In the first case the wife with recurrent abortions was found to have chromosomal abnormality, her karyotype was 45,XX,-14,-14 + rob (14 Q 14 Q). She became pregnant after IVF-ET with donated ova, and was delivered of a normal male baby on June 12, 1992. The baby's karyotype was normal. The blood group of both the husband and wife was O, while that of the baby was B. The baby suffered from icterus neonatorum as a result of ABO incompatibility but recovered quickly after treatment. Further DNA finger print analysis of the husband, wife and baby, combined with the history of egg donation IVF, proved the relationship of the parents with the baby. In the second case the husband's karyotype was 46,XY, t(4; 9) (4Q+; 9Q-). The wife had also history of recurrent abortions. Since 1989, the wife received sperm donation IUI 5 times in another hospital without success. IVF-ET with donated sperms was performed in November 1991 resulting in clinical pregnancy. She was delivered of a pair of twins, one male and one female on July 20, 1992. The possibilities of different types of gamete formation during meiosis in carriers of chromosomal balanced translocation were briefly discussed. Donation of oocytes could be realized only in an IVF program, which is also the only way to propagate for the family. However, IVF with donated sperms is indicated in women with blocked tubes besides the male factor or after repeated failures of other assisted reproductive technologies. PMID:8137642

  9. Failure of in vitro fertilization and embryo replacement following infection with Chlamydia trachomatis.

    PubMed

    Rowland, G F; Forsey, T; Moss, T R; Steptoe, P C; Hewitt, J; Darougar, S

    1985-09-01

    Antibodies to Chlamydia trachomatis were detected in 54 (47.4%) of 114 infertile women attending Bourn Hall Clinic. Antibodies were solely of the IgG class and mainly of a low titer, suggesting past infection. Antibodies were found in significantly more patients with tubal damage (54.4%) than in women whose infertility was due to other causes (16.6%). Seventy-two women completed in vitro fertilization, with 52 having three embryos replaced. We found that this treatment offered the optimum chance of a pregnancy being established, and 20 (38.5%) of these women became pregnant. Antibodies to C. trachomatis were present in only six (30.0%) of the women becoming pregnant, whereas antibodies were found in 21 (65.6%) of those who failed to become pregnant. Thus past infection with C. trachomatis halved the success rate of in vitro fertilization in these patients. The implications of these findings are relevant to all aspects of infertility from prevention to in vitro fertilization treatment.

  10. The limitations of in vitro fertilization from males with severe oligospermia and abnormal sperm morphology.

    PubMed

    Yovich, J L; Stanger, J D

    1984-09-01

    Thirty-one patients whose infertility was attributed to oligospermia were included for treatment by in vitro fertilization and embryo transfer. Three subgroups were defined: severe oligospermia (less than or equal to 5 million motile sperm/ml), moderate oligospermia (6 to less than 12 million motile sperm/ml), and abnormal sperm morphology (greater than 60% atypical). The fertilization rates were compared to those of a normospermic group managed concurrently. A modified overlay technique of sperm preparation is described for oligospermic samples so that the number of motile spermatozoa inseminated into each tube or culture dish containing a mature preovulatory oocyte was similar in each category, within the range 0.5 to 2 X 10(5)/ml. Significantly fewer oocytes were fertilized in the severe oligospermic group (P less than 0.001), suggesting a reduced capacity for fertilization by spermatozoa from severely oligospermic males. The fertilization rate of oocytes was normal in the moderate oligospermic group and those with abnormal morphology, although in the latter there was a significant delay noted in reaching the pronuclear stage (P less than 0.001), and the embryos were at a less advanced stage of cleavage at the time of transfer (0.001 less than P less than 0.01). Pregnancies were achieved in both the severe and the moderate oligospermic groups, with healthy infants delivered from each.

  11. An Antioxidant Davallialactone from Phellinus baumii Enhances Sperm Penetration on In Vitro Fertilization of Pigs.

    PubMed

    Yi, Young-Joo; Lee, In-Kyoung; Lee, Sang-Myeong; Yun, Bong-Sik

    2016-03-01

    Davallialactone (DAVA) is a hispidin analogue derived from the medicinal fungus Phellinus baumii. We examined the effect of DAVA on in vitro fertilization (IVF) of pigs. Boar spermatozoa were incubated in fertilization medium with varying concentrations of DAVA, then sperm motility and reactive oxygen species (ROS) level were evaluated. Higher sperm motility was found following the addition of 0.5 or 1 µM DAVA after incubation than addition of other concentrations or controls. ROS level decreased significantly with the addition of DAVA. The rate of normal fertilization was higher in the presence of 1 µM DAVA (65.1%) than were those of other concentrations or controls (45.4~59.4%), and the highest total fertilization rate (mono- and polyspermic oocytes) was observed at 1 µM DAVA (83%). In conclusion, addition of DAVA to fertilization medium improved sperm motility, and reduced ROS level so as to potentially improve sperm-oocyte binding in IVF, suggesting the potential of a compound isolated from mushrooms in assisted reproductive technology for humans and animals. PMID:27103855

  12. An Antioxidant Davallialactone from Phellinus baumii Enhances Sperm Penetration on In Vitro Fertilization of Pigs

    PubMed Central

    Lee, In-Kyoung; Lee, Sang-Myeong

    2016-01-01

    Davallialactone (DAVA) is a hispidin analogue derived from the medicinal fungus Phellinus baumii. We examined the effect of DAVA on in vitro fertilization (IVF) of pigs. Boar spermatozoa were incubated in fertilization medium with varying concentrations of DAVA, then sperm motility and reactive oxygen species (ROS) level were evaluated. Higher sperm motility was found following the addition of 0.5 or 1 µM DAVA after incubation than addition of other concentrations or controls. ROS level decreased significantly with the addition of DAVA. The rate of normal fertilization was higher in the presence of 1 µM DAVA (65.1%) than were those of other concentrations or controls (45.4~59.4%), and the highest total fertilization rate (mono- and polyspermic oocytes) was observed at 1 µM DAVA (83%). In conclusion, addition of DAVA to fertilization medium improved sperm motility, and reduced ROS level so as to potentially improve sperm-oocyte binding in IVF, suggesting the potential of a compound isolated from mushrooms in assisted reproductive technology for humans and animals. PMID:27103855

  13. Influences of prolactin upon spermatogenesis and spermatozoa during in vitro fertilization in mice.

    PubMed

    Mori, C; Hashimoto, H; Hoshino, K; Fukuda, A; Noda, Y; Mori, T

    1988-04-01

    Hyperprolactinemia, induced by pituitary isografts for 20 weeks in male mice and confirmed by radioimmunoassay using anti-mouse prolactin serum, did not impair spermatogenesis in the testis and maturing processes of spermatozoa in the epididymis. Incubation of freshly obtained epididymal spermatozoa for 90 min in culture media containing various levels of mouse prolactin did not yield any adverse effects on percentage motility rates of epididymal spermatozoa. When the level of mouse prolactin in the preincubation medium for epididymal spermatozoa was 100 ng/ml, the rate of fertilization by these preincubated spermatozoa in the subsequent in vitro fertilization experiment was significantly lowered compared with that observed in controls. However, when the level of prolactin in preincubation media was 10 ng/ml, no significant reduction in the rate of fertilization occurred. The present experiments seem to indicate the existence of some differences in the effects of prolactin on male germ cells until they reach the tail of the epididymis and on the processes of capacitation and/or fertilization by epididymal spermatozoa after they leave the epididymis.

  14. Emotional factors and the in vitro fertilization and embryo transfer process.

    PubMed

    Mahlstedt, P P; Macduff, S; Bernstein, J

    1987-08-01

    In vitro fertilization is nearly always a treatment of a last resort. This fact, along with the treatment's multiplicity of procedures and intensity, place unique emotional demands on patients. The goal of this research was to describe both the acknowledged emotional state of patients at the time they began the in vitro fertilization and embryo transfer (IVF-ET) process and the emotional experience of the actual procedures themselves. The findings can be used to develop strategies for providing emotional support. Data were collected from self-administered questionnaires returned by 94 IVF-ET patients in three Houston programs during a 6-month period. At the time of the IVF procedure, 77% of the population reported that infertility was still a painful concern, not something with which they had learned to live. The loss of control, seen by most patients as infertility's most stressful dimension, left them vulnerable to the intense stresses of in vitro fertilization, less able to handle its multiple demands. Thus for many, the IVF-ET procedures were like an emotional roller coaster on which they experienced a wide range of feelings during a brief period of time. Not surprisingly, emotional strain was a major consideration influencing the decision whether or not to repeat IVF. Patients indicated specific services which the staff could provide to reduce the stress of the procedures.

  15. School functioning in 8- to 18-year-old children born after in vitro fertilization.

    PubMed

    Wagenaar, Karin; Ceelen, Manon; van Weissenbruch, Mirjam M; Knol, Dirk L; Delemarre-van de Waal, Henriette A; Huisman, Jaap

    2008-11-01

    The aim of this study was to examine the school functioning of 8- to 18-year-old children born after in vitro fertilization (IVF). We compared 233 children born after IVF to 233 matched control children born spontaneously from parents with fertility problems on measures of education level, general cognitive ability, school performance (need for extra help, repeating a grade, special education), and rates of learning and developmental disorders. No differences were found between IVF and control children on these measures of school functioning. More than 60% of adolescents at secondary school attended high academic levels (with access to high school or university). We conclude that children and adolescents born after IVF show good academic achievement and general cognitive ability. They do not experience any more educational limitations than the naturally conceived children and adolescents of the control group. The tendency of reassuring school functioning already found in younger IVF children has been shown to continue at secondary school age.

  16. Nitric Oxide Synthase (NOS) Inhibition during Porcine In Vitro Maturation Modifies Oocyte Protein S-Nitrosylation and In Vitro Fertilization

    PubMed Central

    Romero-Aguirregomezcorta, Jon; Santa, Ángela Patricia; García-Vázquez, Francisco Alberto; Coy, Pilar; Matás, Carmen

    2014-01-01

    Nitric oxide (NO) is a molecule involved in many reproductive processes. Its importance during oocyte in vitro maturation (IVM) has been demonstrated in various species although sometimes with contradictory results. The objective of this study was to determine the effect of NO during IVM of cumulus oocyte complexes and its subsequent impact on gamete interaction in porcine species. For this purpose, IVM media were supplemented with three NOS inhibitors: NG-nitro-L-arginine methyl ester (L-NAME), NG-monomethyl-L-arginine (L-NMMA) and aminoguanidine (AG). A NO donor, S-nitrosoglutathione (GSNO), was also used. The effects on the cumulus cell expansion, meiotic resumption, zona pellucida digestion time (ZPdt) and, finally, on in vitro fertilization (IVF) parameters were evaluated. The oocyte S-nitrosoproteins were also studied by in situ nitrosylation. The results showed that after 42 h of IVM, AG, L-NAME and L-NMMA had an inhibitory effect on cumulus cell expansion. Meiotic resumption was suppressed only when AG was added, with 78.7% of the oocytes arrested at the germinal vesicle state (P<0.05). Supplementation of the IVM medium with NOS inhibitors or NO donor did not enhance the efficiency of IVF, but revealed the importance of NO in maturation and subsequent fertilization. Furthermore, protein S-nitrosylation is reported for the first time as a pathway through which NO exerts its effect on porcine IVM; therefore, it would be important to determine which proteins are nitrosylated in the oocyte and their functions, in order to throw light on the mechanism of action of NO in oocyte maturation and subsequent fertilization. PMID:25542028

  17. Validation of a heterologous fertilization assay and comparison of fertilization rates of equine oocytes using in vitro fertilization, perivitelline, and intracytoplasmic sperm injections.

    PubMed

    Sessions-Bresnahan, D R; Graham, J K; Carnevale, E M

    2014-07-15

    IVF in horses is rarely successful. One reason for this could be the failure of sperm to fully capacitate or exhibit hyperactive motility. We hypothesized that the zona pellucida (ZP) of equine oocytes prevents fertilization in vitro, and bypassing the ZP would increase fertilization rates. Limited availability of equine oocytes for research has necessitated the use of heterologous oocyte binding assays using bovine oocytes. We sought to validate an assay using bovine oocytes and equine sperm and then to demonstrate that bypassing the ZP using perivitelline sperm injections (PVIs) with equine sperm capacitated with dilauroyl phosphatidylcholine would result in higher fertilization rates than standard IVF in bovine and equine oocytes. In experiment 1, bovine oocytes were used for (1) IVF with bovine sperm, (2) IVF with equine sperm, and (3) intracytoplasmic sperm injections (ICSIs) with equine sperm. Presumptive zygotes were either stained with 4',6-diamidino-2-phenylindole from 18 to 26 hours at 2-hour intervals or evaluated for cleavage at 56 hours after addition of sperm. Equine sperm fertilized bovine oocytes; however, pronuclei formation was delayed compared with bovine sperm after IVF. The delayed pronuclear formation was not seen after ICSI. In experiment 2, bovine oocytes were assigned to the following five groups: (1) cumulus oocyte complexes (COCs) coincubated with bovine sperm; (2) COC exposed to sucrose then coincubated with bovine sperm; (3) COC coincubated with equine sperm; (4) COC exposed to sucrose, and coincubated with equine sperm; and (5) oocytes exposed to sucrose, and 10 to 15 equine sperm injected into the perivitelline (PV) space. Equine sperm tended (P = 0.08) to fertilize more bovine oocytes when injected into the PV space than after IVF. In experiment 3, oocytes were assigned to the following four groups: (1) IVF, equine, and bovine COC coincubated with equine sperm; (2) PVI of equine and bovine oocytes; (3) PVI with equine oocytes

  18. Urinary bisphenol A concentrations and association with in vitro fertilization outcomes among women from a fertility clinic

    PubMed Central

    Mínguez-Alarcón, Lidia; Gaskins, Audrey J.; Chiu, Yu-Han; Williams, Paige L.; Ehrlich, Shelley; Chavarro, Jorge E.; Petrozza, John C.; Ford, Jennifer B.; Calafat, Antonia M.; Hauser, Russ

    2015-01-01

    STUDY QUESTION Are urinary BPA concentrations associated with in vitro fertilization (IVF) outcomes among women attending an academic fertility center? SUMMARY ANSWER Urinary BPA concentrations were not associated with adverse reproductive and pregnancy outcomes among women from a fertility clinic. WHAT IS KNOWN ALREADY Bisphenol A (BPA), an endocrine disruptor, is detected in the urine of most Americans. Although animal studies have demonstrated that BPA reduces female fertility through effects on the ovarian follicle and uterus, data from human populations are scarce and equivocal. STUDY DESIGN, SIZE AND DURATION This prospective cohort study between 2004 and 2012 at the Massachusetts General Hospital Fertility Center included 256 women (n = 375 IVF cycles) who provided up to two urine samples prior to oocyte retrieval (total N = 673). PARTICIPANTS/MATERIALS, SETTINGS, METHODS Study participants were women enrolled in the Environment and Reproductive Health (EARTH) Study. Intermediate and clinical end-points of IVF treatments were abstracted from electronic medical records. We used generalized linear mixed models with random intercepts to evaluate the association between urinary BPA concentrations and IVF outcomes adjusted by age, race, body mass index, smoking status and infertility diagnosis. MAIN RESULTS AND THE ROLE OF CHANCE The specific gravity-adjusted geometric mean of BPA was 1.87 µg/l, which is comparable to that for female participants in the National Health and Nutrition Examination Survey, 2011–2012. Urinary BPA concentrations were not associated with endometrial wall thickness, peak estradiol levels, proportion of high quality embryos or fertilization rates. Furthermore, there were no associations between urinary BPA concentrations and implantation, clinical pregnancy or live birth rates per initiated cycle or per embryo transfer. Although we did not find any associations between urinary BPA concentrations and IVF outcomes, the relation between

  19. Development of an in vitro index to characterize fertilizing capacity of boar ejaculates.

    PubMed

    Schulze, M; Ruediger, K; Mueller, K; Jung, M; Well, C; Reissmann, M

    2013-07-01

    The aim of this research was the selection of spermatozoa parameters related to boar fertility performance and their combination into an in vitro index. A first set (data set 1) of 36 Pietrain boars with 138 ejaculates from two seasons with 5083 single-sire inseminations from 34 farms was used to determine correlations between in vitro sperm quality parameters and fertility performance. 2970 ejaculates representing a second set (data set 2) served calculation of seasonal and age effects on semen quality. Morphological spermatozoa parameters were estimated manually with a phase contrast microscope on the day of semen collection, whereas mitochondrial activity and viability were analyzed by double-staining with rhodamine123/propidium iodide on day 2 of semen storage using flow cytometry. Sperm motility was tested on day 7 by thermoresistance (TRT) after 30min (TRT1) and 300min (TRT2) incubation at 38̊C using computer-assisted semen analysis (CASA). Correlations revealed four independent sperm quality parameters qualifying as relevant predictors of boar fertility: (i) percentage of spermatozoa with proximal cytoplasmic droplets, (ii) percentage of spermatozoa with active mitochondria, (iii) beat cross frequency of progressively motile spermatozoa in TRT1, and (iv) oscillation measure of the actual path of progressively motile spermatozoa in TRT2. There were no significant effects of sperm concentration, ejaculate volume, and total number of sperm cells per ejaculate on litter size (LS) and on pregnancy rate (PR). Our findings suggest the usefulness of sperm quality parameters based on adjusted range of methods and enable the construction of an in vitro index as a means to predicting boar fertility. PMID:23773327

  20. Protective effects of cornus mas extract on in vitro fertilization potential in methotrexate treated male mice

    PubMed Central

    Zarei, Leila; Shahrooz, Rasoul; Sadrkhanlou, Rajabali; Malekinejad, Hassan; Ahmadi, Abbas; Bakhtiary, Zahra

    2015-01-01

    Current study was aimed to evaluating protective effects of cornus mas fruit extract (CMFE) in mice treated with methotrexate (MTX). For this purpose, 48 young mature male mice were divided into 6 groups. Control group received only normal saline (0.1 mL per day, intraperitoneally), and the second group was administered MTX (20 mg kg-1 per week, intraperitoneally). The third, fourth and fifth groups received MTX daily oral doses of 250, 500 and 1000 mg kg-1 CMFE as well as MTX. The sixth group was only given CMFE with a dose of 1000 mg kg-1 per day, orally, for 35 days. Then, the animals were anesthetically euthanized and the sperms were separated from epididymis. DNA damage level, the amount of malondialdehyde (MDA) as well as in vitro fertility was evaluated. The number of sperms with damaged DNA and MDA level in MTX-treated group showed a significant increase compared to control group (p < 0.05). In groups receiving CMFE along with MTX, DNA damage level and MDA amount suggested a decrease in comparison with MTX group (p < 0.05). Also, in vitro fertilization and embryonic development in MTX-treated group was significantly lower than the control group, and the level of embryonic arresting was higher (p < 0.05). In groups which received CMFE along with MTX, in vitro fertility and embryonic development was higher than MTX group (p < 0.05) and the arrested embryos showed a decrease. This study suggested that cornus mas is able to ameliorate the side effects of MTX. PMID:25992252

  1. In vitro fertilization for endometriosis-associated infertility.

    PubMed

    Polat, Mehtap; Yaralı, İrem; Boynukalın, Kübra; Yaralı, Hakan

    2015-08-01

    Endometriosis is an enigmatic disease affecting 10-15% of reproductive aged women and is encountered in 25-35% of women suffering from infertility. IVF is an effective tool to overcome endometriosis-associated infertility when expectant management or surgery fails. Direct IVF should be envisioned if the female age is greater than 38 year and infertility is long lasting. Likewise, semen characteristics or tubal status that is incompatible with natural conception mandates going straight to IVF. IVF, not only bypasses the distortion of pelvic anatomy associated with advanced stage endometriosis, but also removes gametes from a hostile peritoneal environment. In this article, we address the impact, if any, of endometriosis and endometriomason IVF outcome, whether surgical treatment of early-stage disease, endometriomas or deep infiltrating endometriosis would enhance pregnancy rates in IVF, which protocol to employ for controlled ovarian hyperstimulation for IVF and finally the impact, if any, of controlled ovarian hyperstimulation for IVF on progression of endometriosis.

  2. Reductive dechlorination reduces negative impact of PCBs on in vitro fertilization

    SciTech Connect

    Chou, K.; Mousa, M.; Quensen, J.

    1995-12-31

    The variety of toxic effects ascribed to commercial PCB mixtures is likely because the individual congeners differ in their modes of toxicity and potencies. Similarly, a change in the congener composition of a PCB mixture due to reductive dechlorination by anaerobic microorganisms might be expected to alter the toxicological properties of that PCB mixture. In this study, Aroclors 1,242 and 1,254 were reductively dechlorinated using cultures of microorganisms from two different sites. The resulting dechlorination products and Aroclors were then tested for their effects on in vitro fertilization using mouse gametes. In general, percent fertilization increased with the extent of dechlorination. For example, fertilization in treatments with 10 {micro}g/ml of either Aroclor was only 40% of that in controls without Aroclor, but equal total molar concentrations of the dechlorination products produced by River Raisin microorganisms had no significant effect on fertilization. By this measure, therefore, the microbial dechlorination of PCBs appears to have a beneficial effect in reducing reproductive toxicity.

  3. The efficiency of cryopreserved semen versus fresh semen for in vitro fertilization/embryo transfer.

    PubMed

    Yavetz, H; Lessing, J B; Niv, Y; Amit, A; Barak, Y; Yovel, I; David, M P; Peyser, M R; Yogev, L; Homonnai, Z

    1991-06-01

    The efficiency of cryopreserved donor semen versus fresh donor semen in an in vitro fertilization/embryo transfer programme was evaluated. Thirty-nine in vitro fertilization/embryo transfer cycles were performed using fresh donor semen (group A) and 74 cycles were carried out using cryopreserved semen (group B). All patients underwent a uniform controlled ovarian hyperstimulation using high doses of human menopausal gonadotropins. Oocytes were retrieved transvaginally under ultrasound imaging. Semen (fresh or frozen-thawed) were prepared for insemination by a washing technique. Each ovum was inseminated with 500,000-600,000 motile spermatozoa. No significant difference was noted between the two groups regarding female age, duration of infertility, and number of ova retrieved per aspiration. Even though the fertilization rate in group B was significantly lower than in group A (55.5 +/- 3.8 vs 70.4 +/- 3.5, P = 0.008), pregnancy rates per embryo transfer were similar--39.3 and 38.5%, respectively.

  4. In vitro maturation, fertilization and embryo development after ultrarapid freezing of immature human oocytes.

    PubMed

    Wu, J; Zhang, L; Wang, X

    2001-03-01

    The purpose of this study was to determine the rates of maturation, fertilization and embryo development of ultrarapidly frozen immature oocytes (immature cumulus-oocyte complexes; COCs) obtained from antral follicles in ovaries of patients with chocolate ovarian cysts. The COCs were cryopreserved by a vitrification method using 5.5 mol ethylene glycol l (-1) plus 1.0 mol sucrose l (-1) in Dulbecco's PBS (DPBS). The survival, maturation and fertilization rates, and the percentage of embryos developing to the two-cell stage were 59, 64, 70 and 71%, respectively. No significant differences were noted in the rates of maturation, fertilization and embryo development between control and cryopreserved oocytes. Two embryos that developed from cryopreserved oocytes of the oocyte donor programme were selected for transfer into the uterus of a recipient with premature ovarian failure, after the recipient had received steroid replacement. A biochemical pregnancy occurred in the recipient after embryo transfer. These results indicate that immature oocytes can survive after cryopreservation and subsequently can be cultured to mature oocytes that are capable of undergoing fertilization in vitro and developing into embryos.

  5. Female fertility preservation strategies: cryopreservation and ovarian tissue in vitro culture, current state of the art and future perspectives.

    PubMed

    Filatov, M A; Khramova, Y V; Kiseleva, M V; Malinova, I V; Komarova, E V; Semenova, M L

    2016-10-01

    In the present review, the main strategies of female fertility preservation are covered. Procedures of fertility preservation are necessary for women who suffer from diseases whose treatment requires the use of aggressive therapies, such as chemotherapy and radiotherapy. These kinds of therapy negatively influence the health of gametes and their progenitors. The most commonly used method of female fertility preservation is ovarian tissue cryopreservation, followed by the retransplantation of thawed tissue. Another approach to female fertility preservation that has been actively developed lately is the ovarian tissue in vitro culture. The principal methods, advantages and drawbacks of these two strategies are discussed in this article.

  6. Sex-sorting of spermatozoa affects developmental competence of in vitro fertilized oocytes in a bull-dependent manner

    PubMed Central

    INABA, Yasushi; ABE, Reika; GESHI, Masaya; MATOBA, Satoko; NAGAI, Takashi; SOMFAI, Tamás

    2016-01-01

    The aim of the present study was to clarify if flow-cytometric sex-sorting of bovine sperm affected in vitro blastocyst production in different bulls, either in terms of its ability to fertilize the oocyte or by interfering with post-fertilization embryo development. We performed in vitro fertilization (IVF) using both commercially available frozen-thawed X-sorted and non-sorted sperm of 4 Holstein bulls at 3 concentrations (1 × 106, 2 × 106, and 5 × 106 sperm/ml). When fertilization rates were compared, a variation in fertilization rates among different sperm concentrations was detected in 2 bulls, with similar results for X-sorted and non-sorted sperm. However, we found no evidence that the fertilization rates were affected by the sorting process. To investigate effects on embryo development, we determined the optimum sperm concentration for IVF in each bull, which resulted in similar fertilization rates among bulls. We next performed IVF using both X-sorted and non-sorted sperm of the 4 bulls at their optimum sperm concentration and compared in vitro embryo development. Cleavage rates with X-sorted sperm were similar to their non-sorted counterparts. However, significantly reduced blastocyst development was associated with the use of X-sorted sperm in one bull, whereas in the other three bulls, blastocyst development after IVF with X-sorted and non-sorted sperm was similar. In conclusion, in our system, X-sorting affects in vitro blastocyst production by reducing the developmental competence of fertilized oocytes rather than affecting the fertilization ability of the sperm. However, the occurrence of this phenomenon varies among bulls. PMID:27301424

  7. Quality management systems for your in vitro fertilization clinic's laboratory: Why bother?

    PubMed Central

    Olofsson, Jan I; Banker, Manish R; Sjoblom, Late Peter

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF) clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART) laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services. This review contains the proceedings a quality management session at the Indian Fertility Experts Meet (IFEM) 2010 and focuses on the creation of a patient-oriented best-in-class IVF laboratory. PMID:23869142

  8. Outcome of patients with one ovary in an in vitro fertilization program.

    PubMed

    Lam, S Y; McKenna, M; McBain, J C; Baker, H W; Johnston, W I

    1987-12-01

    To determine whether the absence of one ovary would influence adversely the outcome of in vitro fertilization, the results of 60 laparoscopic oocyte retrieval procedures in 34 single-ovary patients were compared with the outcome for all laparoscopic retrieval cycles during the same period (January 1984-August 1986) in patients with tubal infertility and two accessible ovaries (559 cycles in 335 patients). The median age was younger in the one-ovary group (31 vs 32) (P less than 0.05). The group with two accessible ovaries had significantly more follicles aspirated (median, 7 vs 5; P less than 0.001) and more oocytes obtained (median, 5 vs 4; P less than 0.001) per retrieval compared with the single-ovary group. However, the single-ovary group had a higher percentage fertilization (69 vs 62%; P less than 0.05) so that the number of embryos transferred per transfer patient (median, 3 vs 3) and pregnancy rate per cycle (9.7 vs 15.6%) did not differ significantly between groups. We conclude that patients with a single ovary have a compensatory increase in the ability to produce oocytes which may have greater potential for fertilization and subsequent development.

  9. The Effect of Complementary and Alternative Medicine on Subfertile Women with In Vitro Fertilization

    PubMed Central

    Zhang, Yuehui; Fu, Yiman; Han, Fengjuan; Kuang, Hongying; Hu, Min; Wu, Xiaoke

    2014-01-01

    About 10–15% of couples have difficulty conceiving at some point in their reproductive lives and thus have to seek specialist fertility care. One of the most commonly used treatment options is in vitro fertilization (IVF) and its related expansions. Despite many recent technological advances, the average IVF live birth rate per single initiated cycle is still only 30%. Consequently, there is a need to find new therapies to promote the efficiency of the procedure. Many patients have turned to complementary and alternative medical (CAM) treatments as an adjuvant therapy to improve their chances of success when they undergo IVF treatment. At present, several CAM methods have been used in infertile couples with IVF, which has achieved obvious effects. However, biologically plausible mechanisms of the action of CAM for IVF have not been systematically reviewed. This review briefly summarizes the current progress of the impact of CAM on the outcomes of IVF and introduces the mechanisms. PMID:24527047

  10. Scheduling cycles with gonadotropin-releasing hormone antagonist protocol in in vitro fertilization: Is there a scope in batch in vitro fertilization?

    PubMed Central

    Gutgutia, Rohit; Rao, Sameer; Garcia-Velasco, Juan; Basu, Susmita

    2014-01-01

    In India, a practice of “Batch in vitro fertilization (IVF)” has evolved in many infertility centers in an effort to align infertility management with logistics. A “Batch IVF” is an approach where the menstrual cycles of multiple women are programmed, such that they can undergo all the processes; from stimulation until embryo transfer about the same time. In “Batch IVF”, the day for initiating stimulation is calculated retrospectively from the day the visiting embryologist is available at the clinic (day of ovum pick-up). Aligning the cycles of multiple women with steroids followed by down regulation with long gonadotropin-releasing hormone agonist (GnRH-A) is one of the currently employed methods for batching. There is sufficient evidence on scheduling cycles with steroids in GnRH-An protocol without compromising on the outcome. The objective of this paper is to provide evidence-based clinical concept on scheduling cycles in “Batch IVF” setup with GnRH-An protocol through literature review. PMID:25624658

  11. Scheduling cycles with gonadotropin-releasing hormone antagonist protocol in in vitro fertilization: Is there a scope in batch in vitro fertilization?

    PubMed

    Gutgutia, Rohit; Rao, Sameer; Garcia-Velasco, Juan; Basu, Susmita

    2014-01-01

    In India, a practice of "Batch in vitro fertilization (IVF)" has evolved in many infertility centers in an effort to align infertility management with logistics. A "Batch IVF" is an approach where the menstrual cycles of multiple women are programmed, such that they can undergo all the processes; from stimulation until embryo transfer about the same time. In "Batch IVF", the day for initiating stimulation is calculated retrospectively from the day the visiting embryologist is available at the clinic (day of ovum pick-up). Aligning the cycles of multiple women with steroids followed by down regulation with long gonadotropin-releasing hormone agonist (GnRH-A) is one of the currently employed methods for batching. There is sufficient evidence on scheduling cycles with steroids in GnRH-An protocol without compromising on the outcome. The objective of this paper is to provide evidence-based clinical concept on scheduling cycles in "Batch IVF" setup with GnRH-An protocol through literature review. PMID:25624658

  12. In vitro fertilization of oocytes from polyovular follicles in mouse ovaries exposed neonatally to diethylstilbestrol.

    PubMed

    Iguchi, T; Kamiya, K; Uesugi, Y; Sayama, K; Takasugi, N

    1991-01-01

    In 35-day-old female ICR/JCL mice given 5 daily injections of 1 microgram diethylstilbestrol (DES) from the day of birth, a significantly higher incidence of polyovular follicles was found in the ovaries than in those of age-matched control mice. Gap junctions of granulosa cells of mature follicles in neonatally DES-exposed mice were larger than those of the controls. When stimulated by gonadotropins (PMSG and hCG) and caged with males, the number of tubal embryos in DES-exposed mice was less, but the rate of fertilization and development was not different compared to the controls. Division of oocytes collected from the ovaries of 40-day-old DES-exposed and control mice after stimulation of gonadotropins was examined 24 to 72 h after in vitro insemination to ascertain whether fertilization had occurred in oocytes from polyovular follicles. Seventy-seven % of oocytes from uniovular follicles of control mice developed up to 8-cell stage embryos following in vitro insemination; 66% of those from similar follicles of DES-exposed mice developed into the same stage. By contrast, only 47% of oocytes from polyovular follicles of DES-exposed mice showed the division up to 8-cell stage 72 h after insemination, indicating a significantly lower fertilization rate compared to the oocytes from uniovular follicles of control and DES-exposed mice. Without insemination, oocytes taken from the same pools in these experiments never divided during the period of manipulation and incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Transvaginal recovery of oocytes for in vitro fertilization using vaginal ultrasound.

    PubMed

    Lenz, S; Leeton, J; Renou, P

    1987-02-01

    Vaginal recovery of oocytes for in vitro fertilization (IVF) was performed in 75 patients using a vaginal puncture transducer. Follicles were aspirated and flushed four times using a controlled vacuum. Oocytes were collected in 74 patients and an average of 5.1 oocytes was collected per patient. An average of 8.8 follicles was aspirated per procedure, giving an oocyte recovery rate of 58% per follicle. Pregnancy was obtained in seven patients (9.3%). The only complication was bleeding from the vagina of an estimated 40 ml in one case.

  14. Complete in vitro generation of fertile oocytes from mouse primordial germ cells

    PubMed Central

    Morohaku, Kanako; Tanimoto, Ren; Sasaki, Keisuke; Kawahara-Miki, Ryouka; Kono, Tomohiro; Hayashi, Katsuhiko; Hirao, Yuji; Obata, Yayoi

    2016-01-01

    Reconstituting gametogenesis in vitro is a key goal for reproductive biology and regenerative medicine. Successful in vitro reconstitution of primordial germ cells and spermatogenesis has recently had a significant effect in the field. However, recapitulation of oogenesis in vitro remains unachieved. Here we demonstrate the first reconstitution, to our knowledge, of the entire process of mammalian oogenesis in vitro from primordial germ cells, using an estrogen-receptor antagonist that promotes normal follicle formation, which in turn is crucial for supporting oocyte growth. The fundamental events in oogenesis (i.e., meiosis, oocyte growth, and genomic imprinting) were reproduced in the culture system. The most rigorous evidence of the recapitulation of oogenesis was the birth of fertile offspring, with a maximum of seven pups obtained from a cultured gonad. Moreover, cryopreserved gonads yielded functional oocytes and offspring in this culture system. Thus, our in vitro system will enable both innovative approaches for a deeper understanding of oogenesis and a new avenue to create and preserve female germ cells. PMID:27457928

  15. Complete in vitro generation of fertile oocytes from mouse primordial germ cells.

    PubMed

    Morohaku, Kanako; Tanimoto, Ren; Sasaki, Keisuke; Kawahara-Miki, Ryouka; Kono, Tomohiro; Hayashi, Katsuhiko; Hirao, Yuji; Obata, Yayoi

    2016-08-01

    Reconstituting gametogenesis in vitro is a key goal for reproductive biology and regenerative medicine. Successful in vitro reconstitution of primordial germ cells and spermatogenesis has recently had a significant effect in the field. However, recapitulation of oogenesis in vitro remains unachieved. Here we demonstrate the first reconstitution, to our knowledge, of the entire process of mammalian oogenesis in vitro from primordial germ cells, using an estrogen-receptor antagonist that promotes normal follicle formation, which in turn is crucial for supporting oocyte growth. The fundamental events in oogenesis (i.e., meiosis, oocyte growth, and genomic imprinting) were reproduced in the culture system. The most rigorous evidence of the recapitulation of oogenesis was the birth of fertile offspring, with a maximum of seven pups obtained from a cultured gonad. Moreover, cryopreserved gonads yielded functional oocytes and offspring in this culture system. Thus, our in vitro system will enable both innovative approaches for a deeper understanding of oogenesis and a new avenue to create and preserve female germ cells. PMID:27457928

  16. In vitro fertilization (IVF): a review of 3 decades of clinical innovation and technological advancement

    PubMed Central

    Wang, Jeff; Sauer, Mark V

    2006-01-01

    In vitro fertilization, popularly referred to as IVF, has captured the attention of the public since its sensational introduction in 1978. Today assisted reproductive technology is available throughout most of the civilized world, and the practice is largely different from that used during the early days. Refinements in laboratory technology and clinical practice have allowed IVF to evolve into a medical procedure that is efficient, safe, readily accessible, and relatively affordable. More than 2 million IVF children have been born to date, and it is likely that continued enhancements will widen its appeal and applicability. PMID:18360648

  17. Autophagy induction by sera from women undergoing an in vitro fertilization cycle varies with subsequent outcome.

    PubMed

    Sisti, Giovanni; Kanninen, Tomi T; Di Tommaso, Mariarosaria; Witkin, Steven S; Spandorfer, Steven D

    2016-09-01

    Autophagy maintains intracellular homeostasis during placental development and embryogenesis. We evaluated if differences in the autophagy-inducing capacity of sera from women undergoing an in vitro fertilization (IVF) cycle predicted subsequent pregnancy outcome. In this retrospective study, sera collected from 94 women at the time of intrauterine embryo implantation were incubated with peripheral blood mononuclear cells (PBMCs) from healthy donors. The PBMCs were lysed and autophagy induction measured by determination of the p62 concentration. A reduced capacity for autophagy induction was associated with defective implantation while an elevated level of autophagy was associated with ectopic pregnancy. PMID:27343871

  18. In vitro fertilization (IVF): a review of 3 decades of clinical innovation and technological advancement.

    PubMed

    Wang, Jeff; Sauer, Mark V

    2006-12-01

    In vitro fertilization, popularly referred to as IVF, has captured the attention of the public since its sensational introduction in 1978. Today assisted reproductive technology is available throughout most of the civilized world, and the practice is largely different from that used during the early days. Refinements in laboratory technology and clinical practice have allowed IVF to evolve into a medical procedure that is efficient, safe, readily accessible, and relatively affordable. More than 2 million IVF children have been born to date, and it is likely that continued enhancements will widen its appeal and applicability.

  19. The technology, law, and ethics of in vitro fertilization, gamete donation, and surrogate motherhood.

    PubMed

    Wasserman, D; Wachbroit, R

    1992-09-01

    This article examines some of the legal, ethical, and policy issues raised by the development and use of technologies for noncoital reproduction: gamete transfer and manipulation, in vitro fertilization, and zygote transfer and manipulation. After briefly describing these technologies, this article examines three closely related concerns raised by their introduction: (1) the effect of new technologies on the social understanding of parenthood and the legal regulation of the family; (2) the impact on women and children of a market in the material and services for producing children; and (3) the rights and interests involved in conflicts over the control and disposition of extrauterine embryos. PMID:1521422

  20. Use of combinations of in vitro quality assessments to predict fertility of bovine semen.

    PubMed

    Sellem, E; Broekhuijse, M L W J; Chevrier, L; Camugli, S; Schmitt, E; Schibler, L; Koenen, E P C

    2015-12-01

    Predicting in vivo fertility of bull ejaculates using in vitro-assessed semen quality criteria remains challenging for the breeding industry. New technologies such as computer-assisted semen analysis (CASA) and flow cytometry may provide accurate and objective methods to improve semen quality control. The aim of this study was to evaluate the relationship between semen quality parameters and field fertility of bull ejaculates. A total of 153 ejaculates from 19 Holstein bulls have been analyzed using CASA (postthawing semen motility and morphology) and several flow cytometric tests, including sperm DNA integrity, viability (estimated by membrane integrity), acrosomal integrity, mitochondria aerobic functionality and oxidation. Samples were analyzed both immediately after thawing and after 4 hours at 37 °C. A fertility value (FV), based on nonreturn rate at 56 days after insemination and adjusted for environment factors, was calculated for each ejaculate. Simple and multiple regressions have been used to correlate FV with CASA and flow cytometric parameters. Significant simple correlations have been observed between some parameters and FV (e.g., straight line velocity [μm/s], r(2) = -0.12; polarized mitochondria sperm (%), r(2) = 0.07), but the relation between simple parameter and FV was too week to predict the fertility. Partial least square procedure identified several mathematical models combining flow cytometer and CASA variables and had better correlations with FV (adjusted r(2) ranging between 0.24 and 0.40 [P < 0.0001], depending on the number of included variables). In conclusion, this study suggests that quality assessment of thawed bull sperm using CASA and flow cytometry may provide a reasonable prediction of bovine semen fertility. Additional work will be required to increase the prediction reliability and promote this technology in routine artificial insemination laboratory practice.

  1. In vitro culture and in vitro fertilization techniques for prairie voles (Microtus ochrogaster).

    PubMed

    Horie, Kengo; Hidema, Shizu; Hirayama, Takashi; Nishimori, Katsuhiko

    2015-08-01

    Prairie vole (Microtus ochrogaster) is a highly social animal and is a commonly used animal model for neuropsychopharmacological and psychiatric studies. To date, only a few reports on the development of transgenic prairie voles which was primarily due to the suboptimal development of assisted reproductive technology (ART) in prairie voles. Limitations in ART further hinder the development of genetically modified prairie voles such as the application of conventional gene targeting technologies using embryonic stem (ES) or induced pluripotent stem (iPS) cells to generate chimeric prairie voles. Moreover, recent advancement in genome-editing tools such as transcription activator-like effector nuclease (TALEN) and clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas technology provide an unprecedented opportunity to create gene targeting animal model and the development of ART in prairie voles is necessary for future development of novel transgenic prairie vole model. We have established efficient method for in vitro embryo culture and sperm cryopreservation with high fertilization rate. In G-1 PLUS and G-2 PLUS sequential culture condition, 81.0% (# of Blastocysts/total n) of one-cell embryos developed to blastocysts. In contrary, no embryos were developed to blastocyst stage in KSOM medium (0/total # of embryos in culture). In vitro fertilization rate using fresh and frozen-thawed sperm was 32.6% and 29.3%, respectively. This is the first report of IVF using cryopreserved prairie vole sperm. We employed mouse IVF methods in prairie voles and optimize culture conditions using human G-1/G-2 PLUS sequential culture method that resulted in high embryonic development rate. The development in vole reproductive technology will facilitate the generation of transgenic voles in the future. PMID:26071353

  2. [Life-threatening extrauterine pregnancy after in vitro fertilization and bilateral salpingectomy].

    PubMed

    Claudi, Anne; Hansen, Christian Steen; Nørgaard, Lone Nikoline

    2015-01-26

    A woman with a history of bilateral salpingectomy achieved pregnancy after in vitro fertilization. At the gestational age of six weeks she experienced fatigue, haematemesis and abdominal pain. She was hospitalized on suspicion of a peptic ulcer. Several physicians rejected the possibility of an ectopic pregnancy due to the history of salpingectomy. The patient had intraabdominal bleeding and went through emergency surgery because of a ruptured interstitial pregnancy. The purpose of this case report is to recall the risk of interstitial pregnancy in pregnant women with a history of salpingectomy.

  3. [Successful treatment of a cervical heterotopic pregnancy following an in vitro fertilization procedure].

    PubMed

    Elena, Hernán E; Elena, Alfredo F; Miola, Anselmo; Glujovsky, Demian; Sueldo, Carlos E

    2016-01-01

    A 37-year-old nulligravida infertile female had a cervical heterotopic pregnancy following an in vitro fertilization procedure. Early intervention on the 6th week of gestation with a manual vacuum aspirator reached to remove the cervical pregnancy. Ligation of the descending cervical branches of the uterine arteries and a cervical cerclage, were placed before the aspiration, for prevention of possible hemorrhage. Successful removal of the cervical pregnancy was achieved with only mild bleeding. An intrauterine pregnancy progressed to viability without complications, resulting in a vaginal delivery of a preterm live-birth at 35.4 weeks, of a male that weighted 2740 g.

  4. MicroRNA expression profiling of elongated cloned and in vitro-fertilized bovine embryos.

    PubMed

    Castro, F O; Sharbati, S; Rodríguez-Alvarez, L L; Cox, J F; Hultschig, C; Einspanier, R

    2010-01-01

    The objective of this study was to identify microRNAs (miRNAs) expressed in bovine (Bos Taurus) cloned embryos at Day 17 of development (Day 0=day of nucleus transfer or in vitro fertilization) during elongation. Day 7 bovine expanded blastocysts produced by hand made cloning (HMC) or in vitro fertilization were bulk-transferred to synchronized recipient cattle (48 HMC embryos to 10 recipients and 28 in vitro-produced embryos to four recipients). Elongated embryos were retrieved at Day 17; miRNAs were isolated and subjected to microarray screening using custom composite slides spotted with human, mouse, and rat and in silico-predicted miRNAs. An initial profile of expressed miRNAs was determined in cloned embryos and somatic donor cells; this profile changed after somatic cell nucleus transfer, identifying differentially expressed miRNAs between cloned and in vitro-produced bovine embryos. Furthermore, microarray data were validated using a miRNA-specific quantitative reverse transcription-polymerase chain reaction (qRT-PCR) approach (miR-Q). There was an 83% correlation (P=0.01) between microarray and qPCR data. Based on qRT-PCR, correct reprogramming of some miRNAs from the donor cells was confirmed in cloned bovine embryos, whereas other somatic miRNAs were not appropriately reprogrammed. Some of the miRNAs that were equally reprogrammed clustered on the same chromosomal location in the bovine genome. In conclusion, reprogramming of miRNAs seemed to occur in cloned bovine embryos. This could have profound implications for elucidating nuclear reprogramming in somatic cloning, as well as for the role of miRNAs in preimplantation mammalian development.

  5. Cytotoxic sperm antibodies and in vitro fertilization of mature oocytes: a preliminary report.

    PubMed

    Mathur, S; Mathur, R S; Holtz, G L; Tsai, C C; Rust, P F; Williamson, H O

    1987-06-01

    The fertilization rates of mature oocytes during in vitro fertilization and embryo transfer (IVF-ET) using fetal cord serum-supplemented insemination media were greater than or equal to 57% for five infertile couples without sperm antibodies (group 1). But they were less than or equal to 50% for four of nine infertile couples (group 2) with cytotoxic sperm antibodies in both partners (n = 6) or the husband alone (n = 3). Two women in group 1 were successful in achieving normal, full-term pregnancies with the delivery of normal infants (chi2 = 4.2, P less than 0.05, by chi-square analysis). One of them consistently tested negative for sperm antibodies, while her husband was previously treated with antibiotics for infection and transient sperm antibodies in the seminal plasma. Subsequently, antibody titers in the husband were in the normal range when the successful IVF-ET was performed. One woman in group 2, with antibodies to her autoimmune husband's sperm but not control sperm and with a long-standing poor postcoital test sperm motility, conceived through artificial insemination with donor sperm (AID) after failing to conceive with her husband through IVF-ET. These data suggest that the presence of cytotoxic sperm antibodies in the serum and/or secretions of both partners reduces the rates of fertilization of mature oocytes in spite of using fetal cord serum in the IVF media. Pregnancy achievement is impaired in this group.

  6. In vitro fertilization and embryo transfer (IVF/ET): an established and successful therapy for endometriosis.

    PubMed

    Oehninger, S; Acosta, A A; Kreiner, D; Muasher, S J; Jones, H W; Rosenwaks, Z

    1988-10-01

    The purpose of this report is to present a 6-year experience in the management of endometriosis with in vitro fertilization and embryo transfer (IVF/ET). We divided 136 patients who underwent 280 cycles into three groups: (1) previous history of endometriosis but normal pelvis at the time of oocyte retrieval, (2) stages I-II endometriosis (revised AFS classification), and (3) stages III-IV endometriosis. The stimulation protocols, estradiol (E2) responses, and distribution of terminal E2 patterns were similar in all groups. Group 3 had significantly fewer preovulatory and immature oocytes retrieved and fewer embryos transferred. The fertilization rate and the per cycle/per transfer pregnancy rates were similar in all groups. The miscarriage rate was higher in group 3, and the ongoing pregnancy rate per cycle was lower. Luteal phase E2 and progesterone levels were comparable in all groups. No differences were found when groups 2 and 3 were analyzed for the presence of one or two ovaries or the presence/absence of ovarian endometriosis. The overall fertilization rate, the per cycle/per transfer pregnancy rates, and the miscarriage rate were similar to those of tubal factor patients. We underscore the excellent outcome of patients with minimal or mild endometriosis in IVF/ET. We conclude that patients with moderate or severe endometriosis have a compromised reproductive potential, probably because of a reduced oocyte recovery rate and poor embryo quality.

  7. Identification of male-factor semen samples prior to insemination and in vitro fertilization.

    PubMed

    Chan, P J; Tredway, D R; Su, B C; Corselli, J; Davidson, B; Ren, S

    1991-02-01

    Semen analyses carried out as part of the clinical in vitro fertilization or intrauterine insemination protocols provide important information that determine the type of clinical treatment of the male partner and the sperm processing method. It is postulated that the sperm of male-factor patients cannot survive hypoosmotic stress conditions because of defective sperm membrane function. To test this, 0.1 ml of semen from each of 102 patients was placed in 1.0 ml of 150 mosmol/liter eosin citrate fructose solution and incubated for 30 min at 37 degrees C. The percentage viability of the sperm cells was then determined. The results indicated that patients with two or more abnormal semen parameters had a significantly lower percentage viability while in the hypoosmotic solution (40.6 +/- 4.7%), in contrast to non-male-factor patients (69.0 +/- 1.6%). Donor sperm (N = 32) serving as controls (73.3 +/- 2.1%) had a viability in hypoosmotic solution similar to that of non-male-factor patients. The data suggest that sperm of male-factor patients are less able to survive the hypoosmotic stress conditions as shown by the percentage viability in hypoosmotic solution and emphasize the importance of using less stressful sperm processing methods for in vitro fertilization or insemination in these patients.

  8. Cost and efficacy comparison of in vitro fertilization and tubal anastomosis for women after tubal ligation

    PubMed Central

    Messinger, Lauren B.; Alford, Connie E.; Csokmay, John M.; Henne, Melinda B.; Mumford, Sunni L.; Segars, James H.; Armstrong, Alicia Y.

    2016-01-01

    Objective To compare cost and efficacy of tubal anastomosis to in vitro fertilization (IVF) in women who desired fertility after a tubal ligation. Design Cost-effectiveness analysis. Setting Not applicable. Patient(s) Not applicable. Intervention(s) Not applicable. Main Outcome Measure(s) Cost per ongoing pregnancy. Result(s) Cost per ongoing pregnancy for women after tubal anastomosis ranged from $16,446 to $223,482 (2014 USD), whereas IVF ranged from $32,902 to $111,679 (2014 USD). Across maternal age groups <35 and 35–40, years tubal anastomosis was more cost effective than IVF for ongoing pregnancy. Sensitivity analyses validated these findings across a wide range of ongoing pregnancy probabilities as well as costs per procedure. Conclusion(s) Tubal anastomosis was the most cost-effective approach for most women less than 41 years of age, whereas IVF was the most cost-effective approach for women aged ≥41 years who desired fertility after tubal ligation. A model was created that can be modified based on cost and success rates in individual clinics for improved patient counseling. PMID:26006734

  9. ESR1 rs9340799 Is Associated with Endometriosis-Related Infertility and In Vitro Fertilization Failure

    PubMed Central

    Paskulin, Diego Davila; Cunha-Filho, João Sabino; Paskulin, Livia Davila; Souza, Carlos Augusto Bastos; Ashton-Prolla, Patricia

    2013-01-01

    Estrogen receptor alpha has a central role in human fertility by regulating estrogen action in all human reproductive tissues. Leukemia inhibitory factor (LIF) expression, a cytokine critical for blastocyst implantation, is mediated by estrogen signaling, so we hypothesized that ESR1 gene polymorphisms might be candidate risk markers for endometriosis-related infertility and in vitro fertilization (IVF) failure. We included 98 infertile women with endometriosis, 115 infertile women with at least one IVF failure and also 134 fertile women as controls. TaqMan SNP assays were used for genotyping LIF (rs929271), MDM2 (rs2279744), MDM4 (rs1563828), USP7 (rs1529916), and ESR1 (rs9340799 and rs2234693) polymorphisms. The SNP ESR1 rs9340799 was associated with endometriosis-related infertility (P < 0.001) and also with IVF failure (P = 0.018). After controlling for age, infertile women with ESR1 rs9340799 GG genotype presented 4-fold increased risk of endometriosis (OR 4.67, 95% CI 1.84–11.83, P = 0.001) and 3-fold increased risk of IVF failure (OR 3.33, 95% CI 1.38–8.03, P = 0.007). Our results demonstrate an association between ESR1 rs9340799 polymorphism and infertile women with endometriosis and also with women who were submitted to IVF procedures and had no blastocyst implantation. PMID:24427778

  10. Evaluation of frozen thawed cauda epididymal sperms and in vitro fertilizing potential of bovine sperm collected from the cauda epididymal

    PubMed Central

    Chaveiro, A; Cerqueira, C; Silva, J; Franco, J; Moreira da Silva, F

    2015-01-01

    In the present study, the fertilizing potential of semen recovered from slaughtered bulls epididymis was evaluated after cryopreservation, by conventional techniques and flow cytometry methods. The cauda epididymal was dissected and sperm were recovered and evaluated for volume, sperm concentration, and membrane and acrosome integrity using a flow cytometer. Sperm fertility potential was tested by in vitro fertilization (IVF). For each bull, three trials of IVF were performed. Before freezing, on average, the sperm concentration was 216 ± 27.5 × 106 sperm/ml. Sperm viability averaged 86.5 ± 4%. The mean percentage of sperm with intact plasma membrane and acrosome before and after cryopreservation was 90.7 ± 2.9% and 90.8 ± 1.9% (P≥0.05), respectively. The fertilization rate using frozen/thawed epididymal semen averaged 64.1 ± 3.9% fertilization with no significant differences between bulls (P>0.05). For the bull considered as control, the fertilization rate was 72.2 ± 4.5%, differing significantly (P>0.05) from the frozen/thawed epididymal semen’s fertilization rate. In conclusion, it is possible to use in vitro techniques with cryopreserved spermatozoa obtained from bull’s epididymis using a controlled rate freezing method with a predetermined freezing curve, and with assessment of sperm’s viability by conventional techniques and flow cytometry methods, together with the fertilizing ability of cryopreserved epididymal spermatozoa. PMID:27175174

  11. Evaluation of Chang's culture medium for mouse in vitro fertilization and embryonic development.

    PubMed

    Ariff, B; Ng, S C; Mok, H; Lim, M N; Wong, P C; Shan, R

    1988-04-01

    Chang's medium [with and without human serum (HS)] was compared with T6 medium [with and without bovine serum albumin (BSA)] for in vitro fertilization (IVF) and development of two-cell mouse embryos to the blastocyst stage. Chang's medium without any supplementation gave significantly better fertilization rates (83.3%) than Chang's with 10% HS (76.4%) or T6 and BSA (76.6%) (P less than 0.01). In a separate experiment 87.7% of the two-cell mouse embryos developed to the blastocyst stage in Chang's medium, compared to 90.6% for T6 with BSA and 93.6% without BSA, respectively (P greater than 0.01). In Chang's medium supplemented with 10% HS, 76.6% of the embryos developed to the blastocyst stage and 17.2% stopped development after the morula stage. After 72 hr in vitro hatched trophoblast and inner-cell-mass cells from 26.5 and 30.8% of the embryos grown in Chang's medium (with and without HS) attached to the plastic culture dishes and grew to form a mixed monolayer of epithelioid and fibroblastic cells. Chang's medium can thus be successfully used for IVF and growth of mammalian embryos. Further, inner cell mass and trophoblast cell lines could be established for various reproductive studies using this medium.

  12. [Family planning in women with schizophrenia - case report on a schizophrenic patient with multiple pregnancy resulting from in vitro fertilization].

    PubMed

    Kapfhammer, Hans-Peter; Mayer, Christian

    2011-01-01

    Case report on a 31-year old woman with chronic schizophrenia turning to a gynaecological university outpatient clinic with her husband and asking for an in vitro fertilization. After referral to the psychiatric consultation service the patient's psychiatric and psychosocial history was taken. The issue of indication was settled by considering medicoethical aspects, general risks in respect of the course of illness in pregnancy, postpartum period, and parenthood, and the special psychological burden due to in vitro fertilization with the issues of multiple pregnancy and the dilemma of fetal reduction. The patient's course during a triple pregnancy, postpartum period and early parenthood is described. PMID:21672509

  13. Xenogenous fertilization of equine oocytes following recovery from slaughterhouse ovaries and in vitro maturation.

    PubMed

    Wirtu, G; Bailey, T L; Chauhan, M S; Parker, N A; Dascanio, J J; Gwazdauskas, F C; Ley, W B

    2004-01-15

    The in vitro production (IVP) of equine embryos using currently available protocols has met limited success; therefore investigations into alternative approaches to IVP are justified. The objective of this study was to evaluate the feasibility of xenogenous fertilization and early embryo development of in vitro matured (IVM) equine oocytes. Follicular aspirations followed by slicing of ovarian tissue were performed on 202 equine ovaries obtained from an abattoir. A total of 667 oocytes (3.3 per ovary) were recovered from 1023 follicles (recovery rate, 65%). Oocytes underwent IVM for 41 +/- 2 h (mean +/- S.D.), before being subjected to xenogenous gamete intrafallopian transfer (XGIFT). An average of 13 +/- 0.8 oocytes and 40x10(3) spermatozoa per oocyte were transferred into 20 oviducts of ewes. Fourteen percent of transferred oocytes (36/259) were recovered between 2 and 7 days post-XGIFT and 36% of those recovered displayed embryonic development ranging from the 2-cell to the blastocyst stage. Fertilization following XGIFT was also demonstrated by the detection of zinc finger protein Y (ZFY) loci. Ligation of the uterotubal junction (UTJ), ovarian structures, or the duration of oviductal incubation did not significantly affect the frequency of embryonic development or recovery of oocytes/embryos after XGIFT. In conclusion, equine embryos can be produced in a smaller non-equine species that is easier for handling. PMID:14662137

  14. Transvaginal oocyte retrieval controlled by vaginal probe for in vitro fertilization: a comparative study.

    PubMed

    Feldberg, D; Goldman, J A; Ashkenazi, J; Shelef, M; Dicker, D; Samuel, N

    1988-06-01

    Of 142 patients who initiated in vitro fertilization cycles, 52 underwent oocyte collection by laparoscopy, 58 by the ultrasonically guided transvesical route, both under general or local anesthesia and 36 by the ultrasonically transvaginal route under local anesthesia. When the three methods were compared the results did not differ significantly. An average of 6.4, 6.2, and 5.7 oocytes, respectively, were retrieved. Cleavage rates were 82.6, 79.4, and 81.6%, respectively, while 20.2, 22.6, and 21.1% clinical pregnancies per cycle were obtained. Complications with this procedure such as exacerbation of previous pelvic inflammatory disease, urinary tract infections, transient hematuria, and mild hemoperitoneum were rare. In general, patients greatly preferred the transvaginal method. In addition to providing greater comfort for the patient, this method requires no general anesthesia and is less expensive than other methods. Our initial experience demonstrates that in vitro fertilization can be performed successfully through this simplified alternative to oocyte retrieval.

  15. Potential Pathophysiological Mechanisms of the Beneficial Role of Endometrial Injury in In Vitro Fertilization Outcome.

    PubMed

    Siristatidis, Charalampos; Vrachnis, Nikos; Vogiatzi, Paraskevi; Chrelias, Charalampos; Retamar, Andrea Quinteiro; Bettocchi, Stefano; Glujovsky, Demián

    2014-03-01

    Successful embryo implantation is a complex process that involves multiple biological mechanisms and reciprocal interactions between the embryo and the proliferated endometrium. In this review, we provide an informative contribution on the pathways underlying the beneficial nature of endometrial injury toward improving implantation rates of embryos conceived and through in vitro fertilization. The evidence published to date are in favor of inducing local endometrial injury in the preceding cycle of ovarian stimulation to improve pregnancy outcomes in women with unexplained and recurrent implantation failure. Endometrial injury triggers a series of biological responses but the findings suggest that no particular pathway is solely adequate to explain the association between trauma and improved pregnancy rates rather than a cluster of events in response to trauma which benefits embryo implantation in ways both known and unknown to the scientific community. PMID:24604231

  16. The creation of "monsters": the discourse of opposition to in vitro fertilization in Poland.

    PubMed

    Radkowska-Walkowicz, Magdalena

    2012-12-01

    In Poland, there is a campaign to criminalise in vitro fertilization, led by the Catholic church. This article explores how this campaign makes "monsters" of IVF children in its discourse, that is, embodiments of "the other" in the sense of Frankenstein's monster. Basing the analysis primarily on Catholic mass media publications, the article investigates the discursive strategies employed to oppose IVF, most notably by the Catholic clergy and activists and journalists associated with the Church. They attribute "monstrosity" to the children in the following ways: physical (possible bodily deformity), psychological (survivor syndrome, identity crisis), social (loneliness, uncertain place in family relations), and ethical (a life burdened with the deaths of many embryos). Although the world of families with IVF does not provide examples of children who could be considered monsters in any of these terms, these arguments have become the primary reasons given for banning IVF.

  17. Results of in vitro fertilization in women with antisperm antibodies in serum, cervical mucus, and follicular fluid.

    PubMed

    Potashnik, G; Kleinman, D; Insler, V; Albotiano, S; Glezerman, M; Meizner, I

    1988-08-01

    This study was undertaken to investigate the presence of antisperm antibodies (ASA) in serum, cervical mucus, and follicular fluid (FF) of women undergoing in vitro fertilization and embryo transfer (IVF-ET). IgG and IgA ASA directed mostly against sperm head were found at similar concentrations in serum, cervical mucus, and FF of 2 of 34 patients. Ninety-one percent fertilization and 100% cleavage rates, respectively, were observed in one of the two patients. No fertilization occurred in the second patient. In both women, in vitro sperm penetration tests revealed hostile mucus and repeated postcoital tests were poor. It is concluded that the sperm-cervical mucus penetration test and mucus ASA measurements are useful in establishing the diagnosis of immunological infertility.

  18. Localization of fucosyl glycoconjugates in human oocytes following insemination for in vitro fertilization.

    PubMed

    Tam, P P; Loong, E P; Chiu, T T

    1990-06-01

    Human oocytes that failed to cleave after insemination were examined for the presence of fucosyl glycoconjugates in the perivitelline space by staining with Ulex europeaus lectin conjugated to fluorescein isothiocynate. Oocytes that formed two or three pronuclei following first insemination always exhibited positive lectin staining similar to that observed with in vitro fertilized mouse oocytes. Among those oocytes that failed to form any pronuclei after the first insemination attempt, only 5% contained lectin positive substances in the perivitelline space. Upon reinsemination, a higher percentage of those oocytes produced lectin-positive materials, although pronuclei were still absent. The appearance of fucosyl glycoconjugates in these oocytes might be the result of the release of cortical granules triggered by sperm penetration or, more likely, due to spontaneous granule discharge in senescent oocytes.

  19. Royal Jelly alleviates sperm toxicity and improves in vitro fertilization outcome in Stanozolol-treated mice

    PubMed Central

    Shalizar Jalali, Ali; Najafi, Gholamreza; Hosseinchi, Mohammadreza; Sedighnia, Ashkan

    2015-01-01

    Background: Stanozolol (ST) is a synthetic anabolic-androgenic steroid often abused by athletes. An increasing body of evidence points towards the role of ST misuses in the pathogenesis of a wide range of adverse effects including reprotoxicity. Objective: The aim of this study was to analyze the possible reproprotective effect of royal jelly (RJ) as an efficient antioxidant in ST-treated mice. Materials and Methods: Adult male mice were divided into four groups (n=5). Two groups of mice received ST (4.6 mg/kg/day) via gavage for 35 days. RJ was given orally to one of these groups at the dose level of 100 mg/kg body weight per day synchronously. Untreated control group and RJ-only treated group were also included. Epididymal sperm characteristics and in vitro fertilizing capacity were evaluated after 35 days. Results: ST treatment caused a significant (p<0.05) decrease in sperm count and motility and fertilization rate along with poor blastocyst formation and increased sperm DNA damage. Moreover, the incidence of apoptosis and abnormality in spermatozoa was significantly (p<0.05) higher in ST-exposed mice than those of control. The above-mentioned parameters were restored to near normal level by RJ co-administration. Conclusion: Data from the current study suggest that RJ has a potential repro-protective action against ST-induced reproductive toxicity in mice. However, clinical studies are warranted to investigate such an effect in human subjects. PMID:25653671

  20. Deep venous thrombosis in a patient undergoing In-vitro fertilization with oocyte donation

    PubMed Central

    Mahajan, Nalini; Naidu, Padmaja; Gupta, Shalu; Rani, Kumkum

    2015-01-01

    Deep venous thrombosis (DVT) has been reported extensively following ovarian hyperstimulation syndrome during in-vitro fertilization (IVF). Pregnancy per se increases the risk of DVT due to a hypercoagulable state. The long-term use of hormone replacement therapy (HRT) is another critical factor associated with DVT in women. However, an association between the short-term use of HRT in oocyte donation (OD) cycles and DVT has not yet been reported. We present a case of 43-year-old woman who developed DVT after IVF-OD. DVT was diagnosed at 7 weeks of pregnancy and was managed with low-molecular-weight heparin. We suggest that even a short-term use of HRT should be considered a risk factor for DVT especially in the presence of additional risk factors such as obesity. The patient had an uneventful recovery and delivered three healthy though preterm babies. PMID:26538864

  1. Brain Tumor in an In Vitro Fertilization-Facilitated Pregnancy: Fourth Ventricle Anaplastic Ependymoma in the Second Trimester.

    PubMed

    Ryskeldiyev, Nurzhan; Olenbay, Gabit; Auezova, Raushan; Killeen, Tim; Aldiyarova, Nurgul; Akhmetzhanova, Zauresh; Cesnulis, Evaldas; Akshulakov, Serik

    2016-06-01

    We present a case of fourth ventricle anaplastic ependymoma in a pregnancy which was the first result of three rounds of in vitro fertilization (IVF) and embryo transfer. Whether hormonal treatment can directly or indirectly precipitate brain tumors to develop or become symptomatic is unclear. PMID:27330925

  2. GnRH antagonist in in vitro fertilization: where we are now.

    PubMed

    Shapiro, D B; Mitchell-Leef, D

    2003-10-01

    This review focuses on the recent literature concerning the use of GnRH antagonists in ovulation induction for in vitro fertilization (IVF). The GnRH antagonists, ganirelix acetate (Orgalutran/Antagon) and cetrorelix (Cetrotide), have come into increasingly common use since their release in the last 3 years. This class of GnRH analogue has several potential advantages over GnRH agonists. Among these advantages are: 1) shorter duration of injectable drug treatment, 2) decreased gonadotropin requirement per cycle, 3) improved patient convenience and 4) lower overall treatment cost. As clinicians gain experience with these drugs, optimal treatment paradigms will likely emerge. This review will discuss current strategies and potential applications for the GnRH antagonists.

  3. Effectiveness of nursing counseling on coping and depression in women undergoing in vitro fertilization.

    PubMed

    Gürhan, Nermin; Oflaz, Fahriye; Atici, Derya; Akyüz, Aygol; Vural, Gulsen

    2007-04-01

    The purpose of present study was to evaluate the effectiveness of counseling provided by nurses on depression and coping strategies of infertile women undergoing in vitro fertilization (N=67). Of the 84 women who were interviewed, 30 were accepted as a comparison group, and 37 were included in the study group. The study group women were given counseling in addition to routine nursing care services, including group education and individual interviews about treatment and coping strategies. The nurses also provided support by accompanying the women during the invasive procedures. The Beck Depression Inventory and Jalowiec's Coping Strategies Form were used for measurements. All the women were using emotional coping and had moderate depression prior to the study. There was no statistically significant difference between the comparison and study groups before or after the counseling with respect to depression and coping strategies. Parameters to evaluate the efficacy of counseling are discussed.

  4. Spectral interferometry for morphological imaging in in vitro fertilization (IVF) (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Zhu, Yizheng; Li, Chengshuai

    2016-03-01

    Morphological assessment of spermatozoa is of critical importance for in vitro fertilization (IVF), especially intracytoplasmic sperm injection (ICSI)-based IVF. In ICSI, a single sperm cell is selected and injected into an egg to achieve fertilization. The quality of the sperm cell is found to be highly correlated to IVF success. Sperm morphology, such as shape, head birefringence and motility, among others, are typically evaluated under a microscope. Current observation relies on conventional techniques such as differential interference contrast microscopy and polarized light microscopy. Their qualitative nature, however, limits the ability to provide accurate quantitative analysis. Here, we demonstrate quantitative morphological measurement of sperm cells using two types of spectral interferometric techniques, namely spectral modulation interferometry and spectral multiplexing interferometry. Both are based on spectral-domain low coherence interferometry, which is known for its exquisite phase determination ability. While spectral modulation interferometry encodes sample phase in a single spectrum, spectral multiplexing interferometry does so for sample birefringence. Therefore they are capable of highly sensitive phase and birefringence imaging. These features suit well in the imaging of live sperm cells, which are small, dynamic objects with only low to moderate levels of phase and birefringence contrast. We will introduce the operation of both techniques and demonstrate their application to measuring the phase and birefringence morphology of sperm cells.

  5. Impact of ketorolac administration around ovarian stimulation on in vivo and in vitro fertilization and subsequent embryo development.

    PubMed

    Jee, Byung Chul; Youm, Hye Won; Lee, Jae Ho; Kim, Jee Hyun; Suh, Chang Suk; Kim, Seok Hyun

    2013-05-01

    We performed this study to investigate the effect of ketorolac (a non-steroidal anti-inflammatory drug) administration around ovarian stimulation on in vivo and in vitro fertilization process. Sixty-four female mice (ICR) were injected with ketorolac (0, 7.5, 15 and 30 µg/d) for 3 d starting from the day of eCG treatment. In experiment 1, 41 mice were triggered by hCG and then mated; two-cell embryos were obtained and in vitro development up to blastocyst was observed. In experiment 2, 23 mice were triggered by hCG and mature oocytes were collected; in vitro fertilization rate and subsequent embryo development up to blastocyst was recorded. In experiment 1, the blastocyst-forming rates per in vivo fertilized two-cell embryo showed an inverse relationship with a dosage of ketorolac (97.6%, 64.2%, 35.4% and 25.9%). In experiment 2, degenerated oocytes were frequently observed in a dose-dependent manner (4.3%, 22.9%, 22.4% and 75.0%). Lower fertilization rates were noted in all the three ketorolac-treating groups; blastocyst-forming rate was significantly lower in 30-µg-treating group when compared with the control group. Administration of ketorolac around ovarian stimulation significantly affects the development of in vivo fertilized embryo in a dose-dependent manner. High-dose ketorolac could result in a poor oocyte quality and decreased embryo developmental competence.

  6. Results of planned in-vitro fertilization programming through the pre-administration of the oestrogen-progesterone combined pill.

    PubMed

    Cohen, J; Debache, C; Solal, P; Serkine, A M; Achard, B; Boujenah, A; Pez, J P; Paris, X; Robert, J; Loffredo, V

    1987-01-01

    The use of an oestrogen-progesterone combined pill permits the induction of ovulation in the absence of any developing follicle. Two treatments were compared. In the first, patients received no prior treatment before stimulation. In the second, combined oestrogen-progesterone treatment was given during approximately two menstrual cycles prior to stimulation. No differences between the two groups were found in relation to oocyte maturity, fertilization in vitro, cleavage, replacement and pregnancy. Fewer luteinizing hormone surges occurred in patients pre-treated with steroids. The utilization of the oestrogen-progesterone combined pill prior to induction of ovulation facilitates the forward planning of patients for in-vitro fertilization. PMID:3106405

  7. Apparatus for the in vitro fertilization and culture of human oocytes.

    PubMed

    Testart, J; Lassalle, B; Frydman, R

    1982-09-01

    An apparatus for in vitro fertilization and culture of mammalian eggs that permits close control over variations in temperature, lighting, and pH has been developed to imitate natural conditions as far as possible. All handling of the oocyte and zygote are performed inside a baby incubator which has a system of air saturation using water vapor. A binocular dissecting microscope and an inverted compound microscope are built into the enclosure to permit simultaneous observation of the egg at widely different magnifications. Lighting is restricted to the longer wavelengths, which do not harm mammalian eggs. The microscope is equipped with a camera system. A gas mixture of 5% CO2, 5% O2, and 90% N2 is delivered at a pressure of 1 bar by a double reduction valve arrangement. The gas passes into the incubator through a filter and is humidified and warmed inside the incubator by bubbling through a bottle of sterile distilled water. The gas is finally distributed by a series of culture tubes. The culture tubes are usually kept in darkness; their position is changed as the development of the egg progresses to take advantage of the slight temperature differentials within the apparatus. Less than 1 minute after puncture of the follicle, the fluid is transferred to the incubator and is examined in a petri dish using the stereomicroscope. The oocyte is immediately transferred to a culture tube containing medium and placed at 36.8 degrees Celsius. 16-20 hours after insemination the egg is transferred under the stereomicroscope by pipetting directly into a new tube, where the medium is supplemented by a small quantity of maternal serum. It is kept at 37.2 degrees Celsius. After 2-4 days of culture in vitro, if normal development is observed, the egg is transferred to the patient's uterus. The variation in temperature at any given point in the apparatus is less than .2 degrees Celsius. 114 human oocytes have been collected, of which 58% were fertilized. 82% of the fertilized eggs

  8. Pertinence of apoptosis markers for the improvement of in vitro fertilization (IVF).

    PubMed

    Haouzi, D; Hamamah, S

    2009-01-01

    In assisted reproductive technology (ART), the pregnancy and birth rates following in vitro fertilization (IVF) attempts are still low. Recently, apoptotic markers have been suggested as new criteria for oocyte and embryo quality selection. Many studies have provided evidence that poor oocyte and embryo quality can be associated with apoptosis. The aim of this review is to summarize our current knowledge on the apoptotic process in oocytes and embryos, and focus on the possibility for using apoptotic markers as a reliable and predictive marker to select competent oocytes and embryos during IVF. Moreover, it is currently accepted that IVF failures, linked to poor embryo quality, are, in part, associated with suboptimal in vitro culture conditions. Here, we also review the current state of knowledge concerning how the genetic control of apoptosis during folliculogenesis and pre-implantation embryonic development is affected by in vitro culture conditions during IVF. In the future, identification of apoptotic markers in ART for oocyte and embryo selection should result in the development of new agonistic or antagonistic molecules of apoptosis by medicinal chemistry.

  9. High-dose follicle-stimulating hormone (FSH) ovarian stimulation in low-responder patients for in vitro fertilization.

    PubMed

    Hofmann, G E; Toner, J P; Muasher, S J; Jones, G S

    1989-10-01

    Follicle-stimulating hormone (FSH) was used in high doses (6 ampoules/day:6FSH) for ovarian hyperstimulation for in vitro fertilization in women with a previous poor response to stimulation with the equivalent of "4FSH." Luteinizing hormone levels did not differ between stimulations, but both FSH and estradiol levels were higher in the 6FSH compared to the 4FSH cycle. There were fewer cancellations in the 6FSH cycle, but similar numbers of preovulatory oocytes were retrieved, fertilized, and transferred. The pregnancy rates per attempt and retrieval were higher in the 6FSH cycle. We conclude that raising and maintaining FSH levels during stimulation in low responders reduced cancellations and may improve in vitro fertilization outcome.

  10. The use of transvaginal ultrasound to aspirate bilateral hydrosalpinges prior to in vitro fertilization: a case report.

    PubMed

    Russell, J B; Rodriguez, Z; Komins, J I

    1991-08-01

    Transvaginal ultrasound has recently been introduced diagnostically for follicular monitoring, early pregnancy localization (1), pelvic structure identification, early fetal development (2), detection of ovarian enlargement in postmenopausal women (3), and detection of endometrial carcinoma (4). Transvaginal ultrasound has had a major impact on patient monitoring and treatment for in vitro fertilization and embryo transfer (IVF/ET). Therapeutically transvaginal ultrasound has been introduced for oocyte retrieval and selective pregnancy reduction. The purpose of this paper is to present the therapeutic use of transvaginal ultrasound to aspirate large bilateral hydrosalpinges which complicated embryo transfer during an in vitro fertilization cycle.

  11. Through the smoke: Use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility

    SciTech Connect

    Camlin, Nicole J.; McLaughlin, Eileen A.; Holt, Janet E.

    2014-12-15

    A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents’ in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes. - Highlights: • In vivo exposure to individual cigarette smoke chemicals alters female fertility. • The use of in vitro models in determining molecular mechanisms • Whole cigarette smoke inhalation animal models negatively affect ovarian function.

  12. INHIBITION OF IN VITRO FERTILIZATION IN THE HAMSTER BY ANTIBODIES RAISED AGAINST THE RAT SPERM PROTEIN SP22

    EPA Science Inventory

    INHIBITION OF IN VITRO FERTILIZATION IN THE HAMSTER BY ANTIBODIES RAISED AGAINST THE RAT SPERM PROTEIN SP22. SC Jeffay*, SD Perreault, KL Bobseine*, JE Welch*, GR Klinefelter, US EPA, Research Triangle Park, NC.
    SP22, a rat sperm membrane protein that is highly-correlated w...

  13. A morphologic study of unfertilized oocytes and abnormal embryos in human in vitro fertilization.

    PubMed

    Bałakier, H; Casper, R F

    1991-04-01

    The morphology of human, unfertilized oocytes and abnormal embryos cultured in vitro for 48-72 hr was examined in an attempt to learn more about oocyte maturation and reproductive failure in in vitro fertilization (IVF). About 21% of the unfertilized oocytes were totally degenerated. The majority (56%) of the remaining oocytes was arrested at the metaphase II stage. They contained coherent chromosomal plates and had extruded the first polar body with nuclear material. About 13% of oocytes underwent spontaneous activation. In most of these cases the second polar body was retained and many subnuclei or one big nucleus was formed. Five percent of metaphase II oocytes penetrated by sperm were not activated, likely as a result of oocyte immaturity. The developmental ability of abnormal embryos was poor. Several one-cell-stage zygotes were arrested at the pronuclear stage or at mitosis of the first mitotic division. Polyspermic embryos, especially those which contained four or more pronuclei, did not divide or formed uneven, multinucleated blastomeres. However, some triploid and tetraploid embryos often appeared normal morphologically despite their lethal chromosomal abnormalities.

  14. Monochorionic triamniotic triplets following conventional in vitro fertilization and blastocyst transfer

    PubMed Central

    Gurunath, Sumana; Makam, Adinarayana; Vinekar, Sriprada; Biliangady, Reeta H.

    2015-01-01

    Multiple pregnancy in in vitro fertilization (IVF) is on the decline with a reduction in number of embryos transferred. But the risk of monozygotic splitting persists. The risk of monozygotic twinning in women undergoing IVF is reported to be twice that of natural conception, and monochorionic triplets are even rarer at 100 times more than natural conception. We report a case of monochorionic triamniotic (MCTA) triplets following conventional IVF and blastocyst transfer without zona manipulation. This report highlights the possibility of zygotic splitting in IVF in young couples with no family history, in centers with good experience with blastocyst transfer. MCTA triplets carry a high risk of perinatal mortality and morbidity and need multidisciplinary care. Prevention and prediction of zygotic splitting ought to be realized with better reporting and identification of possible risk factors. PMID:25838751

  15. Effect of antral follicle count (AFC) in beef heifers on in vitro fertilization/production (IVF/IVP)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our objective has been to compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the 4th year of the study with years 1 to 3 reported individually. For this report, we add data for the 4th year and present a combined an...

  16. International regulatory landscape and integration of corrective genome editing into in vitro fertilization.

    PubMed

    Araki, Motoko; Ishii, Tetsuya

    2014-11-24

    Genome editing technology, including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas, has enabled far more efficient genetic engineering even in non-human primates. This biotechnology is more likely to develop into medicine for preventing a genetic disease if corrective genome editing is integrated into assisted reproductive technology, represented by in vitro fertilization. Although rapid advances in genome editing are expected to make germline gene correction feasible in a clinical setting, there are many issues that still need to be addressed before this could occur. We herein examine current status of genome editing in mammalian embryonic stem cells and zygotes and discuss potential issues in the international regulatory landscape regarding human germline gene modification. Moreover, we address some ethical and social issues that would be raised when each country considers whether genome editing-mediated germline gene correction for preventive medicine should be permitted.

  17. Community attitudes to artificial insemination by husband or donor, in vitro fertilization, and adoption.

    PubMed

    Rowland, R; Ruffin, C

    1983-09-01

    Although reproductive medicine is rapidly advancing within society, few studies have been conducted in this area on the opinions and attitudes of people in the community. A survey of the attitudes of 104 Geelong residents, selected by a random sampling technique, was conducted, in order to assess community attitudes to adoption, artificial insemination using husband and donor sperm, and in vitro fertilization. The results indicate that attitudes in general are positive, supporting the pro-natalist bias of society. AID was seen as the most problematic technique used, and was given less support in general than the other techniques. A surprising number of respondents felt that single and homosexual groups should have access to these techniques. Further results are discussed, including those bearing on the possibility of establishing a National Register for AID donor-offspring information.

  18. Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

    PubMed

    Pereira, A F; Melo, L M; Freitas, V J F; Salamone, D F

    2015-08-01

    In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of γH2AX foci or area were detected among the treatments. γH2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development.

  19. Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

    PubMed

    Pereira, A F; Melo, L M; Freitas, V J F; Salamone, D F

    2015-08-01

    In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of γH2AX foci or area were detected among the treatments. γH2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development. PMID:24735637

  20. Bovine in vitro reproduction models can contribute to the development of (female) fertility preservation strategies.

    PubMed

    Langbeen, An; De Porte, Hannelore F M; Bartholomeus, Esther; Leroy, Jo L M R; Bols, Peter E J

    2015-09-01

    Recent increases in the number of successful cancer treatments have stimulated interest in fertility preservation strategies in women of reproductive age and in prepubertal girls. However, research on the application of such programs under clinical conditions suffers from the scarce availability of human tissue for research purposes and from concurrent relevant ethical issues. To partly address this problem, this review focuses on the possibilities of ruminant in vitro models providing additional insights into several aspects of fertility preservation, ranging from preantral follicle collection to oocyte and follicle cryopreservation, to noninvasive quality assessment, and to follicle culture. After a brief introduction, we discuss currently available techniques involved in (human) fertility preservation, together with their inherent advantages and limitations. On the basis of literature, we describe specific points for improvement or urgent additional research, such as (1) the lack of noninvasive methods to assess viability and developmental capacity of preantral follicles (either isolated or "in situ"); (2) autotransplantation and cryopreservation of ovarian cortex and follicles; (3) ischemia, follicular burnout, and graft rejection as major causes of preantral follicle loss; and (4) the development of routine in vitro follicle culture methods. Within each section, an overview is given of similar available techniques in (ruminant) assisted reproduction, with suggestions as to where and how these research models might contribute to fill the identified gaps. After the identification of the remaining issues in the development of integrated fertility preservation strategies, available ruminant in vitro models are introduced, described, and matched to these challenges to define common grounds for reproductive research. Ruminant in vitro models are increasingly considered as being very relevant for human preimplantation reproductive research. Because ruminant in vitro

  1. Transvaginal sonographically controlled ovarian puncture for oocyte retrieval for in vitro fertilization.

    PubMed

    Cohen, J; Debache, C; Pez, J P; Junca, A M; Cohen-Bacrie, P

    1986-10-01

    Two hundred twenty-two patients took part in a trial of follicle puncture via the transvaginal route under sonographic control for the purpose of in vitro fertilization (IVF). Induction protocols were mainly human menopausal gonadotropin (hMG) + human chorionic gonadotropin (hCG) and clomiphene + hMC + hCG. In 79.7% oocyte aspiration could be achieved without difficulty via the transvaginal route. An average number of 4.7 oocytes per attempt was obtained; 10.7% evolutive pregnancies were obtained. No major incident was noted. This technique offers several crucial advantages: it reduces surgical risk, reduces the length of the patient's stay in hospital as well as the overall cost of the procedure, and it also makes possible puncture in some cases hitherto regarded as excluded. PMID:3537171

  2. [Discussion on the impact of acupuncture for the pregnancy of in vitro fertilization-embryo transfer].

    PubMed

    Xie, Zhengyun; Yao, Bing; Chen, Li; Mu, Yanyun; Cheng, Jie; Li, Qian; Gao, Zhao; Xia, You-bing

    2016-05-01

    The impact of acupuncture for the pregnancy of in vitro fertilization-embryo transfer (IVF-ET) is discussed in the paper. Nowadays there are various conclusions about the impact of acupuncture for IVF-ET, and it may result from the differences in research designs. The effect is closely related to the demographic and clinical characteristics of subjects, such as age, the diagnosis of barrenness, blood flow index of uterine spiral arteries, the cycle of IVF, etc. Besides, the efficacy is influenced by treatment based on syndrome differentiation or not, the frequency and course of acupuncture in both the treating group and the control group, etc. If more reasonable design is achieved in the further study based on them, more reliable evidence will be provided for the effect and mechanism of the pregnancy of IVF-EF by acupuncture. PMID:27509598

  3. Clinical and Structural Features of Sperm Head Vacuoles in Men Included in the In Vitro Fertilization Programme

    PubMed Central

    Štrus, Jasna; Tušek Žnidarič, Magda; Knez, Katja; Vrtacnik Bokal, Eda; Virant-Klun, Irma

    2014-01-01

    The human sperm head vacuoles and their role in male infertility are still poorly understood. The aim of this study was to identify the clinical and ultrastructural features of human sperm head vacuoles in men included in the in vitro fertilization programme: men with normal (normozoospermia) and impaired sperm morphology (teratozoospermia). The sperm samples were observed under 6000-time magnification using motile sperm organelle morphology examination (MSOME). The proportion of sperm with head vacuoles was evaluated and related to the outcome of in vitro fertilization. The sperm of men with impaired sperm morphology was characterized by a higher proportion of sperm head vacuoles. The sperm head vacuoles were related to impaired semen quality (sperm concentration, motility, and morphology) but were not influenced by male factors (semen volume, height, age, weight, or body mass index). Moreover, sperm head vacuoles were related to impaired fertilization rate merely after classical in vitro fertilization (IVF), while there was no relation to pregnancy. In a subgroup of men, the sperm was fixed and observed by transmission electron microscopy (TEM). The ultrastructural study revealed that sperm head vacuoles are large nuclear indentations of various sizes and positions, packed with membranous material organized in membrane whorls (MW). PMID:24818161

  4. Effect of Paternal Age on Reproductive Outcomes of In Vitro Fertilization.

    PubMed

    Wu, Yixuan; Kang, Xiangjin; Zheng, Haiyan; Liu, Haiying; Liu, Jianqiao

    2015-01-01

    Although the adverse effects of maternal aging on reproductive outcomes have been investigated widely, there is no consensus on the impact of paternal age. Therefore, we investigated the effect of paternal age on reproductive outcomes in a retrospective analysis of 9,991 in vitro fertilization (IVF) cycles performed at the Reproductive Medicine Center of the Third Affiliated Hospital of Guangzhou Medical University (China) between January 2007 and October 2013. Samples were grouped according to maternal age [<30 (3,327 cycles), 30-34 (4,587 cycles), and 35-38 (2,077 cycles)] and then subgrouped according to paternal age (<30, 30-32, 33-35, 36-38, 39-41, and ≥42). The groups did not differ in terms of fertilization rate, numbers of viable and high-quality embryos and miscarriage rate when controlling maternal age (P >0.05). Chi-squared analysis revealed that there were no differences in implantation and pregnancy rates among the different paternal age groups when maternal age was <30 and 35-38 years (P >0.05). However, implantation and pregnancy rates decreased with paternal age in the 31-34 y maternal age group (P <0.05). Our study indicates that paternal age has no impact on fertilization rate, embryo quality at the cleavage stage and miscarriage rate. For the 30-34 y maternal age group, the implantation rate decreased with increased paternal age, with the pregnancy rate in this group being significantly higher in the paternal <30 y and 30-32 y age groups, compared with those in the 36-38 y and 39-41 y groups.

  5. Effect of Paternal Age on Reproductive Outcomes of In Vitro Fertilization

    PubMed Central

    Zheng, Haiyan; Liu, Haiying; Liu, Jianqiao

    2015-01-01

    Although the adverse effects of maternal aging on reproductive outcomes have been investigated widely, there is no consensus on the impact of paternal age. Therefore, we investigated the effect of paternal age on reproductive outcomes in a retrospective analysis of 9,991 in vitro fertilization (IVF) cycles performed at the Reproductive Medicine Center of the Third Affiliated Hospital of Guangzhou Medical University (China) between January 2007 and October 2013. Samples were grouped according to maternal age [<30 (3,327 cycles), 30–34 (4,587 cycles), and 35–38 (2,077 cycles)] and then subgrouped according to paternal age (<30, 30–32, 33–35, 36–38, 39–41, and ≥42). The groups did not differ in terms of fertilization rate, numbers of viable and high-quality embryos and miscarriage rate when controlling maternal age (P >0.05). Chi-squared analysis revealed that there were no differences in implantation and pregnancy rates among the different paternal age groups when maternal age was <30 and 35–38 years (P >0.05). However, implantation and pregnancy rates decreased with paternal age in the 31–34 y maternal age group (P <0.05). Our study indicates that paternal age has no impact on fertilization rate, embryo quality at the cleavage stage and miscarriage rate. For the 30–34 y maternal age group, the implantation rate decreased with increased paternal age, with the pregnancy rate in this group being significantly higher in the paternal <30 y and 30–32 y age groups, compared with those in the 36–38 y and 39–41 y groups. PMID:26352861

  6. Cleavage capability of water buffalo follicular oocytes classified by cumulus cells and fertilized in vitro.

    PubMed

    Suzuki, T; Singla, S K; Sujata, J; Madan, M L

    1991-06-01

    Water buffalo (Murrah) oocytes were collected from ovaries obtained from the slaughter house. They were classified according to the character of the cumulus cells under a stereomicroscope, and cultured in 25 mM Hepes buffered Tissue Culture Medium-199 (TCM-199) supplemented with 5% estrous water buffalo serum in an atmosphere containing 5% CO2 in air at 39 degrees C. After 20-24 hr of in vitro maturation, the oocytes were fertilized using capacitated sperm obtained from 4 different bulls. For cleavage the oocytes were cultured at 39 degrees C in TCM-199 supplemented with 1% estrous water buffalo serum and in an atmosphere containing 5% CO2 in air. The good oocytes, with compact and dense cumulus cells cleaved significantly higher (p less than 0.01, 67.3%), than those of fair. partially naked oocytes with thin cumulus layers (27.5%, 25/91) or small remnants of cumulus cells and poor naked oocytes (3/100). A substantial variation cumulus layers (27.5% 25/91) or small remnants of cumulus cells and poor naked oocytes (3/100). A substantial variation in fertilization and developmental rates (16.0% to 43.8%) was observed among 4 different bulls. Late non-surgically into 14 buffalo recipients on day 6 or 7 of their estrous cycle. One recipient was diagnosed to be pregnant by rectal palpation on day 60 and confirmed to be so on day 90 post-estrus.

  7. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

    PubMed Central

    Bouillon, Céline; Léandri, Roger; Desch, Laurent; Ernst, Alexandra; Bruno, Céline; Cerf, Charline; Chiron, Alexandra; Souchay, Céline; Burguet, Antoine; Jimenez, Clément; Sagot, Paul; Fauque, Patricia

    2016-01-01

    In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium—SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge – 270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05). The culture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development. PMID:27008092

  8. Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

    PubMed

    Bouillon, Céline; Léandri, Roger; Desch, Laurent; Ernst, Alexandra; Bruno, Céline; Cerf, Charline; Chiron, Alexandra; Souchay, Céline; Burguet, Antoine; Jimenez, Clément; Sagot, Paul; Fauque, Patricia

    2016-01-01

    In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium--SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05). The culture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

  9. Clinical Outcomes of In Vitro Fertilization among Chinese Infertile Couples Treated for Syphilis Infection.

    PubMed

    Wang, Jing; Zhao, Xiaomiao; Yuan, Ping; Fang, Tingfeng; Ouyang, Nengyong; Li, Ruiqi; Ou, Songbang; Wang, Wenjun

    2015-01-01

    To compare the clinical outcomes of infertile patients with and without syphilis after in vitro fertilization and embryo transfer (IVF-ET), in this case-control study, 320 infertile couples were enrolled and divided into syphilis (n = 160) and control groups (n = 160). The primary IVF outcomes were the clinical pregnancy rate and the birth of a healthy baby. All syphilis patients received the standard anti-syphilis treatment before undergoing IVF/ICSI. Our results showed that the endometrial thickness of the syphilis group was greater than that of the control group following hCG injection (16.9±5.4 vs. 13.0±4.7 mm, P<0.001). The numbers of normally fertilized eggs and normally cleaved fertilized eggs and the implantation rate were 6.8±4.8, 6.3±4.7 and 24.2%, respectively, for the syphilis group and 8.3±4.6, 8.1±4.6 and 34.4%, respectively, for the control group, and these values were significantly different between the groups. The clinical pregnancy rate was lower in the syphilis group compared with that in the control group (43.8% vs. 55.6%, P = 0.03). Lower offspring birth weight was observed in the infected male group compared with those in the infected female (2.7±0.4 vs. 3.0±0.4 kg, P = 0.01) and infected couple groups (2.7±0.4 vs. 3.1±0.5 kg, P = 0.007). Therefore, syphilis infection reduces the clinical pregnancy rate after IVF/ICSI.

  10. Pericentric inversion of chromosome 9 causing infertility and subsequent successful in vitro fertilization

    PubMed Central

    Muthuvel, Arun; Ravindran, Manipriya; Chander, Aravind; Subbian, Chandralekha

    2016-01-01

    One of the most common and benign variants of normal human karyotype is pericentric inversion of chromosome 9 (inv[9][p11q13]). Despite being categorized as a normal variant, there are several reports of its association with various disease conditions. Here, we report a 27 year old female, who presented to us with primary infertility. The woman was diagnosed with inv (9)(p11q13) which was acknowledged as the reason for her otherwise unexplained infertility. The couple thereupon underwent in vitro fertilization using donor oocyte resulting in live birth. The clinical significance of this minor chromosomal rearrangement, need for genetic counseling, and subsequent reproductive guidance is highlighted in this report. PMID:27226692

  11. Short-term use of gonadotropin-releasing hormone agonist (leuprolide) for in vitro fertilization.

    PubMed

    Katayama, K P; Roesler, M; Gunnarson, C; Stehlik, E; Jagusch, S

    1988-12-01

    A common problem encountered by in vitro fertilization (IVF) programs is the premature occurrence of the spontaneous luteinizing hormone (LH) surge during ovarian stimulation cycles. Administration of gonadotropin-releasing hormone agonists (GnRH-a) for 2 to 3 weeks produces a state of hypogonadotropic hypogonadism, thus allowing ovarian stimulation to proceed uncomplicated by a spontaneous LH surge. We have elected to treat seven patients with GnRH-a in a "short-term" protocol, with GnRH-a initiated on cycle day 3 along with exogenous gonadotropins. In this series, we found that the spontaneous LH surge was abolished, while ovarian responsiveness seemed to be improved. These results suggest that the initial surge of gonadotropins elicited by GnRH-a administration may enhance ovarian stimulation and that spontaneous LH surge is blocked when GnRH-a and exogenous gonadotropins are initiated concomitantly.

  12. [A Case of Paradoxical Cerebral Embolism Developed during in vitro Fertilization Treatment].

    PubMed

    Tomura, Miki; Satoh, Koichi; Hanaoka, Mami; Tamura, Tetsuya; Kinouchi, Tomoya; Shinno, Kiyohito; Miyake, Hajimu; Niki, Hitoshi; Shoji, Akihiro

    2015-10-01

    A 35-year-old woman with a history of infertility, was presented to our hospital because of impaired consciousness and cerebellar ataxia, 14 days after in vitro fertilization. She received an embryo transfer under controlled ovarian hyper-stimulation. Magnetic resonance images revealed acute infarction in the cerebellum and brainstem. Magnetic resonance angiography showed a basilar artery occlusion at the end point. Following immediate intravenous rt-PA treatment, the symptoms disappeared completely. A transesophageal echocardiography revealed an atrial septal defect with a continuous left to right shunt. In addition, a Valsalva maneuver trans-esophageal echocardiography with injected saline showed the presence of jet bubbles in the left atrium crossing via the atrial septal defect. She was diagnosed with paradoxical cerebral embolism. Anticoagulant therapy was continued and she gave birth to a healthy baby. Deep vein thrombosis was associated with the ovarian hyper-stimulation syndrome that occurred during infertility treatment. As anti-phospholipid antibodies were weakly positive, the possibility of anti-phospholipid antibody syndrome was suggested. If a woman of childbearing age is presented because of stroke, it is important to administer initial therapy by keeping fertility in mind. Thrombolytic therapy for pregnant women should be carefully considered, because of the associated hazards; however, it is a very important treatment for maternal function after birth. PMID:26450079

  13. Random start or emergency IVF/in vitro maturation: a new rapid approach to fertility preservation.

    PubMed

    Robertson, David Mark; Gilchrist, Robert B; Ledger, William Leigh; Baerwald, Angela

    2016-06-01

    There is a need to develop rapid protocols for ovarian stimulation for women who wish to preserve their fertility following diagnosis of cancer. Conventional gonadotropin stimulation protocols are lengthy and are delayed until the start of the next menstrual period, potentially compromising cancer treatments. The development of random start IVF/in vitro maturation has made significant strides for enabling couples undergoing cancer therapy to achieve a family. However, several unanswered questions still remain. What do we know about the endocrinology of stimulating ovarian follicular activity outside the established protocols of stimulation during the follicular phase? This article explores what is known about antral follicle development during the menstrual cycle, novel ovarian stimulation proposals for optimizing assisted reproductive therapies in women, and direction. PMID:27248769

  14. Isolation, in vitro maturation, and fertilization of germinal vesicle oocytes obtained from the intact murine ovary.

    PubMed

    Randall, G W; Awadalla, S G; Shivers, C A

    1990-12-01

    The purpose of this investigation was to attempt to develop a process, utilizing a murine model, which would allow more efficient harvesting from the intact ovary and maturation in vitro of germinal vesicle (GV) oocytes. The recovery process yielded 25.5 +/- 4.5 (mean +/- SE) cumulus-free GV oocytes per animal. Treatment groups included culture medium (CM) supplemented with either estradiol (E2), follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), or prolactin (PRL). Among the hormone-free controls 83.2 +/- 1.6% of oocytes underwent GV breakdown, whereas 25.3 +/- 2.6% developed to the first polar body stage (PB-1) following 18 hr of incubation (n = 29 trials). Oocytes progressing to the PB-1 stage were inseminated in vitro. In vitro fertilization (IVF) of pooled in vitro matured (IVM) PB-1 oocytes (judged by two-cell formation) was 19.9%, which was significantly lower than in the group of in vivo matured oocytes (74.4%). E2 significantly increased the percentage of GV breakdown (control, 76.8 +/- 2.5%; E2 at 10 ng/ml, 92.9 +/- 2.5%, P less than 0.001; E2 at 100 ng/ml, 93.7 +/- 2.1%, P less than 0.001; and E2 at 1 micrograms/ml, 86.7 +/- 3.3%, P less than 0.05) but not PB-1 formation. Neither FSH nor hCG significantly increased GV breakdown or PB-1 formation.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. Tricho-rhino-phalangeal syndrome type 1 as an outcome of in vitro fertilization?

    PubMed

    Karaer, K; Yüksel, Z

    2014-01-01

    Trichorhinophalangeal syndrome type I [OMIM #190350] is an autosomal dominant disorder. Common features are: Slowly growing sparse hair, laterally thin eyebrows, bulbous tip of the nose, long philtrum, thin upper lip, protruding ears. Common skeletal anomalies include shortening of phalanges and metacarpals causing mild to severe brachydactyly, cone shaped epiphyses, hip dysplasia and short stature. Recently many reports have been published on the use of assisted reproductive technology (ART) and the increased risk of congenital major malformations or syndromes. We present a 6 years old Turkish Trichorhinophalangeal syndrome (TRPS) case of a twin pair after in vitro fertilization (IVF). TRPS with IVF pregnancy has not been reported previously. This new case reported herein will contribute to a better understanding whether ART pregnancy increases congenital malformations. PMID:24783650

  16. The In Vitro Fertilization Scheduler at the University of North Carolina and Its Satellite Clinics

    PubMed Central

    Cahill, Laurie; Bailey, Linda

    1988-01-01

    To facilitate scheduling of in vitro fertilization and gamete intrafallopian tube transfer (IVF-GIFT) patients, we developed a program (the IVF Scheduler) which uses the starting date of the last menstrual period, the number of days between starting dates of the last two menstrual periods (cycle length), and the month of requested treatment to calculate the date on which IVF-GIFT treatment should begin. Since cycle length is variable for each patient this calculation is performed for each of the three cycles prior to treatment so a multi-service, multi-site team can efficiently plan its operation. The program is menu-driven for ease of use. Benefits include allowing the nurse-coordinator to adjust treatment schedules easily; the development of the Scheduler into a pre-admission chart for nursing notes; and the delegation of primary scheduling duties from the nurse-coordinator to the IVF-GIFT secretary.

  17. Generating Transgenic Mice by Lentiviral Transduction of Spermatozoa Followed by In Vitro Fertilization and Embryo Transfer.

    PubMed

    Chandrashekran, Anil; Casimir, Colin; Dibb, Nick; Readhead, Carol; Winston, Robert

    2016-01-01

    Most transgenic technologies rely on the oocyte as a substrate for genetic modification. Transgenics animals are usually generated by the injection of the gene constructs (including lentiviruses encoding gene constructs or modified embryonic stem cells) into the pronucleus of a fertilized egg followed by the transfer of the injected embryos into the uterus of a foster mother. Male germ cells also have potential as templates for transgenic development. We have previously shown that mature sperm can be utilized as template for lentiviral transduction and as such used to generate transgenic mice efficiently with germ line capabilities. We provide here a detailed protocol that is relatively simple, to establish transgenic mice using lentivirally transduced spermatozoa. This protocol employs a well-established lentiviral gene delivery system (usual for somatic cells) delivering a variety of transgenes to be directly used with sperm, and the subsequent use of these modified sperm in in vitro fertilization studies and embryo transfer into foster female mice, for the establishment of transgenic mice. PMID:27317176

  18. Estradiol in saliva for monitoring follicular stimulation in an in vitro fertilization program

    SciTech Connect

    Belkien, L.D.; Bordt, J.; Moeller, P.; Hano, R.; Nieschlag, E.

    1985-09-01

    A rapid and sensitive radioimmunoassay (RIA) was developed to compare serum and saliva estradiol (E/sub 2/) levels in patients undergoing ovulation induction in an in vitro fertilization and embryo transfer (IVF-ET) program. Serum and saliva E/sub 2/ were compared in 23 patients. The sensitivity of the saliva RIA standard curve was 11 fmol/tube (equal to 3.2 pg/tube). There was a highly significant correlation between serum and saliva E/sub 2/ throughout the stimulated cycles. The ratio of serum to saliva E/sub 2/ was constant throughout the stimulated cycles. The E/sub 2/ concentration per follicle was 1548 pmol/l in serum and 23 pmol/l in saliva. Mean E/sub 2/ levels in saliva (+/- SD) were 74 +/- 21 pmol/l at midcycle and 46 +/- 12 pmol/l at midluteal phase. The findings indicate that measurement of saliva E/sub 2/ provides a reliable, noninvasive method and may replace serum measurements for monitoring stimulated cycles in an IVF-ET program.

  19. [Morphological changes in gametes of tiger barb Puntius tetrazona (Cypriniformes: Cyprinidae) and the implementation of in vitro fertilization].

    PubMed

    Domínguez-Castanedo, Omar; Toledano-Olivares, Ángel; Martínez-Espinosa, David; Ávalos-Rodríguez, Alejandro

    2014-12-01

    The production of ornamental fishes represents an economic activity of a growing number of Mexican families. Nevertheless, the reproduction of fish in captivity is one of the complications faced by farmers. This study was set up to: (i) evaluate the morphological and functional changes induced by hydration in the gametes of fish tiger barb (Puntius tetrazona; 240 samples) at tree times after hydration (10, 20 and 30s) with classic spermograms (volume, sperm concentration, viability, motility, and normal morphology); and (ii) evaluate the implementation of in vitro fertilization based on the ovulation rate, the percentage of fertilization and hatching; and the larval numbers obtained after 72 hours. The average volume of milt was 3.0 ± 0.7 μL, and the minimum, maximum and average concentration of sperm was 44.4 x 10(6) spz/mL, 52.3 x 10(6) spz/mL, and 48.1 ± 5.9 x 10(6) spz/mL, respectively. The viability and motility of the sperm was 84.6 ± 3.2% and 81.5 ± 2.2%, respectively. The diameter of the sperm with/without water contact was 2.10 ± 6 μm and 3.8 ± 1.0 μm (p < 0.05); the largest diameter was recorded 30 seconds after the contact with water. For oocytes, the smaller and larger diameters were recorded at 10 and 30s, respectively (both with/without water contact); the oocytes diameters after 10 and 30 seconds of contact with water were 1.11 and 1.55 mm, respectively. A higher ovulation rate was recorded using the in vitro fertilization: 250 ± 50 oocytes versus 28 ± 09 oocytes (during natural fertilization; p < 0.05). Nevertheless, fertilization and hatching rates were higher for the natural fertilization (80 and 60%, respectively). Considering the number of larvae obtained after 72 hours, our results showed a higher value for the in vitro fertilization (75 ± 18 compared to 13.4 ± 12 of the natural fertilization; p < 0.05). We propose this fish as a model for other ornamental fishes of commercial interest. Our results demonstrate that the in vitro

  20. [Morphological changes in gametes of tiger barb Puntius tetrazona (Cypriniformes: Cyprinidae) and the implementation of in vitro fertilization].

    PubMed

    Domínguez-Castanedo, Omar; Toledano-Olivares, Ángel; Martínez-Espinosa, David; Ávalos-Rodríguez, Alejandro

    2014-12-01

    The production of ornamental fishes represents an economic activity of a growing number of Mexican families. Nevertheless, the reproduction of fish in captivity is one of the complications faced by farmers. This study was set up to: (i) evaluate the morphological and functional changes induced by hydration in the gametes of fish tiger barb (Puntius tetrazona; 240 samples) at tree times after hydration (10, 20 and 30s) with classic spermograms (volume, sperm concentration, viability, motility, and normal morphology); and (ii) evaluate the implementation of in vitro fertilization based on the ovulation rate, the percentage of fertilization and hatching; and the larval numbers obtained after 72 hours. The average volume of milt was 3.0 ± 0.7 μL, and the minimum, maximum and average concentration of sperm was 44.4 x 10(6) spz/mL, 52.3 x 10(6) spz/mL, and 48.1 ± 5.9 x 10(6) spz/mL, respectively. The viability and motility of the sperm was 84.6 ± 3.2% and 81.5 ± 2.2%, respectively. The diameter of the sperm with/without water contact was 2.10 ± 6 μm and 3.8 ± 1.0 μm (p < 0.05); the largest diameter was recorded 30 seconds after the contact with water. For oocytes, the smaller and larger diameters were recorded at 10 and 30s, respectively (both with/without water contact); the oocytes diameters after 10 and 30 seconds of contact with water were 1.11 and 1.55 mm, respectively. A higher ovulation rate was recorded using the in vitro fertilization: 250 ± 50 oocytes versus 28 ± 09 oocytes (during natural fertilization; p < 0.05). Nevertheless, fertilization and hatching rates were higher for the natural fertilization (80 and 60%, respectively). Considering the number of larvae obtained after 72 hours, our results showed a higher value for the in vitro fertilization (75 ± 18 compared to 13.4 ± 12 of the natural fertilization; p < 0.05). We propose this fish as a model for other ornamental fishes of commercial interest. Our results demonstrate that the in vitro

  1. Effect of polyvinylpyrrolidone on bovine oocyte maturation in vitro and subsequent fertilization and embryonic development.

    PubMed

    Chung, Jin-Tae; Tosca, Lucie; Huang, Tian-Hua; Xu, Lan; Niwa, Koji; Chian, Ri-Cheng

    2007-08-01

    The exact role of polyvinylpyrrolidone (PVP) in culture medium for oocyte maturation is still largely unknown. Bovine cumulus-oocyte complexes (COC) were cultured in in-vitro maturation (IVM) medium supplemented with 10% fetal bovine serum (FBS), 0.3% PVP (K-90) or 10% serum substitute supplement (SSS) respectively. The rates of oocyte maturation, fertilization and early embryonic development were evaluated. In addition, the status of DNA fragmentation in the oocytes was determined by comet assay, and the ratio of trophectoderm (TE) cells and inner cell mass (ICM) in blastocysts was determined by differential staining. Furthermore, the percentage of apoptotic cells in the blastocysts was examined by TUNEL assay. The results indicated that the effect of PVP in IVM medium was similar to FBS in terms of oocyte maturation and subsequent embryonic development. However, the addition of SSS in IVM medium retarded further embryonic development and resulted in more oocyte DNA fragmentation and a higher ratio of TE cells and ICM in the blastocysts. However, the number of apoptotic cells in blastocysts was similar among the three groups. These results suggest for the first time that the addition of PVP in oocyte maturation medium is not only a suitable substitute for serum but is also beneficial to in-vitro oocyte maturation. PMID:17697497

  2. Recommendations for development of an emergency plan for in vitro fertilization programs: a committee opinion.

    PubMed

    2016-05-01

    All in vitro fertilization programs and clinics should have a plan to protect fresh and cryopreserved human tissue (embryos, oocytes, sperm) and to provide for continuation of patient care in the event of an emergency or natural disaster. This document was reviewed and affirmed by the Practice Committee in 2015.

  3. Bovine serum albumin (BSA) can replace patient serum as a protein source in an in vitro fertilization (IVF) program.

    PubMed

    Benadiva, C A; Kuczynski-Brown, B; Maguire, T G; Mastoianni, L; Flickinger, G L

    1989-06-01

    Alternate protein sources have been suggested to replace the commonly used cord or patient serum for in vitro fertilization (IVF) procedures. During an 11-month period 127 patients treated for in vitro fertilization had either their serum (N = 71) or bovine serum albumin (BSA; N = 56) used as the protein source in the insemination and growth media. Ham's F-10 + 0.5% BSA was used for sperm swim-up and insemination media and 1% BSA was used for the growth media. Patient's serum was added to Ham's F-10 culture media at concentrations of 7.5 and 15% for insemination and growth, respectively. Embryo transfer was performed with Ham's F-10 containing 90% maternal serum in both groups. Fertilization rate of 259 oocytes inseminated in medium containing patient's serum did not differ when compared with 200 oocytes inseminated in medium containing BSA. Likewise, rates of abnormal fertilization, cleavage, and pregnancy were similar in both groups. In a second experiment, 148 normally fertilized oocytes were transferred after 24 hr in culture to growth media containing two different concentrations of BSA (0.5 or 1%). Cleavage rates for the two groups were similar and the percentage of embryos developed to greater than or equal to 4 cells did not differ significantly. We conclude that a single concentration of BSA can safely be used to supplement culture media in human IVF with several practical and economical benefits.

  4. Handling of boar spermatozoa during and after flow cytometric sex-sorting process to improve their in vitro fertilizing ability.

    PubMed

    del Olmo, D; Parrilla, I; Gil, M A; Maside, C; Tarantini, T; Angel, M A; Roca, J; Martinez, E A; Vazquez, J M

    2013-09-01

    The objective of this study was to develop an adequate sperm handling protocol in order to obtain a sex-sorted sperm population with an optimal fertilizing ability. For this purpose, different aspects of the sorting procedure were examined. The effects of the high dilution rates (experiment 1), type of collection medium used (experiment 2), and sheath fluid composition (experiment 3) on sorted boar sperm quality and function were evaluated. Sperm quality was assessed by motility and viability tests, whereas sperm function was evaluated by an in vitro fertilization assay which determined the penetration and polyspermy rates as well as the mean number of sperm penetrating each oocyte. In experiment 1, the results obtained indicated that the high dilution rates did not cause a decrease either in the sperm quality parameters evaluated or the in vitro fertilization ability of spermatozoa. In experiment 2, although sperm quality was not affected, fertilizing ability was compromised after sorting, regardless of the collection medium that was used. In the experiment 3, all groups displayed adequate sperm quality values, but higher in vitro fertility parameters were obtained for spermatozoa sorted in presence of EDTA in the sheath fluid and egg yolk (EY) in the collection media when compared with those sorted in absence of these protective agents. No differences in penetration rates between unsorted highly diluted (control) and sorted sperm in the presence of EDTA and EY were observed. In conclusion, fertilizing ability was compromised in sex-sorted sperm. The addition of EDTA to sheath fluid and EY to collection medium improved boar sperm fertilizing ability, and both agents should be included as essential media components in future studies. PMID:23746874

  5. Handling of boar spermatozoa during and after flow cytometric sex-sorting process to improve their in vitro fertilizing ability.

    PubMed

    del Olmo, D; Parrilla, I; Gil, M A; Maside, C; Tarantini, T; Angel, M A; Roca, J; Martinez, E A; Vazquez, J M

    2013-09-01

    The objective of this study was to develop an adequate sperm handling protocol in order to obtain a sex-sorted sperm population with an optimal fertilizing ability. For this purpose, different aspects of the sorting procedure were examined. The effects of the high dilution rates (experiment 1), type of collection medium used (experiment 2), and sheath fluid composition (experiment 3) on sorted boar sperm quality and function were evaluated. Sperm quality was assessed by motility and viability tests, whereas sperm function was evaluated by an in vitro fertilization assay which determined the penetration and polyspermy rates as well as the mean number of sperm penetrating each oocyte. In experiment 1, the results obtained indicated that the high dilution rates did not cause a decrease either in the sperm quality parameters evaluated or the in vitro fertilization ability of spermatozoa. In experiment 2, although sperm quality was not affected, fertilizing ability was compromised after sorting, regardless of the collection medium that was used. In the experiment 3, all groups displayed adequate sperm quality values, but higher in vitro fertility parameters were obtained for spermatozoa sorted in presence of EDTA in the sheath fluid and egg yolk (EY) in the collection media when compared with those sorted in absence of these protective agents. No differences in penetration rates between unsorted highly diluted (control) and sorted sperm in the presence of EDTA and EY were observed. In conclusion, fertilizing ability was compromised in sex-sorted sperm. The addition of EDTA to sheath fluid and EY to collection medium improved boar sperm fertilizing ability, and both agents should be included as essential media components in future studies.

  6. In vitro fertilization (IVF) from low or high antral follicle count pubertal beef heifers using semi-defined culture conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Antral follicle counts (AFC) vary among pubertal beef heifers. Our objective was to compare the in vitro maturation and fertilization of oocytes collected from low and high AFC heifers. Previously we reported results using serum-based IVF media and in this study report results using semi-defined m...

  7. Influence of temperature stress on in vitro fertilization and heat shock protein synthesis in maize (Zea mays L. ) reproductive tissues

    SciTech Connect

    Dupuis, I.; Dumas, C. )

    1990-10-01

    This study was conducted to investigate the response of maize (Zea mays) male and female mature reproductive tissues to temperature stress. We have tested the fertilization abilities of the stressed spikelets and pollen using in vitro pollination-fertilization to determine their respective tolerance to stress. The synthesis of heat shock proteins (HSPs) was also analyzed in male and female tissues using electrophoresis of {sup 35}S-labeled proteins and fluorography, to establish a relationship between the physiological and molecular responses. Pollen, spikelets, and pollinated spikelets were exposed to selected temperatures (4, 28, 32, 36, or 40{degree}C) and tested using an in vitro fertilization system. The fertilization rate is highly reduced when pollinated spikelets are exposed to temperatures over 36{degree}C. When pollen and spikelets are exposed separately to temperature stress, the female tissues appear resistant to 4 hours of cold stress (4{degree}C) or heat stress (40{degree}C). Under heat shock conditions, the synthesis of a typical set of HSPs is induced in the female tissues. In contrast, the mature pollen is sensitive to heat stress and is responsible for the failure of fertilization at high temperatures. At the molecular level, no heat shock response is detected in the mature pollen.

  8. Monozygotic multiple gestation following in vitro fertilization: analysis of seven cases from Japan.

    PubMed

    Yanaihara, Atsushi; Yorimitsu, Takeshi; Motoyama, Hiroshi; Watanabe, Hideaki; Kawamura, Toshihiro

    2007-01-01

    We present a series of monozygous multiple gestations achieved following in vitro fertilization (IVF): one case of monochorionic triplet pregnancy and six cases of dizygotic triplet pregnancy. From September 2000 to December 2006, all patients achieving clinical pregnancy by ART were reviewed (n = 2433). A 37 year-old woman who delivered a healthy singleton after IVF returned two years later for FET, and a single blastocyst was transferred. This also resulted in pregnancy, but TV-USG revealed a single gestational sac with three distinct amniotic sacs, each containing a distinct fetal pole with cardiac activity. This pregnancy was electively terminated at nine weeks' gestation. An additional six cases of dizygotic triplets established after fresh embryo transfer (no ICSI or assisted hatching) are also described. Of these, one resulted in a miscarriage at eight weeks' gestation and five patients have an ongoing pregnancy. This case series suggests the incidence of dizygotic/monochorionic triplets following IVF is approximately 10 times higher than the expected rate in unassisted conceptions, and underscores the importance of a conservative approach to lower the number of embryos at transfer. The role of embryo transfer technique and in vitro culture media in the twinning process requires further study.

  9. In vitro fertilization (IVF) using semi-defined culture conditions from low or high antral follicle count pubertal beef heifers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers, AFC were determined on 106 heifers using transrectal ultrasonography. Ten heifers with the lowest AFC (avg. 13.2) and 10 heifers with the highest AFC (avg. 27.4) with ev...

  10. Human amniotic fluid for fertilization and culture of human embryos: results of clinical trials in human in vitro fertilization (IVF) programs.

    PubMed

    Gianaroli, L; Trounson, A; King, C; Ferraretti, A; Chiappazzo, L; Bafaro, G

    1989-08-01

    We report the outcome of clinical trials carried out in two IVF programs, comparing the use of human amniotic fluid (HAF) as a complete medium to Whittingham's T6 medium containing human serum (T6 + 10% HS) for egg incubation, insemination, embryo culture, and embryo transfer. There were no significant differences in the clinical trials between HAF used alone as a complete medium and T6 + 10% HS in fertilization rates of eggs, cleavage rates of embryos up to 48 hours in culture, pregnancy success rates after embryo replacement or the outcome of pregnancies. There was no advantage in using T6 + 10% HS for fertilization of eggs and HAF as a complete medium for embryo culture and transfer in any of the parameters examined. We conclude that HAF does not meet the complete requirements of human eggs and embryos in vitro and further developments of culture media are required to obtain embryo development equivalent to that in vivo.

  11. Normal fertilization in men with high antibody sperm binding by the addition of sufficient unbound sperm in vitro.

    PubMed

    Hamilton, F; Gutlay-Yeo, A L; Meldrum, D R

    1989-12-01

    A high incidence of fertilization failure has been reported in men with over 70% of their sperm bound with isoantibodies. In three men with greater than 80% antisperm antibody binding with IgG and IgA immunoglobulins, a normal rate of fertilization (29/46 oocytes; 63%) was achieved by adding a sufficient number of motile sperm to provide at least 50,000 unbound sperm per oocyte. This method appears to be simpler and more effective than attempting to separate unbound sperm in vitro.

  12. In vitro fertility rate of 129 strain is improved by Buserelin (GnRH) administration prior to superovulation

    PubMed Central

    Vasudevan, K; Sztein, J M

    2012-01-01

    The 129 mice are well recognized for their low fertility and it is speculated that this lack of fertility may be due to oocyte condition. In this study we investigated superovulation regimens for 129S1/SvImJ mouse strain to improve the oocyte quality and fertility rate of in vitro fertilization (IVF). Female mice were divided into four groups based on hormone and timing of injection. Group 1 received pregnant mare serum gonatotropin (PMSG) and 48 hours later human chorionic gonadotropin (hCG); using the same dose, group 2 received hCG 52 hours post PMSG and group 3, 55 hours post PMSG. Group 4 received Buserelin (gonadotropin releasing hormone agonist [GnRH]) followed 24 hours later by PMSG and then hCG 55 hours post PMSG. IVF was performed using 129S1/SvImJ oocytes and sperm; C57BL/6J sperm with 129S1/SvImJ oocytes was used as fertility control. The IVF fertility rate was 1% (Groups 1 & 2), 17% (Group 3) and 55% (Group 4) for 129 oocytes fertilized with 129 sperm. For 129 oocytes fertilized with C57BL/6J sperm, the fertility rate was 5% (Group 1) 10% (Group 2) 40% (Group 3) and 59% (Group 4).-These results suggest that extending the interval time between PMSG and hCG and giving GnRH in addition to the standard PMSG and hCG treatment can improve IVF fertility rate of 129S1/SvImJ strain mice significantly. PMID:23097563

  13. In vitro fertility rate of 129 strain is improved by buserelin (gonadotropin-releasing hormone) administration prior to superovulation.

    PubMed

    Vasudevan, K; Sztein, J M

    2012-10-01

    The 129 mice are well recognized for their low fertility and it is speculated that this lack of fertility may be due to the oocyte condition. In this study we investigated superovulation regimens for the 129S1/SvImJ mouse strain to improve the oocyte quality and fertility rate of in vitro fertilization (IVF). Female mice were divided into four groups based on hormone and timing of injection. Group 1 received pregnant mare serum gonadotropin (PMSG) and 48 h later human chorionic gonadotropin (hCG); using the same dose, group 2 received hCG 52 h post-PMSG and group 3, 55 h post-PMSG. Group 4 received buserelin (gonadotropin-releasing hormone agonist [GnRH]) followed 24 h later by PMSG and then hCG 55 h post-PMSG. IVF was performed using 129S1/SvImJ oocytes and sperm; C57BL/6J sperm with 129S1/SvImJ oocytes was used as fertility control. The IVF fertility rate was 1% (Groups 1 and 2), 17% (Group 3) and 55% (Group 4) for 129 oocytes fertilized with 129 sperm. For 129 oocytes fertilized with C57BL/6J sperm, the fertility rate was 5% (Group 1), 10% (Group 2), 40% (Group 3) and 59% (Group 4). These results suggest that extending the interval time between PMSG and hCG and giving GnRH in addition to the standard PMSG and hCG treatments can improve IVF fertility rate of 129S1/SvImJ mouse strains significantly.

  14. Improvement in in vitro fertilization outcome following in vivo synchronization of oocyte maturation in mice

    PubMed Central

    Muhsen-Alanssari, Saeeda A; Dees, WL; Ridha-Albarzanchi, Mundhir T; Kraemer, Duane C

    2015-01-01

    Synchronization of oocyte maturation in vitro has been shown to produce higher in vitro fertilization (IVF) rates than those observed in oocytes matured in vitro without synchronization. However, the increased IVF rates never exceeded those observed in oocytes matured in vivo without synchronization. This study was therefore designed to define the effect of in vivo synchronization of oocyte maturation on IVF rates. Mice were superovulated and orally treated with 7.5 mg cilostazol (CLZ), a phosphodiesterase 3A (PDE3A) inhibitor, to induce ovulation of immature oocytes at different stages depending on frequency and time of administration of CLZ. Mice treated with CLZ ovulated germinal vesicle (GV) or metaphase I (MI) oocytes that underwent maturation in vitro or in vivo (i.e. in the oviduct) followed by IVF. Superovulated control mice ovulated mature oocytes that underwent IVF directly upon collection. Ovulated MI oocytes matured in vitro or in vivo had similar maturation rates but significantly higher IVF rates, 2–4 cell embryos, than those observed in control oocytes. Ovulated GV oocytes matured in vitro showed similar maturation rates but significantly higher IVF rates than those observed in control oocytes. However, ovulated GV oocytes matured in vivo had significantly lower IVF rates than those noted in control oocytes. It is concluded that CLZ is able to synchronize oocyte maturation and improve IVF rates in superovulated mice. CLZ may be capable of showing similar effects in humans, especially since temporal arrest of human oocyte maturation with other PDE3A inhibitors in vitro was found to improve oocyte competence level. The capability of a clinically approved PDE3A inhibitor to improve oocyte fertilization rates in mice at doses extrapolated from human therapeutic doses suggests the potential scenario of the inclusion of CLZ in superovulation programs. This may improve IVF outcomes in infertile patients. PMID:25245076

  15. Rapid evaluation of soluble HLA-G levels in supernatants of in vitro fertilized embryos.

    PubMed

    Rebmann, Vera; Switala, Magdalena; Eue, Ines; Schwahn, Eva; Merzenich, Markus; Grosse-Wilde, Hans

    2007-04-01

    Human leukocyte antigen G (HLA-G) molecules are crucial for the maternal tolerance against the fetus during pregnancy. Thus, the presence of soluble HLA-G (sHLA-G) in embryo cultures is thought to be correlated to a successful pregnancy after assisted reproductive techniques (ART). Here, we established a rapid detection assay based on Luminex technology, which can be integrated into ART proceedings, allowing sHLA-G quantification in sample volumes of only 10 microl within 1.5 hours. Using this method, sHLA-G levels of 526 single-embryo cultures, 47 two-embryo cultures, and 15 three-embryo cultures were analyzed corresponding to 313 ART cycles. In 117 embryo cultures, sHLA-G was detectable. In single-embryo cultures, the sHLA-G levels were positively correlated to embryo quality (p = 0.048, r = 0.20, n = 100). The presence of sHLA-G in embryo cultures was significantly (p < 0.0001) associated with clinical pregnancy after intracytoplasmatic sperm injections (ICSI), especially in couples with male factor infertility, but not after in vitro fertilization (IVF) or in couples with female infertility. Importantly, in sHLA-G negative embryos, the abortion rate was increased threefold (p = 0.04). In conclusion, the results obtained by our novel method support strongly the diagnostic relevance of sHLA-G for predicting pregnancy outcome after ART. The ultimate conditions for this prediction have to be further investigated in a multicenter study.

  16. Economic value of in vitro fertilization in Ukraine, Belarus, and Kazakhstan

    PubMed Central

    Mandrik, Olena; Knies, Saskia; Severens, Johan L

    2015-01-01

    Background An economic value calculation was performed to estimate the lifetime net present value of in vitro fertilization (IVF) in Ukraine, Belarus, and Kazakhstan. Methods Net lifetime tax revenues were used to represent governmental benefits accruing from a hypothetical cohort of an IVF population born in 2009 using the methodology of generational accounting. Governmental expenses related to this population included social benefits, education and health care, unemployment support, and pensions. Where available, country-specific data referencing official sources were applied. Results The average health care cost needed to achieve one additional birth from the governmental perspective varied from $2,599 in Ukraine to $5,509 in Belarus. The net present value from the population born using IVF was positive in all countries: for Ukraine ($9,839), Belarus ($21,702), and Kazakhstan ($2,295). The break-even costs of drugs and supplies per IVF procedure is expected to be $3,870, $8,530, and $1,780, respectively. Probabilistic sensitivity analyses based on 5,000 simulations show that the average net present value per person remains positive: $1,894±$7,619, $27,925±$12,407, and $17,229±$24,637 in Ukraine, Belarus, and Kazakhstan, respectively. Conclusion Financing IVF may represent a good investment in terms of governmental financial returns, even in lower-income countries with state-financed health care systems such as Ukraine, Belarus, and Kazakhstan. PMID:26109873

  17. Comparison of different stimulation protocols used in in vitro fertilization: a review

    PubMed Central

    Shrestha, Deekshya; Feng, Huai L.

    2015-01-01

    Infertility is one of the major medical problems in the western world caused by genetic or epigenetic factors, or both, which has led to continuous research and advancements in the field of assisted reproductive technology (ART). Many stimulation protocols are available for controlled ovarian hyperstimulation (COH) in in vitro fertilization (IVF). This review compares the agonist long protocol, antagonist protocol and minimal stimulation protocol. Gonadotropin-releasing hormone (GnRH) antagonist and minimal stimulation protocol has shorter duration of treatment and less gonadotropin use. GnRH agonist long protocol is better in folliculogenesis and pregnancy rate, which is the imperative goal of COH. Despite its costly and lengthy approach, GnRH agonist long protocol has delivered satisfactory results in most women. On the other hand, patients with poor ovarian reserve may have greater advantage when considering minimal stimulation protocol. Evidently, it is crucial to have a larger scale studies with more focused comparisons, which take into account the differences in patients’ response criteria and additional confounding variables (age, BMI, previous IVF outcomes etc.), in order to reach to a more definite conclusions. PMID:26207230

  18. [Blood cell chimerism in dizygotic twins conceived by in vitro fertilization].

    PubMed

    Martos-Moreno, G Á; Campos, C; Flores, R; Yturriaga, R; Pérez-Jurado, L A; Argente, J

    2013-10-01

    We present a case of hematopoietic chimerism in dizygotic twins (male and female) conceived by in vitro fertilization (IVF). At 8 years of age a blood karyotype was performed on the female due to the presence of clitoromegaly. Two different lines: 46,XX (53%) and 46,XY (47%) were found. FISH studies confirmed the presence of the SRY gene in 46,XY cells. Karyotyping of the male showed two different lines: 46,XY (58%) and 46,XX (42%). SRY gene was present in 46,XY cells. Microsatellite analyses of blood DNA revealed tetra-allelic contribution at some autosomal loci with similar proportions of maternal and paternal alleles and X/Y chromosome dose. FISH in buccal mucous showed that all cells from the female were 46,XX and those from the male 46,XY. The gonadal karyotype in the female was 46,XX without SRY. Hence, we report 46,XX/46,XY chimerism in dizygotic twins. Blood chimerism was confirmed by performing FISH on the buccal cells of the patients.

  19. Tax credits, insurance, and in vitro fertilization in the U.S. military health care system.

    PubMed

    Wu, Mae; Henne, Melinda; Propst, Anthony

    2012-06-01

    The FAMILY Act, an income tax credit for infertility treatments, was introduced into the U.S. Senate on May 12, 2011. We estimated the costs and utilization of in vitro fertilization (IVF) in the military if infertility treatment became a tax credit or TRICARE benefit. We surveyed 7 military treatment facilities (MTFs) that offer IVF, with a 100% response rate. We first modeled the impact of the FAMILY Act on the MTFs. We then assessed the impact and costs of a TRICARE benefit for IVF. In 2009, MTFs performed 810 IVF cycles with average patient charges of $4961 and estimated pharmacy costs of $2K per cycle. With implementation of the FAMILY Act, we estimate an increase in IVF demand at the MTFs to 1165 annual cycles. With a TRICARE benefit, estimated demand would increase to 6,924 annual IVF cycles. MTF pharmacy costs would increase to $7.3 annually. TRICARE medical and pharmacy costs would exceed $24.4 million and $6.5 million, respectively. In conclusion, if the FAMILY Act becomes law, demand for IVF at MTFs will increase 29%, with a 50% decrease in patient medical expenses after tax credits. MTF pharmacy costs will rise, and additional staffing will be required to meet the demand. If IVF becomes a TRICARE benefit, demand for IVF will increase at least 2-fold. Current MTFs would be unable to absorb the increased demand, leading to increased TRICARE treatment costs at civilian centers. PMID:22730853

  20. In vitro oocyte maturation, fertilization and culture after ovum pick-up in an endangered gazelle (Gazella dama mhorr).

    PubMed

    Berlinguer, F; González, R; Succu, S; del Olmo, A; Garde, J J; Espeso, G; Gomendio, M; Ledda, S; Roldan, E R S

    2008-02-01

    The recovery of immature oocytes followed by in vitro maturation, fertilization and culture (IVMFC) allows the rescue of biological material of great genetic value for the establishment of genetic resource banks of endangered species. Studies exist on sperm cryopreservation of endangered Mohor gazelle (Gazella dama mhorr), but no work has been carried out yet on oocyte collection, fertilization and culture in this or related species. The purpose of this study was to develop a protocol for ovarian stimulation for the recovery of oocytes and subsequent IVMFC in the Mohor gazelle using frozen-thawed spermatozoa. Ovum pick-up was performed after ovarian stimulation with a total dose of 5.28 mg of ovine FSH. A total of 35 oocytes were recovered from 56 punctured follicles (62%) (N=6 females). Out of 29 cumulus-oocyte complexes matured in vitro, 3% were found at germinal vesicle stage, 7% at metaphase I, 21% were degenerated, and 69% advanced to metaphase II. Fertilization and cleavage rates of matured oocytes were 40 and 30%, respectively. Embryos cleaved in vitro up to the 6-8 cell stage but none progressed to the blastocyst stage, suggesting the existence of a developmental block and the need to improve culture conditions. Although more studies are needed to improve hormonal stimulation and oocyte harvesting, as well as IVMFC conditions, this study demonstrates for the first time the feasibility of in vitro fertilization with frozen-thawed semen of in vitro matured oocytes collected by ovum pick-up from FSH-stimulated endangered gazelles.

  1. In vitro oocyte maturation, fertilization and culture after ovum pick-up in an endangered gazelle (Gazella dama mhorr).

    PubMed

    Berlinguer, F; González, R; Succu, S; del Olmo, A; Garde, J J; Espeso, G; Gomendio, M; Ledda, S; Roldan, E R S

    2008-02-01

    The recovery of immature oocytes followed by in vitro maturation, fertilization and culture (IVMFC) allows the rescue of biological material of great genetic value for the establishment of genetic resource banks of endangered species. Studies exist on sperm cryopreservation of endangered Mohor gazelle (Gazella dama mhorr), but no work has been carried out yet on oocyte collection, fertilization and culture in this or related species. The purpose of this study was to develop a protocol for ovarian stimulation for the recovery of oocytes and subsequent IVMFC in the Mohor gazelle using frozen-thawed spermatozoa. Ovum pick-up was performed after ovarian stimulation with a total dose of 5.28 mg of ovine FSH. A total of 35 oocytes were recovered from 56 punctured follicles (62%) (N=6 females). Out of 29 cumulus-oocyte complexes matured in vitro, 3% were found at germinal vesicle stage, 7% at metaphase I, 21% were degenerated, and 69% advanced to metaphase II. Fertilization and cleavage rates of matured oocytes were 40 and 30%, respectively. Embryos cleaved in vitro up to the 6-8 cell stage but none progressed to the blastocyst stage, suggesting the existence of a developmental block and the need to improve culture conditions. Although more studies are needed to improve hormonal stimulation and oocyte harvesting, as well as IVMFC conditions, this study demonstrates for the first time the feasibility of in vitro fertilization with frozen-thawed semen of in vitro matured oocytes collected by ovum pick-up from FSH-stimulated endangered gazelles. PMID:18022681

  2. Promising system for selecting healthy in vitro-fertilized embryos in cattle.

    PubMed

    Sugimura, Satoshi; Akai, Tomonori; Hashiyada, Yutaka; Somfai, Tamás; Inaba, Yasushi; Hirayama, Muneyuki; Yamanouchi, Tadayuki; Matsuda, Hideo; Kobayashi, Shuji; Aikawa, Yoshio; Ohtake, Masaki; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei

    2012-01-01

    Conventionally, in vitro-fertilized (IVF) bovine embryos are morphologically evaluated at the time of embryo transfer to select those that are likely to establish a pregnancy. This method is, however, subjective and results in unreliable selection. Here we describe a novel selection system for IVF bovine blastocysts for transfer that traces the development of individual embryos with time-lapse cinematography in our developed microwell culture dish and analyzes embryonic metabolism. The system can noninvasively identify prognostic factors that reflect not only blastocyst qualities detected with histological, cytogenetic, and molecular analysis but also viability after transfer. By assessing a combination of identified prognostic factors--(i) timing of the first cleavage; (ii) number of blastomeres at the end of the first cleavage; (iii) presence or absence of multiple fragments at the end of the first cleavage; (iv) number of blastomeres at the onset of lag-phase, which results in temporary developmental arrest during the fourth or fifth cell cycle; and (v) oxygen consumption at the blastocyst stage--pregnancy success could be accurately predicted (78.9%). The conventional method or individual prognostic factors could not accurately predict pregnancy. No newborn calves showed neonatal overgrowth or death. Our results demonstrate that these five predictors and our system could provide objective and reliable selection of healthy IVF bovine embryos. PMID:22590579

  3. Degree and rate of growth discordance in dichorionic twins conceived by in vitro fertilization.

    PubMed

    Egic, Amira S; Mojovic, Donka V; Milovanovic, Zagorka M; Jurisic, Aleksandar B; Srbinovic, Ljubomir P; Krsmanovic, Suzana P; Karadzov-Orlic, Natasa T

    2014-01-01

    Objective. Our objective was to estimate degree and rate of discordant growth and its impact on perinatal outcome in dichorionic twin pregnancies conceived by in vitro fertilization (IVF) compared to those conceived spontaneously. Study Design. Growth discordance was defined as 90th percentiles for the study population. Adverse perinatal outcome was defined as 5-minute Apgar score <7 and/or admission to neonatal intensive care unit. Results. In the total study population of dichorionic twins (176 conceived by IVF and 215 spontaneously), 30% discordant growth represented the 90th percentile. After adjusting for gestational age, discordant twins conceived by IVF or spontaneously were at higher risk for adverse perinatal outcome (hazard ratio 4.4; 95% CI 2.4-8.3, P < 0.0001; hazard ratio 2.5; 95% CI 1.5-4.4, P = 0.001, resp.). Similar rates of 5-minute Apgar score <7, admission to neonatal intensive care unit, and delivery <34 weeks were found between discordant twins conceived by IVF and those conceived spontaneously. Conclusion. Dichorionic twins conceived by IVF are at similar risk for the rate and degree of discordant growth and adverse perinatal outcome compared to dichorionic twins conceived spontaneously.

  4. The impact of bariatric surgery on obesity-related infertility and in vitro fertilization outcomes.

    PubMed

    Tan, Orkun; Carr, Bruce R

    2012-12-01

    Obesity-related infertility is one of the most common problems of reproductive-age obese women who desire childbearing. The various types of bariatric surgeries have proved effective in controlling excessive weight gain, improving fertility, and preventing certain maternal and fetal complications in these women. This article summarizes the current evidence regarding the impact of bariatric surgery on obesity-related infertility and in vitro fertilization (IVF) outcomes. We have also attempted to draw conclusions about maternal and fetal risks and the benefits of bariatric surgery. Laparoscopic adjustable gastric banding and Roux-en-Y procedures are the two most commonly performed bariatric surgeries. Bariatric surgery was believed to improve menstrual irregularity and increase ovulation rate in anovulatory obese women, which lead to increased pregnancy rates. Although there are data in the literature suggesting the improvement of both the ovulatory function and the spontaneous pregnancy rates in obese women who lost weight after bariatric surgery, most of these are case-control studies with a small number of patients. The data are insufficient to determine an ideal time interval for pregnancy after bariatric surgery; however, the general consensus is that pregnancy should be delayed 12 to 18 months after bariatric surgery to avoid nutritional deficiencies. Few data exist regarding IVF success rates in women who have undergone bariatric surgery. One pairwise study discussed five patients who underwent bariatric surgery followed by IVF that resulted in three term pregnancies in three patients after the first IVF cycle. Many studies reported reductions in obesity-related pregnancy complications such as gestational diabetes and hypertensive disorders after bariatric surgery. Although data are inconsistent, some studies reported increased rate of preterm delivery and small for gestational age infants after bariatric surgery. Pregnancies after bariatric surgery may be

  5. A modified swim-up method reduces polyspermy during in vitro fertilization of porcine oocytes.

    PubMed

    Park, Chi-Hun; Lee, Sang-Goo; Choi, Don-Ho; Lee, Chang-Kyu

    2009-10-01

    The general method of porcine in vitro fertilization (IVF), involving the co-culture of both gametes in a medium drop, is thought to be the main reason for the high incidence of polyspermy. The aim of this study was to reduce the polyspermic fertilization of porcine embryos during IVF by the modified swim-up method, based on general sperm swim-up technique. Within this design, a 70 microm pore sized cell strainer was used to separate the sperm pellet placed at the bottom of a tube from the mature oocytes placed within the upper region. The separation of gametes using this permeable barrier was to ensure that only motile sperm gained access to the oocytes. It was found that the rate of polyspermy was significantly lowered for the sperm preparations from three boar breeds in modified swim-up method when compared with that of the general microdrop method (p<0.05). However, the penetration rates were found to be similar in both methods for two boar breeds. The average occurrence of blastocysts with more total cell number was higher in the modified swim-up method, while no significant difference in blastocyst rates between the two IVF methods was observed. The frequency of normal diploid embryos was also significantly higher in the modified swim-up method and polyploidy was more frequently observed in microdrop method (p<0.05). Our results demonstrated that the modified swim-up IVF method could reduce polyspermic penetration, and consequently produce better quality and karyotypically normal embryos in porcine IVF.

  6. Non-Invasive Assessment of Viability in Human Embryos Fertilized in Vitro

    PubMed Central

    Montskó, Gergely; Zrínyi, Zita; Farkas, Nelli; Várnagy, Ákos; Bódis, József

    2016-01-01

    Human reproduction is a relatively inefficient process and therefore the number of infertile couples is high. Assisted reproductive technologies (ART) have facilitated the birth of over five million children worldwide. ART, however, superimposes its own relative inefficiency on the preexisting inefficiency of normal reproduction. The efficiency (expressed as pregnancy rate) is generally not more than 30%. Modern reproductive medicine is gradually moving from multiple embryo transfer to the transfer of a single embryo, mainly because of obvious and unwanted side effects of multiple embryo transfer (e.g. „epidemic” multiple pregnancies). This concept, however, requires a fast, professional selection of the most viable embryo during the first few days of ART. Thus the aim of a modern ART is the safe transfer of a healthy, viable, single embryo. Accurate and rapid methods of quantifying embryo viability are needed to reach this goal. Methodological advances have the potential to make an important contribution, and there has been a drive to develop alternative non-invasive methods to better meet clinical needs. Metabolic and genetic profiling of spent embryo culture (SEC) media should offer an exceptional opportunity for the assessment of embryo viability. The current review focuses on the latest non-invasive diagnostic approaches for pre-implantation viability assessment of in vitro fertilized embryos. PMID:27683524

  7. Non-Invasive Assessment of Viability in Human Embryos Fertilized in Vitro.

    PubMed

    Kovács, Gábor L; Montskó, Gergely; Zrínyi, Zita; Farkas, Nelli; Várnagy, Ákos; Bódis, József

    2016-04-01

    Human reproduction is a relatively inefficient process and therefore the number of infertile couples is high. Assisted reproductive technologies (ART) have facilitated the birth of over five million children worldwide. ART, however, superimposes its own relative inefficiency on the preexisting inefficiency of normal reproduction. The efficiency (expressed as pregnancy rate) is generally not more than 30%. Modern reproductive medicine is gradually moving from multiple embryo transfer to the transfer of a single embryo, mainly because of obvious and unwanted side effects of multiple embryo transfer (e.g. "epidemic" multiple pregnancies). This concept, however, requires a fast, professional selection of the most viable embryo during the first few days of ART. Thus the aim of a modern ART is the safe transfer of a healthy, viable, single embryo. Accurate and rapid methods of quantifying embryo viability are needed to reach this goal. Methodological advances have the potential to make an important contribution, and there has been a drive to develop alternative non-invasive methods to better meet clinical needs. Metabolic and genetic profiling of spent embryo culture (SEC) media should offer an exceptional opportunity for the assessment of embryo viability. The current review focuses on the latest non-invasive diagnostic approaches for pre-implantation viability assessment of in vitro fertilized embryos. PMID:27683524

  8. Non-Invasive Assessment of Viability in Human Embryos Fertilized in Vitro

    PubMed Central

    Montskó, Gergely; Zrínyi, Zita; Farkas, Nelli; Várnagy, Ákos; Bódis, József

    2016-01-01

    Human reproduction is a relatively inefficient process and therefore the number of infertile couples is high. Assisted reproductive technologies (ART) have facilitated the birth of over five million children worldwide. ART, however, superimposes its own relative inefficiency on the preexisting inefficiency of normal reproduction. The efficiency (expressed as pregnancy rate) is generally not more than 30%. Modern reproductive medicine is gradually moving from multiple embryo transfer to the transfer of a single embryo, mainly because of obvious and unwanted side effects of multiple embryo transfer (e.g. „epidemic” multiple pregnancies). This concept, however, requires a fast, professional selection of the most viable embryo during the first few days of ART. Thus the aim of a modern ART is the safe transfer of a healthy, viable, single embryo. Accurate and rapid methods of quantifying embryo viability are needed to reach this goal. Methodological advances have the potential to make an important contribution, and there has been a drive to develop alternative non-invasive methods to better meet clinical needs. Metabolic and genetic profiling of spent embryo culture (SEC) media should offer an exceptional opportunity for the assessment of embryo viability. The current review focuses on the latest non-invasive diagnostic approaches for pre-implantation viability assessment of in vitro fertilized embryos.

  9. Does dilation and curettage versus expectant management for spontaneous abortion in patients undergoing in vitro fertilization affect subsequent endometrial development?

    PubMed

    Moon, Kimberly S; Richter, Kevin S; Levy, Michael J; Widra, Eric A

    2009-11-01

    In in vitro fertilization patients, treatment of spontaneous abortion with dilation and curettage (D&C) versus expectant management has no long-term effect on subsequent endometrial development, as measured by change in endometrial thickness. A transient reduction in endometrial thickness was found within the first 6 months after D&C, which is a novel finding, but it is likely to have little or no effect on pregnancy rates given the small absolute effect on endometrial thickness.

  10. Does dilation and curettage versus expectant management for spontaneous abortion in patients undergoing in vitro fertilization affect subsequent endometrial development?

    PubMed

    Moon, Kimberly S; Richter, Kevin S; Levy, Michael J; Widra, Eric A

    2009-11-01

    In in vitro fertilization patients, treatment of spontaneous abortion with dilation and curettage (D&C) versus expectant management has no long-term effect on subsequent endometrial development, as measured by change in endometrial thickness. A transient reduction in endometrial thickness was found within the first 6 months after D&C, which is a novel finding, but it is likely to have little or no effect on pregnancy rates given the small absolute effect on endometrial thickness. PMID:19560759

  11. Microheater as an alternative to lasers for in-vitro fertilization applications

    NASA Astrophysics Data System (ADS)

    Palanker, Daniel V.; Turovets, Igor; Glazer, Rima; Reubinoff, Benjamin E.; Hilman, Dalia; Lewis, Aaron

    1999-06-01

    During the last decade various lasers have been applied to drilling of the micrometer-sized holes in the zona pellucida of oocytes for in-vitro fertilization applications. In this paper we describe an alternative approach to laser instrumentation based on microfabricated device capable of precise drilling of uniform holes in the zona pellucida of oocytes. This device consists of a thin (1 micrometer) film microheater built on the tip of glass capillary with a diameter varying between a few to a few tens of micrometers. Duration of the pulse of heat produced by this microheater determines the spatial confinement of the heat wave in the surrounding liquid medium. We have demonstrated that gradual microdrilling of the zona pellucida can be accomplished using a series of pulses with duration of about 300 microseconds when the microheater was held in contact with the zona pellucida. Pulse energy applied to 20 micrometer tip was about 4 (mu) J. In vitro development and hatching of 127 micromanipulated embryos was compared to 103 non-drilled control embryos. The technique was found to be highly efficient in creating round, uniform, well defined holes with a smooth wall surface, matching the size of the heating source. The architecture of the surrounding zona pellucida was unaffected by the drilling, as demonstrated by scanning electron microscopy. Micromanipulated embryos presented no signs of thermal damage under light microscopy. The rate of blastocyst formation and hatching was similar in the micromanipulated and control groups. Following further testing in animal models, this methodology may be used as a cost- effective alternative to laser-based instrumentation in clinical applications such as assisted hatching and embryo biopsy.

  12. Perinatal and somatic growth properties of preterm babies born from spontaneous and in vitro fertilization multiple pregnancies

    PubMed Central

    Ramoğlu, Mehmet Gökhan; Kavuncuoğlu, Sultan; Özbek, Sibel; Aldemir, Esin

    2014-01-01

    Aim: The objective of this study was to examine perinatal and neonatal properties of preterm infants with a corrected age of 24–36 months who were born as a result of spontaneous and in vitro fertilization multiple pregnancies, to interrogate somatic growth properties of these infants and evaluate the factors which had an impact by comparing groups. Material and Methods: A total of 125 children with a birth weight below 2 500 g and a gestational age below the 37th gestational week 60 of whom were born as a result of in vitro fertilization multiple pregnancies and 65 of whom were born as a result of spontaneous multiple pregnancies were included in the study. Maternal age and morbidity, early rupture of membranes, birth weigth, gestational week, gender, APGAR score, hospitalization reasons in the neonatal period, requirement for intensive care, frequency of congenital anomaly, outpatient follow-up status, rehospitalization and socioeconomic levels were interrogated in the patients. Detailed physical examination and current height, weight and head circumference measurements were performed and the findings were placed in the growth curves of Neyzi et al. Ethics committee approval was received for this study from the ethics committee of Bakırköy Gynecology Obstetrics and Pediatrics Education and Research Hospital (12.10.2010; no:305). Results: The rate of cesarean section, mean maternal age, the rate of chronic disease in the mother and the rate of maternal disease which occured during pregnancy were significantly higher in the in vitro fertilization group (p<0.05). While no difference was found in mean gestational age, birth weight, rate of hospitalization, time of hospitalization, frequency of follow-up in the intensive care unit, rates of congenital anomaly and rehospitalization, APGAR score in the 5th minute was significantly higher in the in vitro fertilization group. The socioeconomical score was not different between the groups, but the in vitro

  13. Effects of pig follicular fluid on maturation of pig oocytes in vitro and on their subsequent fertilizing and developmental capacity in vitro.

    PubMed

    Yoshida, M; Ishizaki, Y; Kawagishi, H; Bamba, K; Kojima, Y

    1992-07-01

    This study examines the effects of pig follicular fluid on the maturation of pig oocytes and on their subsequent fertilizing and developmental capacity in vitro. The addition of pig follicular fluid or its fractions obtained by ultrafiltration, gel filtration and ion-exchange chromatography to maturation medium significantly increased the rates of nuclear maturation, normal fertilization and normal cleavage of pig oocytes after fertilization in vitro: the rates of normal fertilization and cleavage were 2-4 times higher than those in the control medium. The efficacy of pig follicular fluid was lost after heating at 56 degrees C for 30 min, whereas no significant decrease in activity was observed after defatting. In addition, the effective component(s) was partially purified by ultrafiltration, gel filtration and ion-exchange chromatography: the activity was observed in the fraction (UF2; M(r) 10,000-20,000) obtained by ultrafiltration. Activity was found in the first fraction (G1) obtained by gel filtration of UF2. Among three fractions obtained by ion-exchange chromatography of G1, only the third fraction had the activity. The results indicate that pig follicular fluid contains an acidic substance(s) (M(r) 10,000-200,000) that promotes oocyte maturation.

  14. Stress and stress-related hormones during in-vitro fertilization treatment.

    PubMed

    Harlow, C R; Fahy, U M; Talbot, W M; Wardle, P G; Hull, M G

    1996-02-01

    Whether stress and infertility are linked as cause or consequence is unclear, and there is no consensus on the most appropriate methods for measuring stress in infertile women. To address this question, we measured changes in biochemical and questionnaire-based assessments of stress in infertile women. Median baseline, follicular phase and pre-operative serum prolactin (229, 311 and 457 mIU/l) cortisol (278, 369 and 496 nmol/l) and state anxiety score (38, 40 and 49) respectively all increased during stimulated in-vitro fertilization (IVF) treatment. There was no such increase in a control group having similar laparoscopic surgery unrelated to infertility, or in women having unstimulated IVF without laparoscopy, suggesting that anxiety levels are greatest in stimulated IVF, increase as a result of the treatment, and are adequately reflected by state anxiety scores. Baseline serum prolactin in unstimulated IVF (384 mIU/l) was significantly higher than control (177 mIU/l), although this was not reflected in serum cortisol or state anxiety score. Trait anxiety was constant within and between groups, suggesting that stress is not contributing greatly to the infertility. Women who achieved a pregnancy had similar state anxiety scores to those who failed, suggesting that the degree of anxiety observed during IVF treatment is unlikely to influence the chance of pregnancy. There was a trend towards lower trait anxiety in women who became pregnant, but the numbers were small.

  15. Effect of body mass index and age on in vitro fertilization in polycystic ovary syndrome

    PubMed Central

    Kalem, Müberra Namlı; Kalem, Ziya; Sarı, Tamer; Ateş, Can; Gürgan, Timur

    2016-01-01

    Objective The aim of this study was to investigate age-related variations in the effect of body mass index (BMI) on in vitro fertilization (IVF) outcomes. Material and Methods This was a cohort study conducted by retrospectively investigating the IVF cycles of 653 polycystic ovary syndrome (PCOS) patients under the age of 40 years who were diagnosed based on the Rotterdam criteria in a private IVF clinic between 2005 and 2015. The study included data from 653 IVF cycles of PCOS patients. The patients were classified into three groups based on their BMI, i.e., normal weight (n=299), overweight (n=208), and obese (n=146). The patients were also grouped by age: 562 patients were under the age of 35 years and 91 patients were above the age of 35 years. Then, BMI- and age-related variations in the IVF cycle parameters and clinical pregnancy rates of patients with PCOS were investigated. The Mantel–Haenszel Chi-square statistical assessment method was used to determine whether the effect of BMI on IVF outcomes varies with age. Results Variations in cycle variables with BMI and age showed that IVF cycles were negatively affected by increases in obesity and age. Clinical pregnancy rates were found to be lower in the obese group than in the other groups, particularly in the age group above 35 years; however, this difference could not be proven statistically. Conclusion The present study evaluated obesity and clinical pregnancy rates in IVF cycles in PCOS patients according to age groups, and particularly in the obese group, the clinical pregnancy rates were observed to be lower in the age group ≥35 years than in the other BMI groups; however, this difference was found to be statistically insignificant. PMID:27403074

  16. Through the smoke: use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility.

    PubMed

    Camlin, Nicole J; McLaughlin, Eileen A; Holt, Janet E

    2014-12-15

    A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents' in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes.

  17. Early spontaneous multiple fetal pregnancy reduction is associated with adverse perinatal outcomes in in vitro fertilization cycles.

    PubMed

    Petrini, Allison C; Pereira, Nigel; Lekovich, Jovana P; Elias, Rony T; Spandorfer, Steven D

    2016-07-01

    The primary objective of this study is to investigate whether early spontaneous multiple fetal pregnancy reduction, also known as vanishing twin syndrome, is associated with adverse perinatal outcomes in fresh in vitro fertilization cycles. This is a retrospective cohort study of women with live singleton births with and without an early vanishing twin after fresh in vitro fertilization. Characteristics compared included incidence of preterm birth, overall birth weight, overall low birth weight, overall very low birth weight, and term low birth weight. In all, 4049 patients with live singleton births were included-853 and 3196 with and without a vanishing twin, respectively. The vanishing twin group had a lower overall birth weight compared to those without (3279.5 ± 369.9 vs 3368.6 ± 567.5 g; p < 0.01). Early vanishing twin was also associated with an increased odds of overall low birth weight (odds ratio: 1.75; 95% confidence interval: 1.36-2.25; p < 0.01) and increased odds of term low birth weight (odds ratio: 3.44; 95% confidence interval: 2.14-5.53; p < 0.01). Our study suggests that early vanishing twin is associated with lower overall birth weight and higher odds of overall low birth weight and term low birth weight in live singleton births after fresh in vitro fertilization. PMID:27638897

  18. Cryopreservation of human embryos and its contribution to in vitro fertilization success rates.

    PubMed

    Wong, Kai Mee; Mastenbroek, Sebastiaan; Repping, Sjoerd

    2014-07-01

    Cryopreservation of human embryos is now a routine procedure in assisted reproductive technologies laboratories. There is no consensus on the superiority of any protocol, and substantial differences exist among centers in day of embryo cryopreservation, freezing method, selection criteria for which embryos to freeze, method of embryo thawing, and endometrial preparation for transfer of frozen-thawed embryos. In the past decade, the number of frozen-thawed embryo transfer cycles per started in vitro fertilization (IVF) cycle increased steadily, and at the same time the percentage of frozen-thawed embryo transfers that resulted in live births increased. Currently, cryopreservation of human embryos is more important than ever for the cumulative pregnancy rate after IVF. Interestingly, success rates after frozen-thawed embryo transfer are now nearing the success rates of fresh embryo transfer. This supports the hypothesis of so called freeze-all strategies in IVF, in which all embryos are frozen and no fresh transfer is conducted, to optimize success rates. High-quality randomized controlled trials should be pursued to find out which cryopreservation protocol is best and whether the time has come to completely abandon fresh transfers.

  19. Impaired imprinted X chromosome inactivation is responsible for the skewed sex ratio following in vitro fertilization

    PubMed Central

    Tan, Kun; An, Lei; Miao, Kai; Ren, Likun; Hou, Zhuocheng; Tao, Li; Zhang, Zhenni; Wang, Xiaodong; Xia, Wei; Liu, Jinghao; Wang, Zhuqing; Xi, Guangyin; Gao, Shuai; Sui, Linlin; Zhu, De-Sheng; Wang, Shumin; Wu, Zhonghong; Bach, Ingolf; Chen, Dong-bao; Tian, Jianhui

    2016-01-01

    Dynamic epigenetic reprogramming occurs during normal embryonic development at the preimplantation stage. Erroneous epigenetic modifications due to environmental perturbations such as manipulation and culture of embryos during in vitro fertilization (IVF) are linked to various short- or long-term consequences. Among these, the skewed sex ratio, an indicator of reproductive hazards, was reported in bovine and porcine embryos and even human IVF newborns. However, since the first case of sex skewing reported in 1991, the underlying mechanisms remain unclear. We reported herein that sex ratio is skewed in mouse IVF offspring, and this was a result of female-biased peri-implantation developmental defects that were originated from impaired imprinted X chromosome inactivation (iXCI) through reduced ring finger protein 12 (Rnf12)/X-inactive specific transcript (Xist) expression. Compensation of impaired iXCI by overexpression of Rnf12 to up-regulate Xist significantly rescued female-biased developmental defects and corrected sex ratio in IVF offspring. Moreover, supplementation of an epigenetic modulator retinoic acid in embryo culture medium up-regulated Rnf12/Xist expression, improved iXCI, and successfully redeemed the skewed sex ratio to nearly 50% in mouse IVF offspring. Thus, our data show that iXCI is one of the major epigenetic barriers for the developmental competence of female embryos during preimplantation stage, and targeting erroneous epigenetic modifications may provide a potential approach for preventing IVF-associated complications. PMID:26951653

  20. Use of gonadotrophin-releasing hormone agonists in controlled ovarian hyperstimulation for in vitro fertilization.

    PubMed

    Muasher, S J

    1992-01-01

    The aim of ovarian hyperstimulation for in vitro fertilization (IVF) is the recruitment of multiple fertilizable healthy oocytes. Transfer of multiple embryos yields a better success rate than single-embryo transfers. Moreover, cryopreservation of excess pre-embryos allows patients an added opportunity to achieve a pregnancy without undergoing a repeat stimulated cycle. In the last 4 years, gonadotrophin-releasing hormone (Gn-RH) agonists have been used widely as adjuncts to gonadotrophins for ovarian hyperstimulation. Advantages of Gn-RH agonist use include prevention of a premature luteinising hormone (LH) surge, suppression of endogenous basal LH levels and recruitment of a larger cohort of follicles. Gn-RH agonists can be used in a long (suppression) or a short (stimulatory, flare-up) protocol. In our clinic, the use of Gn-RH agonist suppression (starting in the mid-luteal phase) prior to ovarian hyperstimulation was demonstrated to be extremely beneficial in intermediate and high responder patients but not in low responders (defined endocrinologically as patients with a basal follicle-stimulating hormone [FSH]: LH ratio of 1:1 and a basal LH:FSH ratio of greater than or equal to 1.5, respectively). We have not been able to demonstrate any beneficial effects from the use of Gn-RH agonist suppression in low responder patients (defined endocrinologically as patients with a basal FSH greater than or equal to 15 mIU/ml). In such low responder patients, the use of a 'flare-up' Gn-RH agonist protocol (Gn-RH agonist starting on day 2 of the cycle, followed by gonadotrophins on day 4 of the cycle), taking advantage of the initial agonistic stimulatory effect of Gn-RH agonists on endogenous FSH and LH secretion, has provided significant improvements in stimulation characteristics and better pregnancy results. It should be emphasised that comparisons of results cannot be attempted due to the selective use of each protocol in different patient populations.

  1. Alterations in uterine hemodynamics caused by uterine fibroids and their impact on in vitro fertilization outcomes

    PubMed Central

    Moon, Jei-Won; Kim, Jun-Bum; Kim, Sung-Hoon; Chae, Hee-Dong; Kang, Byung-Moon

    2015-01-01

    Objective To investigate the impact of fibroids on the blood flow of the uterine and subendometrial arteries and in vitro fertilization (IVF) outcomes. Methods In this study, we analyzed 86 IVF/intracytoplasmic sperm injection (ICSI) cycles in which a gonadotropin-releasing hormone antagonist protocol was used for controlled ovarian stimulation between January 2008 and March 2009. The subjects comprised 86 infertile women with (fibroid group, n=43) or without (control group, n=43) uterine fibroids. Results Patient characteristics were similar between the fibroid and control groups. The IVF/ICSI outcomes in patients with fibroids were similar to those of patients in the control group. The resistance index (RI) and pulsatile index (PI) of the uterine and subendometrial arteries on the day of embryo transfer were also comparable between the two groups. IVF outcomes and uterine hemodynamics in patients with multiple (≥2) fibroids were similar to those of patients with a single fibroid. However, clinical pregnancy and implantation rates were significantly lower in patients with fibroids who experienced uterine cavity distortion than in patients with fibroids who had a normal uterine cavity (both p<0.05). The RI and PI of the subendometrial artery were significantly higher on the day of embryo transfer in patients with fibroids who experienced uterine cavity distortion than in patients with fibroids who had a normal uterine cavity (both p<0.05). Conclusion Fibroids which distorting the uterine cavity might impair the subendometrial artery blood flow clinical pregnancy rate and embryo implantation rate in infertile patients undergoing IVF. Otherwise, IVF outcomes were not influenced by the presence of uterine fibroids. PMID:26816875

  2. Production of fertile offspring from oocytes grown in vitro by nuclear transfer in cattle.

    PubMed

    Hirao, Yuji; Naruse, Kenji; Kaneda, Masahiro; Somfai, Tamas; Iga, Kosuke; Shimizu, Manabu; Akagi, Satoshi; Cao, Feng; Kono, Tomohiro; Nagai, Takashi; Takenouchi, Naoki

    2013-09-01

    Because of recent advancements in reproductive technology, oocytes have attained an increasingly enriched value as a unique cell population in the production of offspring. The growing oocytes in the ovary are an immediate potential source that serve this need; however, complete oocyte growth before use is crucial. Our research objective was to create in vitro-grown (IVG) oocytes that would have the ability to perform specialized activities, including nuclear reprogramming, as an alternative to in vivo-grown oocytes. Bovine oocyte-granulosa cell complexes with a mean oocyte diameter of approximately 100 μm were cultured on Millicell membrane inserts, with culture medium supplemented with 4% polyvinylpyrrolidone (molecular weight, 360,000), 20 ng/ml androstenedione, 2 mM hypoxanthine, and 5 ng/ml bone morphogenetic protein 7. Oocyte viability after the 14-day culture period was 95%, and there was a 71% increase in oocyte volume. Upon induction of oocyte maturation, 61% of the IVG oocytes extruded a polar body. Eighty-four percent of the reconstructed IVG oocytes that used cumulus cells as donor cells underwent cleavage, and half of them became blastocysts. DNA methylation analyses of the satellite I and II regions of the blastocysts revealed a similar highly methylated status in the cloned embryos derived from in vivo-grown and IVG oocytes. Finally, one of the nine embryos reconstructed from the IVG oocytes developed into a living calf following embryo transfer. Fertility of the offspring was confirmed. In conclusion, the potential of a proportion of the IVG oocytes was comparable to that of in vivo-grown oocytes.

  3. Monitoring in-vitro bovine embryo development during the first days after fertilization (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Kandel, Mikhail E.; Rubessa, Marcello; Fernandes, Daniel; Nguyen, Tan H.; Wheeler, Matthew B.; Popescu, Gabriel

    2016-03-01

    Conventional label-based contrast enhancement techniques (e.g., fluorescence) frequently modify the genetic makeup of tagged cells, making them poor candidates for use in in-vitro fertilization applications. Instead, we choose a label-free form of contrast, based on interferometric imaging, sensitive to optical path length differences. Compared to, single HeLa cells, typical mammalian ova and embryos are more than an order of magnitude thicker. As a result, regions of large phase variation lead to phase wrapping and an overall reduction in signal intensity occurs due to multiple scattering. These effects manifest themselves in low-spatial frequencies (blurs), with the desired details buried in the background. We present a phase shifting interferometer that yields the derivative of the phase, a quantity whose value is particularly sensitive to local variations and fine details. We demonstrate that our new real-time imaging platform is valuable in measuring the multiday development of bovine embryos. Reconstructing the derivative of the image phase and amplitude, we characterize the motion of previously low-contrast structures, which are relevant for embryo viability tests.

  4. Factors Associated with the Success of In Vitro Fertilization in Women with Inflammatory Bowel Disease

    PubMed Central

    Oza, Sveta Shah; Pabby, Vikas; Dodge, Laura E.; Hacker, Michele R.; Fox, Janis H.; Moragianni, Vasiliki A.; Correia, Katharine; Missmer, Stacey A.; Ibrahim, Yetunde; Penzias, Alan S.; Burakoff, Robert; Friedman, Sonia

    2016-01-01

    Background It is unknown whether certain factors are associated with the success of in vitro fertilization (IVF) in women with inflammatory bowel disease (IBD). Aim This study assessed whether certain characteristics are associated with greater success of live birth following IVF. Methods In a cohort study of 8684 women with IBD seen at two tertiary care centers, we identified 121 women with IBD who underwent IVF. We assessed the effect of numerous factors on likelihood of achieving live birth after IVF. Results Seventy-one patients with ulcerative colitis (UC) and 49 patients with Crohn's disease (CD) were analyzed. Patients with UC who achieved a live birth were younger (p = 0.03), had a shorter duration of disease (p = 0.01), and were more likely to be in remission (p = 0.03) versus those who did not achieve live birth. Patients with CD who achieved live birth were younger (p < 0.001), had lower body mass index (BMI) (p = 0.02), and had lower cycle day 3 follicle-stimulating hormone levels (p = 0.02). There was no difference in likelihood of achieving live birth among patients in remission and those with mild or unknown disease status (p = 0.69), though most CD patients (79.5 %) were in remission. Prior surgery was not associated with live birth in patients with UC (p = 0.31) or CD (p = 0.62). Conclusions As in the general infertility population, younger patients and those with lower BMI were more likely to achieve live birth. History of surgery was not associated with live birth among IBD patients. This is important information for practitioners counseling IBD patients. PMID:26888767

  5. Suicidal risk among infertile women undergoing in-vitro fertilization: Incidence and risk factors.

    PubMed

    Shani, Chen; Yelena, Stukalina; Reut, Ben Kimhy; Adrian, Shulman; Sami, Hamdan

    2016-06-30

    Despite the fact that depression and other emotional distress are well documented in infertile women, little is known about the relationship between infertility and suicidal risk. The aim of this cross sectional study was to examine the rate of suicide risk (suicidal ideation/suicidal attempts) among 106 infertile women visiting Infertility and In-Vitro Fertilization (IVF) Hospital Unit, and to identify the demographic, medical and clinical correlates to suicidal risk. The incidence of suicide risk was 9.4%. Suicidal women were more likely to be childless or had fewer children and experienced higher levels of depressive symptoms. In addition, they reported more frequently on denial, social withdrawal and self-blame coping strategies compared to participants without suicidal risk. A multiple logistic regression model revealed that being childless, using non-positive reappraisal and exhibiting depressive symptoms were significant predictors of suicide risk in the future. These results suggest that routine assessment of suicidal risk and depression should be provided for infertile women in the course of IVF. Furthermore, future interventions should focus on helping them acquire different emotions regulation strategies and provide alternative skills for positive coping.

  6. HYPERGLYCOSYLATED HUMAN CHORIONIC GONADOTROPIN AS AN EARLY PREDICTOR OF PREGNANCY OUTCOMES AFTER IN VITRO FERTILIZATION

    PubMed Central

    Chuan, Sandy; Homer, Michael; Pandian, Raj; Conway, Deirdre; Garzo, Gabriel; Yeo, Lisa; Su, H. Irene

    2014-01-01

    Objective To determine if hyperglycosylated hCG (hhCG), produced by invasive trophoblasts, measured as early as 9 days after egg retrieval can predict ongoing pregnancies (OP) after in vitro fertilization and fresh embryo transfer (IVF-ET). Design Cohort Setting Academic ART center Patients Consecutive patients undergoing IVF-ET Interventions Serum hhCG and hCG levels measured 9 (D9) and 16 (D16) days after egg retrieval Outcome Ongoing pregnancy (OP) beyond 9 weeks of gestation Results OP (62 of 112 participants) was associated with higher D9 levels of hhCG and hCG However, hhCG was detectable in all D9 OP samples, while hCG was detectable in only 22%. D9 hhCG levels >110 pg/mL was 96% specific for OP, yielding a positive predictive value of 95%. Compared to D9 hCG levels, hhCG was more sensitive and had a larger area under the curve (0.87 vs. 0.67). Diagnostic test characteristics were similar between D16 hhCG and hCG levels. Conclusions In patients undergoing assisted reproduction, a test to detect pregnancy early and predict outcomes is highly desirable. HhCG is detectable in serum 9 days after egg retrieval IVF-ET cycles. At this early assessment, hhCG is superior to traditional hCG and highly predictive of ongoing pregnancies. PMID:24355054

  7. Support through patient internet-communities: Lived experience of Russian in vitro fertilization patients

    PubMed Central

    Isupova, Olga G.

    2011-01-01

    The article is concerned with the life experiences of infertile women going through infertility treatment and their need for social and psychological support, which they try to find in their immediate social environment. The Internet has become one place where everyone can find “people like oneself.” The best support is received from these people who are in the same life situation and are able and willing to share their lived experiences with each other. Communication via the Internet and the formation of a virtual community of patients has both positive and negative aspects, all of which are examined in the article. On the one hand, it creates a psychologically favorable atmosphere and might potentially increase the success rate of IVF treatment. On the other, this leads to the seclusion of patients within the circle of “similar people” and sometimes to negative attitudes towards people outside the circle. The article is based on the author's “netnography” research of a virtual community of Russian In-Vitro Fertilization (IVF)1 patients. PMID:21760835

  8. Development of a Chinese childbearing attitude questionnaire for infertile women receiving in vitro fertilization treatment.

    PubMed

    Lee, Shu-Hsin; Kuo, Ching-Pyng; Hsiao, Chiu-Yueh; Lu, Yen-Chiao; Hsu, Ming-Yi; Kuo, Pi-Chao; Lee, Maw-Shen; Lee, Meng-Chih

    2013-04-01

    The purpose of this study was to report the second phase of instrument development, a culturally sensitive questionnaire of childbearing attitudes to assess the psychosocial responses of infertile women. Using a nonexperimental quantitative design, we investigated 238 women who are undergoing in vitro fertilization treatment. Data collection and relevant planning occurred in two phases: in-depth interviews of women to generate items for the questionnaire and establishing the questionnaire's content and construct validity. Through factor analysis, five factors were extracted from the "attitude toward childbearing questionnaire": gender identification with self and society, insurance of marriage and inheritance, happy family life, spiritual investment, and continuing the family line and procreation. The total variance of these five factors was 64.31%. Cronbach's α and test-retest reliability were between .72 and .87 and between .60 and .76, respectively, demonstrating acceptable internal consistency and stability. The information obtained through the questionnaire could be used to provide infertile women with personal counseling and appropriate psychological support during and after assisted reproductive technology. PMID:23460456

  9. Recovery after unsuccessful in vitro fertilization: the complex role of resilience and marital relationships.

    PubMed

    Chochovski, Julijana; Moss, Simon A; Charman, Denise P

    2013-09-01

    The failure rate of in vitro fertilization (IVF) is around 75% per cycle. These unsuccessful attempts can provoke acute clinical depression and other problems. Although practitioners often recommend cognitive reappraisal, rather than avoidance, to cope with these difficulties, previous research has not established the psychosocial determinants of adaptive coping strategies. Arguably, resilience could encourage cognitive reappraisal, because resilient individuals feel confident they can overcome their emotions, whereas marital quality could prevent avoidance, because individuals feel secure enough to reflect upon their distress. Consequently, resilience and marital quality could facilitate recovery over time. To explore these possibilities, 184 women, all of whom had unsuccessfully completed IVF treatment, completed a questionnaire that gauged their levels of self-reported depression since their last IVF attempt as well as resilience and marital quality. Immediately after the unsuccessful attempt, resilience was inversely, whereas marital quality was positively, related to depression. However, within this cross-sectional sample after greater time had elapsed, marital quality became increasingly beneficial and was negatively associated with depression. These findings imply that resilience can curb the initial distress; in contrast, marital quality may enable individuals to reflect upon their trauma, initially amplifying distress but eventually facilitating recovery. Future research would benefit from longitudinal studies, illustrating whether resilience and marital quality at one time predict changes in distress at subsequent times.

  10. Risk Factors and Early Predictors for Heterotopic Pregnancy after In Vitro Fertilization

    PubMed Central

    Geng, Ling; Xia, Mingdi; Zhai, Junyu; Zhang, Wei; Zhang, Yuchao; Sun, Yinhua; Zhang, Jiangtao; Zhu, Dongyi; Zhao, Han; Chen, Zi-Jiang

    2015-01-01

    This study investigated the risk factors and early predictors for heterotopic pregnancy (HP) after in vitro fertilization and embryo transfer (IVF-ET). From January 2008 to January 2013, 41 cases of HP and 72 cases of intrauterine twin pregnancy after IVF-ET were recruited and retrospectively analyzed. Compared with intrauterine twin pregnancy group, the HP group had a lower basal luteinizing hormone (LH) level (P = 0.005) and more cases had a history of hydrosalpinx (P = 0.008). After 14 days of IVF-ET, the serum β-HCG (β-human chorionic gonadotropin), E2 (Estradiol) and P (Progesterone) levels were lower in HP group (P<0.001, respectively). Moreover, vaginal bleeding and abdominal pain were the significant features of HP before diagnosis (P<0.001, respectively). Further by logistic regression, serum β-hCG, P levels on the 14th day after ET, and vaginal bleeding were identified as the independent factors of HP. These results indicate that when two or more embryos transferred in IVF procedure, β-hCG, P levels on the 14th day after ET, and vaginal bleeding could be taken as predictors for HP. PMID:26510008

  11. Anesthesia Related Toxic Effects on In Vitro Fertilization Outcome: Burden of Proof.

    PubMed

    Matsota, Paraskevi; Kaminioti, Eva; Kostopanagiotou, Georgia

    2015-01-01

    Management of pain and anxiety during oocyte retrieval makes anesthesia an important part of the in vitro fertilization (IVF) procedures. There are many studies investigating the influence of anesthesia on IVF success. This review article provides an overview of published data regarding the potential toxic effects of different anesthetic techniques (Loco-regional, general anesthesia (GA), and monitored anesthesia care (MAC)), different anesthetic agents, and alternative medicine approach (principally acupuncture) on the IVF outcome. From our analysis, evidence of serious toxicity in humans is not well established. Trials regarding different anesthetic techniques ended up without clear conclusions. Studies about GA came up with conflicting results. A few trials relate GA with lower pregnancy rates, although some others failed to prove this conclusion. Furthermore, detectable amounts of some anesthetic agents are measurable in the follicular fluid but these findings are not strongly associated with toxicity. MAC and Loco-regional anesthesia appear as safe alternative choices and there is evidence of improved outcome. Whereas acupuncture may provide assistance increasing IVF success according to some trials, some others could not obtain these effects. Questions about the appropriate time of application and the underlying mechanism of action are not answered yet, so further investigation should be done. PMID:26161404

  12. Anesthesia Related Toxic Effects on In Vitro Fertilization Outcome: Burden of Proof

    PubMed Central

    Matsota, Paraskevi; Kaminioti, Eva; Kostopanagiotou, Georgia

    2015-01-01

    Management of pain and anxiety during oocyte retrieval makes anesthesia an important part of the in vitro fertilization (IVF) procedures. There are many studies investigating the influence of anesthesia on IVF success. This review article provides an overview of published data regarding the potential toxic effects of different anesthetic techniques (Loco-regional, general anesthesia (GA), and monitored anesthesia care (MAC)), different anesthetic agents, and alternative medicine approach (principally acupuncture) on the IVF outcome. From our analysis, evidence of serious toxicity in humans is not well established. Trials regarding different anesthetic techniques ended up without clear conclusions. Studies about GA came up with conflicting results. A few trials relate GA with lower pregnancy rates, although some others failed to prove this conclusion. Furthermore, detectable amounts of some anesthetic agents are measurable in the follicular fluid but these findings are not strongly associated with toxicity. MAC and Loco-regional anesthesia appear as safe alternative choices and there is evidence of improved outcome. Whereas acupuncture may provide assistance increasing IVF success according to some trials, some others could not obtain these effects. Questions about the appropriate time of application and the underlying mechanism of action are not answered yet, so further investigation should be done. PMID:26161404

  13. Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process.

    PubMed

    Scalici, E; Traver, S; Mullet, T; Molinari, N; Ferrières, A; Brunet, C; Belloc, S; Hamamah, S

    2016-01-01

    Circulating or "extracellular" microRNAs (miRNAs) detected in biological fluids, could be used as potential diagnostic and prognostic biomarkers of several disease, such as cancer, gynecological and pregnancy disorders. However, their contributions in female infertility and in vitro fertilization (IVF) remain unknown. This study investigated the expression profiles of five circulating miRNAs (let-7b, miR-29a, miR-30a, miR-140 and miR-320a) in human follicular fluid from 91 women with normal ovarian reserve and 30 with polycystic ovary syndrome (PCOS) and their ability to predict IVF outcomes. The combination of FF miR-30a, miR-140 and let-7b expression levels discriminated between PCOS and normal ovarian reserve with a specificity of 83.8% and a sensitivity of 70% (area under the ROC curve, AUC = 0.83 [0.73-0.92]; p < 0.0001). FF samples related to low number of mature oocytes (≤2) contained significant less miR-320a levels than those related to a number of mature oocytes >2 (p = 0.04). Moreover, FF let-7b predicted the development of expanded blastocysts with 70% sensitivity and 64.3% specificity (AUC = 0.67 [0.54-0.79]; p = 0.02) and FF miR-29a potential to predict clinical pregnancy outcome reached 0.68 [0.55-0.79] with a sensitivity of 83.3% and a specificity of 53.5% (p = 0.01). Therefore, these miRNAs could provide new helpful biomarkers to facilitate personalized medical care during IVF. PMID:27102646

  14. Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process

    PubMed Central

    Scalici, E.; Traver, S.; Mullet, T.; Molinari, N.; Ferrières, A.; Brunet, C.; Belloc, S.; Hamamah, S.

    2016-01-01

    Circulating or “extracellular” microRNAs (miRNAs) detected in biological fluids, could be used as potential diagnostic and prognostic biomarkers of several disease, such as cancer, gynecological and pregnancy disorders. However, their contributions in female infertility and in vitro fertilization (IVF) remain unknown. This study investigated the expression profiles of five circulating miRNAs (let-7b, miR-29a, miR-30a, miR-140 and miR-320a) in human follicular fluid from 91 women with normal ovarian reserve and 30 with polycystic ovary syndrome (PCOS) and their ability to predict IVF outcomes. The combination of FF miR-30a, miR-140 and let-7b expression levels discriminated between PCOS and normal ovarian reserve with a specificity of 83.8% and a sensitivity of 70% (area under the ROC curve, AUC = 0.83 [0.73–0.92]; p < 0.0001). FF samples related to low number of mature oocytes (≤2) contained significant less miR-320a levels than those related to a number of mature oocytes >2 (p = 0.04). Moreover, FF let-7b predicted the development of expanded blastocysts with 70% sensitivity and 64.3% specificity (AUC = 0.67 [0.54–0.79]; p = 0.02) and FF miR-29a potential to predict clinical pregnancy outcome reached 0.68 [0.55–0.79] with a sensitivity of 83.3% and a specificity of 53.5% (p = 0.01). Therefore, these miRNAs could provide new helpful biomarkers to facilitate personalized medical care during IVF. PMID:27102646

  15. Transforming hope: the lived experience of infertile women who terminated treatment after in vitro fertilization failure.

    PubMed

    Su, Tsann-Juu; Chen, Yueh-Chih

    2006-03-01

    Assisted reproductive treatments provide the hope of pregnancy for infertile women, but do not always turn this hope into reality. The purpose of this study was to explore the lived experience of infertile women who terminated treatment after in vitro fertilization (IVF) failure. Using a qualitative research design, 24 subjects were recruited who had experienced IVF failure and decided to terminate their treatment. Data were collected through interviews, and analyzed using interpretive research strategies of phenomenology. This study protocol was reviewed and approved by the University Review Board for Research. Informed consent was obtained from each subject. The theme of lived experience which emerged from the data was "transforming hope". This theme included three categories: (1) accepting the reality of infertility, (2) acknowledging the limitations of treatment involving high technology, and (3) re-identifying one's future. The results illustrated that counseling for these women should involve the provision of both positive and negative information, evaluation of the response to treatment, and assistance in defining their future.

  16. Grief responses and coping strategies among infertile women after failed in vitro fertilization treatment.

    PubMed

    Lee, Shu-Hsin; Wang, Shu-Chuan; Kuo, Ching-Pyng; Kuo, Pi-Chao; Lee, Maw-Sheng; Lee, Meng-Chih

    2010-09-01

    Reproductive technology has increased the childbearing potential for many infertile women, but in vitro fertilization (IVF) failures are common, which often trigger grief responses and coping strategies to manage the stressful life event. The present cross-sectional study investigated 66 women who had experienced at least one failure with IVF treatment. The data were gathered by a self-administered structured questionnaire, and included the participant's personal profile, grief responses and the Jalowiec's coping scale. The most common grief response among the respondents was bargaining, followed by acceptance, depression, anger, denial, and isolation. The order of coping strategies used, from highest-to-lowest, were confrontative, optimistic, self-reliant, fatalistic, supportive, evasive, palliative, and emotive. Use and self-perceived effectiveness among all coping strategies had a high correlation, except emotion. Bargaining, the most common grief response, was associated with a variety of coping strategies. All coping strategies were correlated with grief responses. The results of identifying the grief responses and associated coping strategies of women who have undergone failed IVF treatment may assist nurses and other health care professionals in their efforts to provide appropriate information, care and psychological support.

  17. Cryopreservation/transplantation of ovarian tissue and in vitro maturation of follicles and oocytes: Challenges for fertility preservation

    PubMed Central

    Varghese, Alex C; du Plessis, Stefan S; Falcone, Tommaso; Agarwal, Ashok

    2008-01-01

    Cryopreservation of ovarian tissue and in vitro follicle maturation are two emerging techniques for fertility preservation, especially in cancer patients. These treatment regimes are opening up more options and allow for more suitable choices to preserve fertility according to the patient's specific circumstances. If these technologies are to become widely accepted, they need to be safe, easy to perform and must obtain favorable results. The generation of healthy eggs with the normal genetic complement and the ability to develop into viable and healthy embryos requires tight regulation of oocyte development and maturation. Novel freezing techniques such as vitrification, along with whole ovary cryopreservation and three-dimensional follicle cultures, have shown favorable outcomes. The scope of this article is to take a comprehensively look at the challenges still faced in order for these novel technologies to be routinely employed with the aim of successful fertility preservation. PMID:18828928

  18. MEN’S BODY MASS INDEX IN RELATION TO EMBRYO QUALITY AND CLINICAL OUTCOMES IN COUPLES UNDERGOING IN VITRO FERTILIZATION

    PubMed Central

    Colaci, Daniela S.; Afeiche, Myriam; Gaskins, Audrey J.; Wright, Diane L.; Toth, Thomas L.; Tanrikut, Cigdem; Hauser, Russ; Chavarro, Jorge E.

    2012-01-01

    Objective To evaluate the association between men’s body mass index (BMI), early embryo quality and clinical outcomes in couples undergoing in vitro fertilization. Design Prospective cohort study. Setting Fertility clinic in an academic medical center. Patients 114 couples that underwent 172 ART cycles. Interventions None Main outcome measure Fertilization rate, embryo quality, implantation rate, clinical pregnancy rate and live birth rate. Results Fertilization rate was higher among obese men than among normal weight men (p-trend=0.04) in conventional IVF cycles. No significant associations were found between men’s BMI and the proportion of poor quality embryos on day 3 (p-trend =0.67), slow embryo cleavage rate (p-trend=0.17), or accelerated embryo cleavage rate (p-trend =0.07). Men’s BMI was unrelated to positive β-hCG rate (p-trend =0.37), clinical pregnancy rate (p-trend =0.91) or live birth rate (p-trend =0.42) per embryo transfer. Among couples undergoing ICSI, the odds of live birth in couples with obese male partners was 84% (95% CI 10%–97%) lower than the odds in couples with men with normal BMI (p-trend=0.04). Conclusion Our data suggest a possible deleterious effect of male obesity on the odds of having a live birth among couples undergoing ICSI. PMID:22884013

  19. Evaluation of a synthetic serum substitute to replace fetal cord serum for human oocyte fertilization and embryo growth in vitro.

    PubMed

    Psalti, I; Loumaye, E; Pensis, M; Depreester, S; Thomas, K

    1989-11-01

    A comparison was made between a serum substitute. UltroSer G (Gibco, Ghent, Belgium) (2%) (medium B) and 10% human fetal cord serum (medium A), as regards their ability to support 1-cell and 2-cell mice embryo development in vitro. Sixty percent and 56% of the 1-cell embryos reached the expanded blastocyst stage when cultured in media A and B, respectively. Eighty-four percent and 88% of 2-cell embryos reached the expanded blastocyst stage when cultured in media A and B, respectively. A prospective randomized study was then performed to evaluate this synthetic serum substitute in human in vitro fertilization. Among 141 ovum pick-up (OPU), oocytes retrieved in 74 cases were processed in medium A and oocytes retrieved in 67 others in medium B. In media A and B, the fertilization rate was 67% and 44.3% respectively, and the pregnancy rate/OPU 23% and 9%, respectively. The pregnancy rate/transfer was 28.8% and 12.2% respectively, and the implantation rate/transferred embryo 9.5% and 4.2%. In the human sperm survival assay, the vitality and residual motility after 24 hours of incubation were significantly lower in medium B. In conclusion, UltroSer G successfully sustained the development in vitro of mouse embyros. However in human, it reduced sperm survival, oocyte fertilization, and embryo viability. PMID:2806622

  20. Laser irradiation of mouse spermatozoa enhances in-vitro fertilization and Ca2+ uptake via reactive oxygen species

    NASA Astrophysics Data System (ADS)

    Cohen, Natalie; Lubart, Rachel; Rubinstein, Sara; Breitbart, Haim

    1996-11-01

    630 nm He-Ne laser irradiation was found to have a profound influence on Ca2+ uptake in mouse spermatozoa and the fertilizing potential of these cells. Laser irradiation affected mainly the mitochondrial Ca2+ transport mechanisms. Furthermore, the effect of light was found to be Ca2+-dependent. We demonstrate that reactive oxygen species (ROS) are involved in the cascade of biochemical events evoked by laser irradiation. A causal association between laser irradiation, ROS generation, and sperm function was indicated by studies with ROS scavengers, superoxide dismutase (SOD) and catalase, and exogenous hydrogen peroxide. SOD treatment resulted in increased Ca2+ uptake and in enhanced fertilization rate. Catalase treatment impaired the light-induced stimulation in Ca2+ uptake and fertilization rate. Exogenous hydrogen peroxide was found to enhance Ca2+ uptake in mouse spermatozoa and the fertilizing capability of these cells in a dose-dependent manner. These results suggest that the effect of 630 nm He-Ne laser irradiation is mediated through the generation of hydrogen peroxide by the spermatozoa and that this effect plays a significant role in the augmentation of the sperm cells' capability to fertilize metaphase II-arrested eggs in-vitro.

  1. [Pregnancy and birth of monozygous female twins following in vitro fertilization and embryo transfer].

    PubMed

    Mettler, L; Riedel, H H; Grillo, M; Michelmann, H W; Baukloh, V; Weisner, D; Semm, K; Bastert, G; Hack, H J

    1984-10-01

    The rate of identical twins after in vitro fertilisation and embryo replacement is higher than after physiological conception. The present paper reports on the first delivery of twins (identical twins) after in vitro fertilisation and embryo replacement. The in vitro fertilisation and embryo replacement of one 4-cell stage resulted in a twin pregnancy. The difficult pregnancy, early delivery and intense perinatal care of the children deserves special consideration. In addition the paper gives a survey on the present number of deliveries after in vitro fertilisation and embryo replacement given at the 3rd World Congress on In Vitro Fertilisation and Embryo Transfer at Helsinki in May 1984.

  2. Differential influence of ampullary and isthmic derived epithelial cells on zona pellucida hardening and in vitro fertilization in ovine.

    PubMed

    Dadashpour Davachi, Navid; Zare Shahneh, Ahmad; Kohram, Hamid; Zhandi, Mahdi; Shamsi, Helia; Hajiyavand, Amir M; Saadat, Mozafar

    2016-03-01

    The central role of the oviduct, as the site of zona pellucida (ZP) maturation, fertilization and early embryogenesis, has been recognized. The objective of this study was to investigate whether ampullary and isthmic derived epithelial cells have different effects on in vitro ZP hardening, in vitro fertilization (IVF) and in vitro culture (IVC) of the resulting embryos. Cumulus oocyte complexes (COCs) were matured in a coculture system with ampullary/isthmic epithelial cells, TCM199 supplemented with insulin-like growth factor I (IGF-I) and epithelial derived growth factor (EGF) (GF treated group), conditioned media produced using ampullary (ACM), isthmic (ICM), COCs+ampullary, and COCs+isthmic epithelial cells, contactless culture system, oviductal fluid, GF+ACM/ICM, and drops of TCM199 (control), for 24h. The matured oocytes were randomly divided into two groups: Group I was subjected to ZP digestion; Group II underwent IVF. The duration of the ZP digestion, in a coculture system with ampullary epithelial cells (AE) was significantly increased (p<0.05), compared with other groups. Penetrated oocytes and monospermic fertilization were significantly increased (p<0.05) in the AE group. The mean number of spermatozoa per penetrated oocyte was reduced dramatically for the AE group (p<0.05). A significant increase (p<0.05) in the embryo development was observed in all treated groups, compared to the control. Results revealed that epithelial cells harvested from the ampullary segment of the oviduct had in vitro specialized role in ZP hardening and have subsequent IVF and IVC outcomes.

  3. Does hormonal contraception prior to in vitro fertilization (IVF) negatively affect oocyte yields? - A pilot study

    PubMed Central

    2013-01-01

    Background As oral contraceptives (OCs) suppress anti-Müllerian hormone (AMH), and hormonal contraceptives (HCs), likely, suppress functional ovarian reserve, this study was initiated to determine whether HC affect oocyte yields. Methods We investigated in a retrospective cohort study 43 oocyte donors in 71 in vitro fertilization (IVF) cycles, evaluating anti-Müllerian hormone (AMH) and oocyte yields as reflections of functional ovarian reserve (OR). In 25 IVF cycles egg donors were on HC within one month prior to IVF, and in 46 cycles they were not. Donors, based on their HCs, were further subdivided into 12 with less, and 13 with more androgenic progestins. Results While the three groups did not differ in age, age at menarche, BMI and AMH, oocyte yields among donors who utilized estrane- and gonane-derived (higher androgenic) HCs were lower 11.3 (95% CI 8.3 – 14.3) than either donors using no HCs 16.6 (95% CI 14.7 -18.4) (P < 0.05) or those using anti-androgenic HCs 19.0 (95% CI 12.2-25.8) (P< 0.01). Significance was maintained after adjustments for the donor age and total FSH dose used in ovulation induction. Conclusions Even in young oocyte donors, high androgenic OC exposure appears to suppress functional ovarian reserve and oocyte yields. Since OCs are often routinely used in preparation for IVF, such practice may require reevaluation. Especially in women with diminished ovarian reserve OCs, and especially high androgenic progestin HCs, should, likely, be avoided. PMID:23557032

  4. LOCALIZATION OF THE SPERM PROTEIN SP22 AND INHIBITION OF FERTILITY IN VIVO AND IN VITRO

    EPA Science Inventory

    We previously established that the levels sperm membrane protein SP22 are highly correlated with the fertility of sperm from the cauda epididymidis of rats exposed to both epididymal and testicular toxicants, and that a testis-specific SP22 transcript is expressed in post-meiotic...

  5. Rapamycin Influences the Efficiency of In vitro Fertilization and Development in the Mouse: A Role for Autophagic Activation

    PubMed Central

    Lee, Geun-Kyung; Shin, Hyejin; Lim, Hyunjung Jade

    2016-01-01

    The mammalian target of rapamycin (mTOR) regulates cellular processes such as cell growth, metabolism, transcription, translation, and autophagy. Rapamycin is a selective inhibitor of mTOR, and induces autophagy in various systems. Autophagy contributes to clearance and recycling of macromolecules and organelles in response to stress. We previously reported that vitrified-warmed mouse oocytes show acute increases in autophagy during warming, and suggested that it is a natural response to cold stress. In this follow-up study, we examined whether the modulation of autophagy influences survival, fertilization, and developmental rates of vitrified-warmed mouse oocytes. We used rapamycin to enhance autophagy in metaphase II (MII) oocytes before and after vitrification. The oocytes were then subjected to in vitro fertilization (IVF). The fertilization and developmental rates of vitrified-warmed oocytes after rapamycin treatment were significantly lower than those for control groups. Modulation of autophagy with rapamycin treatment shows that rapamycin-induced autophagy exerts a negative influence on fertilization and development of vitrified-warmed oocytes. PMID:26954158

  6. Serum and follicular fluid concentrations of polybrominated diphenyl ethers and in-vitro fertilization outcome

    PubMed Central

    Johnson, Paula I.; Altshul, Larisa; Cramer, Daniel W.; Missmer, Stacey A.; Hauser, Russ; Meeker, John D.

    2012-01-01

    There is evidence of endocrine disruption and reproductive effects in animals following exposure to certain PBDEs, but human studies are limited. The goal of this study was to investigate the use of serum and follicular fluid as biomarkers of exposure to PBDEs and to explore whether a relationship between PBDE exposure and early pregnancy loss exists. We measured 8 PBDE congeners in archived serum and ovarian follicular fluid samples from 65 women undergoing in-vitro fertilization (IVF). Logistic regression models were used to predict the odds of failed embryo implantation associated with higher levels of PBDEs among the women in the study. There were moderate Kendall’s Tau-beta correlations between serum and follicular fluid concentrations of BDE 28, 47, 100 and 154 (Tβ =0.29–0.38, all p-values<0.005), but BDE 99 and 153 were not correlated between the two matrices (Tβ<0.2, p-values>0.05). Women with detectable concentrations of BDE 153 (39% had detectable levels) in follicular fluid had elevated odds of failed implantation compared with women who had non-detectable concentrations (adjusted OR=10.0; 95%CI: 1.9 to 52; p=0.006; adjusted by age and body mass index). These findings suggest that exposure to BDE 153 may be associated with failed embryo implantation. Due to our observation of only moderate correlations between matrices, serum PBDE concentrations may not be a good indicator of follicular fluid concentrations when studying early pregnancy endpoints in women undergoing IVF. PMID:22572111

  7. Association of FMR1 Genotypes with In Vitro Fertilization (IVF) Outcomes Based on Ethnicity/Race

    PubMed Central

    Gleicher, Norbert; Weghofer, Andrea; Lee, Irene H.; Barad, David H.

    2011-01-01

    The FMR1 gene, mapping to an area of the X chromosome closely associated with autoimmunity also affects ovarian reserve, with specific genotypes associated with distinct ovarian aging patterns. They, therefore, could also be associated with differences of in vitro fertilization (IVF) outcomes, reported between races/ethnicities. We analyzed 339 consecutive IVF patients, 232 Caucasian, 59 African and 48 Asian, for FMR1 genotypes, and tested by multiple logistic regressions for associations between race/ethnicity, FMR1 genotype, autoimmunity and pregnancy chances with IVF. FMR1 genotypes were predictive of pregnancy (P = 0.046), het-norm/low most significantly and with decreasing chance in comparison to norm genotypes (OR 0.44; 95% CI 0.23–0.85; P = 0.014). Race/ethnicity was, overall, independently associated (P = 0.03), African demonstrating decreased odds in comparison to Caucasian (OR 0.33. 95%CI 0.13–0.79; P = 0.014). Autoimmunity did not differ but interaction of autoimmunity with FMR1 genotype almost reached significance (P = 0.07). Logistic regression with race/ethnicity and interaction between FMR1 genotype and autoimmunity in the model, demonstrated 2.5-times the odds of being associated with autoimmune positivity (OR 2.5, 1.34–4.55; P = 0.004). FMR1 genotypes offer a possible explanation for differences in IVF outcomes between races/ethnicities. PMID:21526209

  8. Managing and predicting low response to standard in vitro fertilization therapy: a review of the options.

    PubMed

    Karande, Visjhvanath C

    2003-01-01

    Low responders remain a difficult group of patients to manage in an in vitro fertilization (IVF) program. Such patients have low peak estradiol concentrations (<500 ng/L) and few dominant follicles on the day of human chorionic gonadotropin administration (<5 to <2), and therefore have few retrieved mature oocytes (< or = 4 to < or = 6) with resultant lower pregnancy rates when stimulated with standard IVF therapy (daily gonadotropin dose 150-300IU [2-4 ampoules]). It is difficult to compare the various strategies that have been utilized to manage low responders because the definition of a low responder varies widely. Also, very few large prospective randomized trials have compared different protocols. Two stimulation protocols involving varying doses of leuprorelin and high doses of gonadotropins appear to be the most promising. The early gonadotropin-releasing hormone (GnRH) agonist cessation protocol involves routine pituitary desensitization with luteal phase start of leuprorelin 0.5 mg for 10-14 days. With the onset of gonadotropin treatment, there is cessation of leuprorelin. The 'microdose flare' protocol utilizes oral contraceptive priming followed by diluted doses of leuprorelin 50 microg given twice daily. Two days later, stimulation is started by adding high doses of gonadotropins. The microdoses of leuprorelin and the high doses of gonadotropins are then continued until the day of chorionic gonadotropin administration. Patients who do not respond to these protocols or those with severely diminished ovarian reserve are candidates for donor oocytes. The role of GnRH antagonists and the in vitro maturation of oocytes are potentially exciting strategies that are still being investigated. Ultrasound measurements of ovarian volume, baseline antral follicle counts, and Doppler measurement of ovarian stromal blood flow now make it possible to predict low response to IVF therapy. Low response can be expected if the smaller ovary has a volume <3 cm3, or the mean

  9. Effects of the intensity of downregulation on outcome of in vitro fertilization and embryo transfer.

    PubMed

    Lam, P M; Cheung, L P; Choy, C M Y; Lau, Y P; Haines, C

    2002-04-01

    Gonadotropin releasing hormone agonists (GnRHa) are commonly used during in vitro fertilization and embryo transfer (IVF/ET) treatment cycles to downregulate the hypothalamic-pituitary-ovarian axis prior to ovarian stimulation with gonadotropins. It has been suggested that profound downregulation may have an adverse effect on IVF/ET outcome. The aim of this study was to examine the relationship between the degree of downregulation and IVF/ET outcome. A retrospective analysis was performed on 151 IVF/ET cycles conducted over a six month period. Intensity of downregulation was assessed using measurements of serum concentrations of luteinizing hormone (LH) and estradiol (E2) made at the end of a two week downregulation period. There was no correlation between serum concentration of LH (whether used alone or in combination with E2) and IVF/ET pregnancy rates. However, those subjects who were more suppressed according to the E2 concentration (< 148 pmol/l, [median]) required significantly more gonadotropins (3306 IU versus 2863 IU, p < 0.05) and took longer for follicles to reach maturity (10.9 days versus 9.7 days, p < 0.05). They also had a lower pregnancy rate per embryo transfer (10.4% versus 28.6%, p < 0.05) compared with those having a higher basal E2 concentration. We conclude from this study that the basal serum E2 concentration rather than the LH concentration is a more sensitive indicator of the intensity of downregulation by GnRHa and it may be a better predictor of IVF outcome.

  10. Induction of follicular growth in the squirrel monkey (Saimiri sciureus): enhanced recovery of mature ova for fertilization in vitro.

    PubMed

    Ozasa, H; Gould, K G

    1987-01-01

    In order to improve the rate of recovery of mature ova over those previously reported, we evaluated a new hormonal regimen for induction of follicular growth in the squirrel monkey. This regimen, which consisted of 50 IU pregnant mare's serum (PMS) per day for 4 days followed by 50 IU PMS and 250 IU human chorionic gonadotropin (hCG) on the 5th day, gave an average yield of 2.3 mature ova per animal from 14 experiments involving 11 animals. Ova were recovered through laparotomy or laparoscopy 18 h after PMS and hCG. This number reached 3.3 per animal when ova matured in vitro were included. The fertilization rate was significantly higher for ova matured in vivo (44%) than for in vitro-matured ova (7%) (P less than 0.05), showing the importance of recovering mature ova for successful fertilization in vitro. In agreement with previous observations, a seasonal pattern was noted for the response of animals to induction of follicular development, with a reduction in response occurring during a summer period from July to September. Our observations also suggest the occurrence of immunologic problems associated with the use of PMS in the squirrel monkey.

  11. In vitro fertilization using frozen-thawed feline epididymal spermatozoa from corpus and cauda regions.

    PubMed

    Kunkitti, Panisara; Axnér, Eva; Bergqvist, Ann-Sofi; Sjunnesson, Ylva

    2016-10-01

    Epididymal sperm preservation offers a potential for rescuing genetic material from endangered or valuable animals after injury or death. Spermatozoa from corpus, as well as from cauda, have the capability to be motile and to undergo capacitation and can thus potentially be preserved for assisted reproductive technologies. In the present study, feline frozen-thawed epididymal spermatozoa from corpus and cauda regions were investigated for their ability to fertilize homologous oocytes and further embryo development in vitro. Epididymal spermatozoa from corpus and cauda of seven cats were cryopreserved and used for IVF. Cumulus-oocyte complexes (n = 419) were obtained from female cats after routine spaying. Frozen-thawed corpus epididymal spermatozoa showed similar properties of acrosome integrity, membrane integrity, and chromatin integrity as frozen-thawed spermatozoa from cauda except corpus spermatozoa showed lower motility (P < 0.05). The fertilizing capacity of frozen-thawed corpus epididymal spermatozoa was confirmed by similar number of embryos developing to the two- and four-cell stages compared with sperm from cauda (32.03% vs. 33.33%). However, oocytes fertilized with corpus spermatozoa had lower potential to develop to the blastocyst stage (6.79%) and had lower cell numbers compared to oocytes fertilized with cauda spermatozoa (14.08%). In conclusion, spermatozoa from corpus epididymis had a similar capability to fertilize homologous oocytes in vitro as sperm from cauda but resulted in fewer embryos developing to the blastocyst stage compared to spermatozoa from the cauda. PMID:27242180

  12. The analysis of mitochondrial DNA haplogroups and variants for in vitro fertilization failure in a Han Chinese population.

    PubMed

    Mao, Genhong; Lu, Ping; Huang, Xiao-Hui; Wang, Wu-Liang; Tao, Shi-Bo; Li, Qian; Wang, Xiao-Ling; Wang, Ya-Nan

    2016-07-01

    In this study, we aimed to investigate the associations of mitochondrial DNA (mtDNA) haplogroups and variants with in vitro fertilization (IVF) failure. A retrospective, comparative study of 260 fresh IVF cycles in a Han Chinese population was performed from July 2011 to April 2014. Seventy-three couples had low fertilization rates (≤30%) or total fertilization failure, and 187 controls with normal fertilization were included. Human sperm mtDNA haplogroups and variants were determined by polymerase chain reaction (PCR), nested PCR and direct sequencing. One unreported point variant, A15397G, and two novel deletions at positions 8270-8278 and 8276-8284 were found in this study. A homozygous variant, G9053A in MT-ATP6, was detected in 4 of the 73 cases with fertilization failure, whereas this substitution was not detected in the control group (p < 0.01). The frequency of the point 10397 homozygous variant in MT-ND3 in the IVF failure group was markedly lower than that in the control group (p < 0.05). Furthermore, this study showed that the frequencies of point 8701 and 8943 heterozygous variants in MT-ATP6 in the IVF failure group were also markedly lower than those in the control group (p < 0.05). In addition, the frequency of haplogroup Z was markedly higher in the IVF failure group than in the control group (p < 0.05). Our results suggested that MT-ATP6 variants might be possible causes of IVF failure, but the 10397 homozygous variant in MT-ND3 might help decrease the risk of developing IVF failure. Furthermore, this study indicated that men with haplogroup Z might inherit a higher risk of IVF failure in the Han Chinese population.

  13. Maturity and fertility of rhesus monkey oocytes collected at different intervals after an ovulatory stimulus (human chorionic gonadotropin) in in vitro fertilization cycles.

    PubMed

    Wolf, D P; Alexander, M; Zelinski-Wooten, M; Stouffer, R L

    1996-01-01

    In rhesus monkeys undergoing ovarian stimulation for in vitro fertilization (IVF), a midcycle injection of human chorionic gonadotropin (hCG) substitutes for the LH surge and induces preovulatory oocyte maturation. The time interval between injection and oocyte collection, ideally, allows for the completion of oocyte maturation without ovulation, which would reduce the number of oocytes available for harvest. To evaluate the influence of this time interval on oocyte parameters following hCG administration, we conducted a series of gonadotropin treatment protocols in 51 animals in which the interval from hCG administration to follicular aspiration was systematically varied from 27 to 36 hr. Follicle number and size, evaluated prior to hCG administration by sonography, did not vary significantly or consistently with preovulatory maturation time. Oocytes were harvested by laparotomy or laparoscopy, and scored for maturity before insemination. The percentage of mature, metaphase II (MII) oocytes at recovery increased significantly with increasing preovulatory time and was inversely proportional to that of metaphase I (MI) oocytes. However, oocyte yield tended toward a progressive decrease with increasing preovulatory maturation times from a high of 27 oocytes at 27 hr to a low of 17 oocytes/animal at the 36 hr time interval. Fertilization levels declined significantly from a high of 50% at 27 hr to a low of 30% at 36 hr. Thus, although higher percentages of mature oocytes were recovered at the longer time intervals, optimal oocytes/embryo harvests were realized after the shorter time intervals (27 and 32 hr) and are most compatible with the goal of achieving high yields of fertile oocytes and embryos following gonadotropin stimulation in rhesus monkeys. PMID:8720116

  14. Estrogen supplementation to progesterone as luteal phase support in patients undergoing in vitro fertilization: systematic review and meta-analysis.

    PubMed

    Zhang, Xiao-Mei; Lv, Fang; Wang, Pin; Huang, Xia-Man; Liu, Kai-Feng; Pan, Yu; Dong, Nai-Jun; Ji, Yu-Rong; She, Hong; Hu, Rong

    2015-02-01

    Meta-analyses have found conflicting results with respect to the use of progesterone or progesterone plus estrogen as luteal phase support for in vitro fertilization (IVF) protocols involving gonadotropins and/or gonadotropin-releasing hormone analogs. The aim of the present study was to perform an updated meta-analysis on the efficacy of progesterone versus progesterone plus estrogen as luteal phase support. We searched the MEDLINE, Cochrane Library, and Google Scholar databases (up to March 18, 2014). The search terms were (estrogen OR estradiol OR oestradiol) AND (progesterone) AND (IVF OR in vitro fertilization) AND (randomized OR prospective). We did not limit the form of estrogen and included subjects who contributed more than 1 cycle to a study. The primary outcome was clinical pregnancy rate. Secondary outcomes were ongoing pregnancy rate, fertilization rate, implantation rate, and miscarriage rate. A total of 11 articles were included in the present analysis, with variable numbers of studies assessing each outcome measure. Results of statistical analyses indicated that progesterone plus estrogen treatment was more likely to result in clinical pregnancy than progesterone alone (pooled odds ratio 1.617, 95% confidence interval 1.059-2.471; P = 0.026). No significant difference between the 2 treatment regimens was found for the other outcome measures. Progesterone plus estrogen for luteal phase support is associated with a higher clinical pregnancy rate than progesterone alone in women undergoing IVF, but other outcomes such as ongoing pregnancy rate, fertilization rate, implantation rate, and miscarriage rate are the same for both treatments. PMID:25715250

  15. Estrogen supplementation to progesterone as luteal phase support in patients undergoing in vitro fertilization: systematic review and meta-analysis.

    PubMed

    Zhang, Xiao-Mei; Lv, Fang; Wang, Pin; Huang, Xia-Man; Liu, Kai-Feng; Pan, Yu; Dong, Nai-Jun; Ji, Yu-Rong; She, Hong; Hu, Rong

    2015-02-01

    Meta-analyses have found conflicting results with respect to the use of progesterone or progesterone plus estrogen as luteal phase support for in vitro fertilization (IVF) protocols involving gonadotropins and/or gonadotropin-releasing hormone analogs. The aim of the present study was to perform an updated meta-analysis on the efficacy of progesterone versus progesterone plus estrogen as luteal phase support. We searched the MEDLINE, Cochrane Library, and Google Scholar databases (up to March 18, 2014). The search terms were (estrogen OR estradiol OR oestradiol) AND (progesterone) AND (IVF OR in vitro fertilization) AND (randomized OR prospective). We did not limit the form of estrogen and included subjects who contributed more than 1 cycle to a study. The primary outcome was clinical pregnancy rate. Secondary outcomes were ongoing pregnancy rate, fertilization rate, implantation rate, and miscarriage rate. A total of 11 articles were included in the present analysis, with variable numbers of studies assessing each outcome measure. Results of statistical analyses indicated that progesterone plus estrogen treatment was more likely to result in clinical pregnancy than progesterone alone (pooled odds ratio 1.617, 95% confidence interval 1.059-2.471; P = 0.026). No significant difference between the 2 treatment regimens was found for the other outcome measures. Progesterone plus estrogen for luteal phase support is associated with a higher clinical pregnancy rate than progesterone alone in women undergoing IVF, but other outcomes such as ongoing pregnancy rate, fertilization rate, implantation rate, and miscarriage rate are the same for both treatments.

  16. In vitro fertilization (IVF) in mammals: epigenetic and developmental alterations. Scientific and bioethical implications for IVF in humans.

    PubMed

    Ventura-Juncá, Patricio; Irarrázaval, Isabel; Rolle, Augusto J; Gutiérrez, Juan I; Moreno, Ricardo D; Santos, Manuel J

    2015-12-18

    The advent of in vitro fertilization (IVF) in animals and humans implies an extraordinary change in the environment where the beginning of a new organism takes place. In mammals fertilization occurs in the maternal oviduct, where there are unique conditions for guaranteeing the encounter of the gametes and the first stages of development of the embryo and thus its future. During this period a major epigenetic reprogramming takes place that is crucial for the normal fate of the embryo. This epigenetic reprogramming is very vulnerable to changes in environmental conditions such as the ones implied in IVF, including in vitro culture, nutrition, light, temperature, oxygen tension, embryo-maternal signaling, and the general absence of protection against foreign elements that could affect the stability of this process. The objective of this review is to update the impact of the various conditions inherent in the use of IVF on the epigenetic profile and outcomes of mammalian embryos, including superovulation, IVF technique, embryo culture and manipulation and absence of embryo-maternal signaling. It also covers the possible transgenerational inheritance of the epigenetic alterations associated with assisted reproductive technologies (ART), including its phenotypic consequences as is in the case of the large offspring syndrome (LOS). Finally, the important scientific and bioethical implications of the results found in animals are discussed in terms of the ART in humans.

  17. Downregulation of miR-199a-5p Disrupts the Developmental Potential of In Vitro-Fertilized Mouse Blastocysts.

    PubMed

    Tan, Kun; Wang, Xiaodong; Zhang, Zhenni; Miao, Kai; Yu, Yong; An, Lei; Tian, Jianhui

    2016-09-01

    Although in vitro fertilization (IVF), one of the most effective and successful assisted reproductive technologies, is widely used for treating infertility and in animal breeding, increasing evidence indicates that IVF offspring are linked to various short- or long-term consequences. Erroneous epigenetic modifications induced by IVF are suspected of contributing to these consequences. Among these epigenetic modifications, microRNAs may affect embryo implantation and early postimplantation development. Here, we performed comparative microRNA profiling between in vivo-fertilized (IVO group) and in vitro-fertilized (IVF group) mouse embryos at Embryonic Day 3.5 (E3.5) and E7.5. Our dynamic analyses showed that the dysregulated microRNAs were mainly associated with the regulation of genes involved in carcinogenesis, genetic information processing, glucose metabolism, cytoskeleton organization, and neurogenesis. Further analysis showed that miR-199a-5p was consistently downregulated in IVF embryos compared with their IVO counterparts. Through gain- and loss-of-function experiments, we demonstrated that IVF-induced downregulation of miR-199a-5p results in a higher glycolytic rate and lower developmental potential of IVF blastocysts, including cell lineage misallocation and lower fetal survival post implantation. Therefore, preventing downregulation of miR-199a-5p may become an effective strategy for improving the development of IVF peri-implantation embryos in the future. PMID:27488027

  18. Novel dehydroepiandrosterone troche supplementation improves the serum androgen profile of women undergoing in vitro fertilization

    PubMed Central

    Keane, Kevin N; Hinchliffe, Peter M; Namdar, Navid; Conceicao, Jason L; Newsholme, Philip; Yovich, John L

    2015-01-01

    Dehydroepiandrosterone (DHEA) is the most abundant steroid hormone in the circulation and has potent multifunctional activity. Epidemiological evidence suggests that levels of serum DHEA decrease with advancing age, and this has been associated with onset or progression of various age-related ailments, including cognitive decline and dementia, cardiovascular disease, and obesity. Consequently, these findings have sparked intense research interest in DHEA supplementation as an “antiaging” therapy. Currently, DHEA is being used by 25% of in vitro fertilization (IVF) clinicians as an adjuvant in assisted reproductive programs, yet the therapeutic benefit of DHEA is unclear. Here, we examined the use of novel DHEA-containing oral troches in patients undertaking IVF and investigated the impact of these troches on their serum androgen profile. This retrospective study determined the androgen profile of 31 IVF patients before (baseline) and after DHEA supplementation (with DHEA). Baseline serum measurements of testosterone (total and free), DHEA sulfate (DHEAS), sex hormone-binding globulin (SHBG), and androstenedione were made before and after supplementation. Each patient received DHEA troches containing 25 mg of micronized DHEA, and troches were administered sublingually twice daily for a period of no greater than 4 months. Adjuvant treatment with DHEA boosted the serum concentration of a number of androgen-related analytes, including total and free testosterone, androstenedione, and DHEAS, while serum SHBG remained unchanged. Supplementation also significantly increased the free-androgen index in IVF patients. Interestingly, the increase in serum analyte concentration following DHEA supplementation was found to be dependent on body mass index (BMI), but not individual age. Patients with the lowest BMI (<20.0 kg/m2) tended to have lower testosterone and DHEAS, but higher SHBG and androstenedione levels in comparison with other BMI groups postsupplementation

  19. Novel dehydroepiandrosterone troche supplementation improves the serum androgen profile of women undergoing in vitro fertilization.

    PubMed

    Keane, Kevin N; Hinchliffe, Peter M; Namdar, Navid; Conceicao, Jason L; Newsholme, Philip; Yovich, John L

    2015-01-01

    Dehydroepiandrosterone (DHEA) is the most abundant steroid hormone in the circulation and has potent multifunctional activity. Epidemiological evidence suggests that levels of serum DHEA decrease with advancing age, and this has been associated with onset or progression of various age-related ailments, including cognitive decline and dementia, cardiovascular disease, and obesity. Consequently, these findings have sparked intense research interest in DHEA supplementation as an "antiaging" therapy. Currently, DHEA is being used by 25% of in vitro fertilization (IVF) clinicians as an adjuvant in assisted reproductive programs, yet the therapeutic benefit of DHEA is unclear. Here, we examined the use of novel DHEA-containing oral troches in patients undertaking IVF and investigated the impact of these troches on their serum androgen profile. This retrospective study determined the androgen profile of 31 IVF patients before (baseline) and after DHEA supplementation (with DHEA). Baseline serum measurements of testosterone (total and free), DHEA sulfate (DHEAS), sex hormone-binding globulin (SHBG), and androstenedione were made before and after supplementation. Each patient received DHEA troches containing 25 mg of micronized DHEA, and troches were administered sublingually twice daily for a period of no greater than 4 months. Adjuvant treatment with DHEA boosted the serum concentration of a number of androgen-related analytes, including total and free testosterone, androstenedione, and DHEAS, while serum SHBG remained unchanged. Supplementation also significantly increased the free-androgen index in IVF patients. Interestingly, the increase in serum analyte concentration following DHEA supplementation was found to be dependent on body mass index (BMI), but not individual age. Patients with the lowest BMI (<20.0 kg/m(2)) tended to have lower testosterone and DHEAS, but higher SHBG and androstenedione levels in comparison with other BMI groups postsupplementation. However

  20. Effects of a mindfulness-based intervention on fertility quality of life and pregnancy rates among women subjected to first in vitro fertilization treatment.

    PubMed

    Li, Jing; Long, Ling; Liu, Yu; He, Wei; Li, Min

    2016-02-01

    Generally, undergoing an in vitro fertilization (IVF) treatment is an emotional and physical burden for the infertile woman, which may negatively influence the treatment outcome. We conducted a study to investigate the effectiveness of a mindfulness-based intervention (MBI) among women subjected to first IVF treatment at a fertility medical center in China. Among infertile women registered for their first IVF treatment, 58 completed the intervention, and 50 were assigned to a control group using a non-randomized controlled study. Standardized measures of mindfulness, self-compassion, emotion regulation difficulties, infertility-related coping strategies and fertility quality of life (FertiQoL) were endorsed pre- and post-MBI, and measure of pregnancy rates at the sixth months after the intervention. Both groups were shown to be equivalent at baseline. By the end of the intervention, women who attended the intervention revealed a significant increase in mindfulness, self-compassion, meaning-based coping strategies and all FertiQoL domains. Inversely, they presented a significant decrease in emotion regulation difficulties, active- and passive-avoidance coping strategies. Women in the control group did not present significant changes in any of the psychological measures. Moreover, there were statistically significant differences between participants in the pregnancy rates, the experiment group higher than the control group. Being fully aware of the present moment without the lens of judgment, seems to help women relate to their infertility and IVF treatment in new ways. This is beneficial for promoting their self-compassion, adaptive emotion regulation and infertility-related coping strategies, which, in turn, may influence the FertiQoL and pregnancy rates. The brief and nonpharmaceutical nature of this intervention makes it a promising candidate for women' use during first IVF treatment.

  1. Lessons from elective in vitro fertilization (IVF) in, principally, non-infertile women

    PubMed Central

    2012-01-01

    Background We here report the first investigation of exclusively elective in vitro fertilization (IVF) cycles in women with no apparent history of infertility. Since IVF outcome in women with infertility are always influenced by underlying causes of infertility, a study on non-infertile women may offer new insights. Methods We investigated 88 females without history of infertility in 109 consecutive elective IVF cycles, almost exclusively performed for purposes of preimplantation genetic screening (PGS; i.e., elective gender selection). The following questions were addressed: (i) impact of PGS on IVF pregnancy chances; (ii) impact of transfer of 1 vs. ≥2 embryos on IVF pregnancy chances; (iii) correlation of anti-Müllerian hormone (AMH) levels to embryo ploidy (iv) effect of gonadotropin dosage used in stimulation on available embryos for transfer; and (v) in form of a 1:1 case control study, compared 33 elective PGS cycles with matched control cycles without PGS, performed in couples with either prior tubal ligations and/or severe male factor infertility as indication of IVF. Results The overall clinical pregnancy rate for the group was 36.7%; pregnancy was associated with number of euploid (P = 0.009) and number of embryos transferred (P = 0.001). Odds of pregnancy were 3.4-times higher if ≥4 euploid embryos were produced in comparison to <4 (95% CI 1.2 to 9.2; P = 0.019), and odds of pregnancy were 6.6-times higher if greater than or equal to 2 rather than <1 euploid embryos were transferred (95% CI 2.0 to 21.7; P = 0.002). Increasing AMH (P = 0.001) and gonadotropin dosage used in ovarian stimulation (P = 0.024), was, independently, associated with number of available euploid embryos. Increasing AMH, but not follicle stimulating hormone (FSH), was associated with number of embryos available for biopsy and PGS (P = 0.0001). Implantation rates were 26.4% with PGS and 9.5% without (P = 0.008). Women undergoing PGS, demonstrated

  2. Follicular fluid steroid content and in vitro steroid secretion by granulosa-lutein cells from individual follicles among different stimulation protocols for in vitro fertilization-embryo transfer.

    PubMed

    Hill, G A; Osteen, K G

    1989-08-01

    The in vitro steroidogenic capacity of granulosa-lutein (G-L) cells aspirated from individual follicles during cycles of in vitro fertilization-embryo transfer was examined and compared among three different stimulation protocols: human menopausal gonadotropins (hMG), clomiphene citrate (CC) and hMG, and follicle stimulating hormone (FSH). In addition, the clinical outcome of the patients in each protocol was examined. After 3 days of culture in basal medium, fresh medium with or without androstenedione (A) (10(-7) M) was added for 24 hr, at which time medium was obtained for measurement of progesterone (P) and estradiol (E) content. Follicular fluid (FF) P, E, and A were measured in each follicle and compared among protocols. FF from individual follicles in patients stimulated with FSH contained higher levels of P compared to FF from patients stimulated with hMG or CC/hMG, while E was higher in patients stimulated with CC/hMG compared to FSH or hMG. FF levels of A were not significantly different among the protocols. In vitro steroid secretion revealed a progressive increase in P secretion in contrast to decreasing E secretion when one compares CC/hMG, hMG, and FSH. Patients undergoing ovarian hyperstimulation with FSH had significantly more atretic oocytes identified at the time of oocyte harvest compared to patients undergoing ovarian hyperstimulation with CC/hMG or hMG. The hMG protocol yielded significantly fewer fertilized oocytes, cleaved embryos, and transferred embryos, compared to the CC/hMG and FSH protocol, however, there was no significant difference in pregnancy rate among the three protocols. These data demonstrate that individual follicles contain G-L cells with markedly different abilities to luteinize in vitro as assessed by steroid secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. In Vitro Fertilization Outcomes and Alcohol Consumption in At-Risk Drinkers: The Effects of a Randomized Intervention

    PubMed Central

    Rossi, Brooke V.; Chang, Grace; Berry, Katharine F.; Hornstein, Mark D.; Missmer, Stacey A.

    2013-01-01

    Background and Objectives Women’s use of alcohol in pregnancy is associated with an increase risk of fetal loss and birth defects. Also, alcohol use in women decreases the success of infertility treatment, such as in vitro fertilization (IVF). Our goal was to determine if there were differences in IVF outcomes and alcohol use parameters among at-risk drinkers randomized to a brief intervention (BI) vs. assessment only (AO). Methods We conducted a randomized controlled trial to determine the effect of brief intervention (BI) or assessment only (AO) among at-risk drinkers on in vitro fertilization (IVF). We studied 37 women (AO= 21; BI= 16). Results While the BI group had a significantly greater decrease in the number of drinks/drinking day compared to the AO group (P=0.04), there were no differences in the likelihood of implantation failure, chemical pregnancy, spontaneous abortion, preterm birth, or live birth. Conclusions BI and AO contributed to a decrease in alcohol use and did not demonstrate differences in IVF outcomes. A larger study may confirm these preliminary findings. Scientific Significance Our results will assist care providers in treating alcohol use in pregnancy in an effective way, such that IVF cycles and the chance of pregnancy are optimized. PMID:23952894

  4. A new rolling culture-based in vitro fertilization system capable of reducing polyspermy in porcine oocytes.

    PubMed

    Kitaji, Hideki; Ookutsu, Shoji; Sato, Masahiro; Miyoshi, Kazuchika

    2015-05-01

    The high incidence of polyspermy is one of the major obstacles during in vitro fertilization (IVF) in pigs. To overcome this, we developed a novel IVF method, which involves constant rotation. Oocytes matured in vitro were mixed with spermatozoa (0.2 × 10(5) sperm/mL) in an IVF medium (200 μL) using a 200 μL PCR tube. This tube was then rotated at 1 rpm for 6 h at 38.5°C in a rotation mixer (experimental group). A second PCR tube was simultaneously cultured without rotation (control group). The rate of polyspermy was evaluated 12 h after insemination and was significantly (P < 0.05; 21.0% vs. 48.3%) lower in the experimental group than in the control group. Sperm penetration rate was similar in oocytes from the experimental and control groups (75.2% vs. 83.1%). However, monospermic fertilization rate of the oocytes was significantly (P < 0.05; 44.8% vs. 21.2%) higher in the experimental group than in the control group. Furthermore, the rate of blastocyst formation (30.1% vs. 20.8%) increased in the experimental group, as compared to the control group. This present system will contribute to increase the efficacy of blastocyst production through reduction of polyspermic penetration.

  5. In vitro colchicine-induced polyploid plantlet production and regeneration from leaf explants of the diploid pear (Pyrus communis L.) cultivar, 'Fertility'

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyploid plantlets, including triploid, tetraploid and other polyploids, were induced from in vitro leaves of a European pear (Pyrus communis L.) cultivar 'Fertility' by a colchicine treatment. In vitro leaves were incubated in 0.4% colchicine solution for 24, 48 or 72 h, and transferred to an adv...

  6. GnRH agonist versus GnRH antagonist in in vitro fertilization and embryo transfer (IVF/ET).

    PubMed

    Depalo, Raffaella; Jayakrishan, K; Garruti, Gabriella; Totaro, Ilaria; Panzarino, Mariantonietta; Giorgino, Francesco; Selvaggi, Luigi E

    2012-04-13

    Several protocols are actually available for in Vitro Fertilization and Embryo Transfer. The review summarizes the main differences and the clinic characteristics of the protocols in use with GnRH agonists and GnRH antagonists by emphasizing the major outcomes and hormonal changes associated with each protocol. The majority of randomized clinical trials clearly shows that in "in Vitro" Fertilization and Embryo Transfer, the combination of exogenous Gonadotropin plus a Gonadotropin Releasing Hormone (GnRH) agonist, which is able to suppress pituitary FSH and LH secretion, is associated with increased pregnancy rate as compared with the use of gonadotropins without a GnRH agonist. Protocols with GnRH antagonists are effective in preventing a premature rise of LH and induce a shorter and more cost-effective ovarian stimulation compared to the long agonist protocol. However, a different synchronization of follicular recruitment and growth occurs with GnRH agonists than with GnRH antagonists. Future developments have to be focused on timing of the administration of GnRH antagonists, by giving a great attention to new strategies of stimulation in patients in which radio-chemotherapy cycles are needed.

  7. Multiple pregnancy after in-vitro fertilization and embryo transfer: report of a quadruplet pregnancy and delivery.

    PubMed

    Goldman, J A; Feldberg, D; Ashkenazi, J; Shelef, M; Dicker, D; Hart, J

    1987-08-01

    The first baby from in-vitro fertilization (IVF) was born in England in 1978 as a result of retrieval of a single preovulatory oocyte in the course of a natural cycle (Steptoe and Edwards, 1978). At present most programmes of IVF throughout the world do not use natural cycles producing only one oocyte, but rather multiple oocyte cycles produced by clomiphene citrate (CC), human menopausal gonadotrophin (HMG), or pure follicle stimulating hormone (FSH), either separately or in combination, sequentially or concomitantly, for the induction of multiple follicular maturation.

  8. The Effect of Extremely Low Frequency Pulsed Electromagnetic Field on In Vitro Fertilization Success Rate in N MRI Mice

    PubMed Central

    Hafizi, Leili; Sazgarnia, Ameneh; Mousavifar, Nezhat; Karimi, Mohammad; Ghorbani, Saleh; Kazemi, Mohammad Reza; Emami Meibodi, Neda; Hosseini, Golkoo; Mostafavi Toroghi, Hesam

    2014-01-01

    Objective: The effects of exposure to electromagnetic fields (EMF) on reproduction systems have been widely debated. In this study, we aimed to investigate whether low frequency EMF could ameliorate the in vitro fertilization success rate in Naval medical research institute (NMRI) Mice. Materials and Methods: In this randomized comparative animal study, ten NMRI mice were randomly divided into 2 equal groups (control and experimental). 10 IU of human chorionic gonadotropin (hCG) was injected intraperitoneally to both groups in order to stimulate ovulating, and ovums were then aspirated and kept in KSOM (modified version of sequential simplex optimization medium with a higher K+ concentration) culture medium. Metaphase II ovums were separated, and sperms obtained by "swim out" method were added to metaphase II ovums in the culture medium. The experimental group was exposed to 1.3 millitesla pulsed electromagnetic field at 4 kilohertz frequency for 5 hours. To assess the efficacy, we considered the identification of two-pronuclear zygote (2PN) under microscope as fertilizing criterion. Results: Total number of collected ovums in the control and experimental groups was 191 and 173, respectively, from which 58 (30.05%) and 52 (30.36%) ovums were collected from metaphase II, respectively. In vitro fertilization (IVF) success rate was 77% in extremely low frequency- pulsed electromagnetic field (ELFPEMF) for exposed group (experimental), whereas the rate was 68% for control group. Conclusion: Despite increased percentile of IVF success rate in exposed group, there was no statistically significant difference between 2 groups, but this hypothesis has still been stated as a question. Further studies with larger sample sizes and different EMF designs are suggested. PMID:24381855

  9. A Case of Secondary Abdominal Pregnancy after in Vitro Fertilization Pre-Embryo Transfer (IVF-ET)

    PubMed Central

    Angelova, Mariya Angelova; Kovachev, Emil Georgiev; Kozovski, Ivan; Kornovski, Yavor Dimitrov; Kisyov, Stefan Vasilev; Ivanova, Vilislava Robert

    2015-01-01

    The authors describe a rare case of secondary abdominal pregnancy after in vitro fertilization pre-embryo transfer (IVF-ET). Ultrasonography was applied to image ectopic gestational sac containing a yolk vesicle and located adjacent to the anterior uterine wall and left adnexa. Laparoscopy was done on the same day followed by sinistral salpingectomy due to tubal abortion indications. Intraabdominal examination showed chorionic structures penetrating pl. vesicouterina. Histological tests confirmed the EP diagnosis in the second material, i.e. indications of secondary abdominal pregnancy. PMID:27275263

  10. Effect of embryo density on in vitro development and gene expression in bovine in vitro-fertilized embryos cultured in a microwell system.

    PubMed

    Sugimura, Satoshi; Akai, Tomonori; Hashiyada, Yutaka; Aikawa, Yoshio; Ohtake, Masaki; Matsuda, Hideo; Kobayashi, Shuji; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei

    2013-01-01

    To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 μl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density. PMID:23154384

  11. Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System

    PubMed Central

    SUGIMURA, Satoshi; AKAI, Tomonori; HASHIYADA, Yutaka; AIKAWA, Yoshio; OHTAKE, Masaki; MATSUDA, Hideo; KOBAYASHI, Shuji; KOBAYASHI, Eiji; KONISHI, Kazuyuki; IMAI, Kei

    2012-01-01

    Abstract To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 µl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density. PMID:23154384

  12. Effect of gibberellic acid on the quality of sperm and in vitro fertilization outcome in adult male rats.

    PubMed

    Hosseinchi, Mohammadreza; Soltanalinejad, Farhad; Najafi, Gholamreza; Roshangar, Leila

    2013-01-01

    Gibberellic acid (GA3) is a group of plant hormones identified in various plants. The aim of this study was to determine the effects of GA3 on sperm parameters and in vitro fertilization (IVF). Fifty six adult male rats were divided into seven groups as, control, treatment and sham. Following 15, 30 and 45 days of GA3 and methanol alcohol (MA) administration, rats were euthanized and epididymis tail was transferred to human tubular fluid (HTF) medium containing 4 mg mL(-1) bovine serum albumin (BSA) .Total number of sperms, the percentage of live sperms, immature sperms and sperms with damaged chromatin and IVF were examined. The oocytes were obtained from immature rats after the injection of pregnant mare's serum (PMSG) and human chorionic gonadotropin (HCG) hormones. Human tubular fluid was used as the fertilization medium and zygotes transferred to fresh 1-cell rat embryos culture medium (mR1ECM) to reach the blastocyst stage. This study showed that GA3 could decrease the number of total sperms on days 30 and 45 in treated group comparison with the control and sham groups. Additionally, GA3 increased the immature sperms and sperms with damaged chromatin. The percentage of fertilization, two-cell embryos and blastocyst resulting from the treatment group on days 30 and 45 also decreased and showed significant differences with the control and sham groups (p < 0.05). The results obtained from this study indicated that the oral use of GA3 could reduce the fertility in rats by influencing the sperm number and the quality of sperm's chromatins. PMID:25568681

  13. Hollow fiber vitrification provides a novel method for cryopreserving in vitro maturation/fertilization-derived porcine embryos.

    PubMed

    Maehara, Miki; Matsunari, Hitomi; Honda, Kasumi; Nakano, Kazuaki; Takeuchi, Yasuhiro; Kanai, Takahiro; Matsuda, Taisuke; Matsumura, Yukina; Hagiwara, Yui; Sasayama, Norihisa; Shirasu, Akio; Takahashi, Masashi; Watanabe, Masahito; Umeyama, Kazuhiro; Hanazono, Yutaka; Nagashima, Hiroshi

    2012-06-01

    In vitro matured (IVM) oocytes have been used to create genetically modified pigs for various biomedical purposes. However, porcine embryos derived from IVM oocytes are very cryosensitive. Developing improved cryopreservation methods would facilitate the production of genetically modified pigs and also accelerate the conservation of genetic resources. We recently developed a novel hollow fiber vitrification (HFV) method; the present study was initiated to determine whether this new method permits the cryopreservation of IVM oocyte-derived porcine embryos. Embryos were created from the in vitro fertilization of IVM oocytes with frozen-thawed sperm derived from a transgenic pig carrying a humanized Kusabira-Orange (huKO) gene. Morula-stage embryos were assigned to vitrification and nonvitrification groups to compare their in vitro and in vivo developmental abilities. Vitrified morulae developed to the blastocyst stage at a rate similar to that of nonvitrified embryos (66/85, 77.6% vs. 67/84, 79.8%). Eighty-eight blastocysts that developed from vitrified morulae were transferred into the uteri of three recipient gilts. All three became pregnant and produced a total of 17 piglets (19.3%). This piglet production was slightly lower, albeit not significantly, than that of the nonvitrification group (27/88, 30.7%). Approximately half of the piglets in the vitrification (10/17, 58.8%) and nonvitrification (15/27, 55.6%) groups were transgenic. There was no significant difference in the growth rates among the piglets in the two groups. These results indicate that the HFV method is an extremely effective method for preserving cryosensitive embryos such as porcine in vitro maturation/fertilization-derived morulae.

  14. Heterotopic Pregnancy After In Vitro Fertilization and Embryo Transfer After Bilateral Total Salpingectomy/Tubal Ligation: Case Report and Literature Review.

    PubMed

    Xu, Ying; Lu, Yingli; Chen, Huiling; Li, Dandan; Zhang, Jingwen; Zheng, Lianwen

    2016-01-01

    Heterotopic pregnancy is defined as the simultaneous occurrence of intrauterine and ectopic pregnancy, either of which may be single or multiple. It occurs in up to 1% of pregnancies after in vitro fertilization and embryo transfer. This article reports 2 rare cases of heterotopic pregnancy after in vitro fertilization and presents a literature review. In the first case, a 28-year-old woman had previous laparoscopic bilateral total salpingectomy for a right tubal pregnancy and a left hydrosalpinx. However, she had ovarian heterotopic pregnancy after a third in vitro fertilization cycle. Emergency laparotomy was performed. The synchronous intrauterine pregnancy continued with no further complications and ended in the delivery of a singleton term pregnancy. The second case combined interstitial and intrauterine pregnancies after bilateral tubal ligation for hydrosalpinges followed by in vitro fertilization and frozen embryo transfer. The possibility of heterotopic pregnancy after bilateral total salpingectomy/tubal ligation, although extremely rare, should also be considered by gynecologists when they treat an in vitro fertilization patient even though an intrauterine pregnancy has been confirmed.

  15. Developmental failure of hybrid embryos generated by in vitro fertilization of water buffalo (Bubalus bubalis) oocyte with bovine spermatozoa.

    PubMed

    Patil, Shekhar; Totey, Satish

    2003-03-01

    The developmental potential of inter-species hybrid embryos produced by in vitro fertilization of in vitro matured buffalo oocytes with bovine spermatozoa was studied with a view to investigate pre-implantation embryo development and its gross morphology, early embryonic gene expression, and embryonic genome activation. Fertilization events with both buffalo and cattle spermatozoa were almost similar. Overall fertilization rate with cattle spermatozoa was 78.4% was not significantly different from that of buffalo spermatozoa (80.2%). Initial cleavage rate between buffalo and hybrid embryo was also similar, and there was no significant difference in their developmental rate till 8-cell stage (26.0 +/- 4.1 vs. 24.3 +/- 4.8). However, only 5.3% of hybrid embryos developed into blastocyst stage compared to 21.7% in buffalo. mRNA phenotyping of insulin-like growth factor family (Insulin, insulin receptor, IGF-I, IGF-I receptor, IGF-II, and IGF-II receptor) and glucose transporter isoforms (GLUT-I, II, III, IV) in hybrid embryos clearly showed that these molecules were not expressed after 8-cell stage onward. Similarly, as observed in buffalo embryos, incorporation of (35)S-methionine and (3)H-uridine could not be observed in hybrid embryos from 8-cell stage onward. This suggests that the maternal-zygotic genome activation did not occur in hybrid embryos. Differential staining also showed that the blastomere stopped dividing after 8-cell stage. Collectively, these parameters clearly showed that there was developmental failure of hybrid embryos.

  16. An Overview of The Available Methods for Morphological Scoring of Pre-Implantation Embryos in In Vitro Fertilization

    PubMed Central

    Nasiri, Nahid; Eftekhari-Yazdi, Poopak

    2015-01-01

    Assessment of embryo quality in order to choose the embryos that most likely result in pregnancy is the critical goal in assisted reproductive technologies (ART). The current trend in human in vitro fertilization/embryo transfer (IVF/ET) protocols is to decrease the rate of multiple pregnancies after multiple embryo transfer with maintaining the pregnancy rate at admissible levels (according to laboratory standards). Assessment of morphological feathers as a reliable non-invasive method that provides valuable information in prediction of IVF/intra cytoplasmic sperm injection (ICSI) outcome has been frequently proposed in recent years. This article describes the current status of morphological embryo evaluation at different pre-implantation stages. PMID:25685730

  17. Analysis of in vivo oocyte maturation, in vitro embryo development and gene expression in cumulus cells of dairy cows and heifers selected for one fertility quantitative trait loci (QTL) located on BTA3.

    PubMed

    Coyral-Castel, S; Brisard, D; Touzé, J-L; Dupont, M; Ramé, C; Uzbekova, S; Dupont, J

    2012-06-01

    We have previously shown that Holstein cows selected for their homozygous favorable ("fertil+") or unfavorable ("fertil-") haplotype at one quantitative trait loci (QTL) of female fertility located on chromosome 3 (QTL-F-Fert-BTA3) had a different success rate 35 and 90 days after the first artificial insemination. To determine whether the lower fertility in "fertil-" animals could be related to oocyte quality, we analyzed the embryo development rate in vitro and the oocyte meiotic maturation in vivo in "fertil+" and "fertil-" heifers. In vitro maturation and fertilization of immature oocytes recovered by ovum pick-up from "fertil+" and "fertil-" heifers resulted in similar cleavage and blastocyst rates in the two haplotypes. However the percentage of expanded blastocysts and the number of cells per blastocyst were significantly higher in "fertil+". Oocytes from presumptive preovulatory follicles were analyzed after ovarian stimulation. A similar rate of immature (from prophase to metaphase-I) and mature oocytes (metaphase-II) was obtained in the two haplotypes, whereas a significantly higher percentage of oocytes from metaphase-I to metaphase-II was observed in "fertil+" compared to "fertil-" heifers. Since cumulus cells (CCs) could reflect the developmental competence of oocytes, we analyzed the expression of seven genes included in the QTL-F-Fert-BTA3 using real-time PCR in bovine CCs after in vivo or in vitro maturation, as a model of higher and lower competence, respectively. Transcript levels of TAGLN2, EEF1A1 and PIGM were higher in CCs after in vitro maturation (IVM) compared to in vivo maturation, whereas no difference was observed for IFI16, KIRREL, SPTA1 and PEX19 expression. The expression levels of all these genes in in preovulatory CCs were not significantly different between "fertil+" and "fertil-" heifers. In conclusion, the lower fertility of "fertil-" females could be partially due to a lowest quality of the oocytes and consequently of

  18. Decreased in vitro fertility in male rats exposed to fluoride-induced oxidative stress damage and mitochondrial transmembrane potential loss

    SciTech Connect

    Izquierdo-Vega, Jeannett A.; Sanchez-Gutierrez, Manuel; Razo, Luz Maria del

    2008-08-01

    Fluorosis, caused by drinking water contamination with inorganic fluoride, is a public health problem in many areas around the world. The aim of the study was to evaluate the effect of environmentally relevant doses of fluoride on in vitro fertilization (IVF) capacity of spermatozoa, and its relationship to spermatozoa mitochondrial transmembrane potential ({delta}{psi}{sub m}). Male Wistar rats were administered at 5 mg fluoride/kg body mass/24 h, or deionized water orally for 8 weeks. We evaluated several spermatozoa parameters in treated and untreated rats: i) standard quality analysis, ii) superoxide dismutase (SOD) activity, iii) the generation of superoxide anion (O{sub 2}{sup {center_dot}}{sup -}), iv) lipid peroxidation concentration, v) ultrastructural analyses of spermatozoa using transmission electron microscopy, vi) {delta}{psi}{sub m}, vii) acrosome reaction, and viii) IVF capability. Spermatozoa from fluoride-treated rats exhibited a significant decrease in SOD activity ({approx} 33%), accompanied with a significant increase in the generation of O{sub 2}{sup {center_dot}} ({approx} 40%), a significant decrease in {delta}{psi}{sub m} ({approx} 33%), and a significant increase in lipid peroxidation concentration ({approx} 50%), relative to spermatozoa from the control group. Consistent with this finding, spermatozoa from fluoride-treated rats exhibited altered plasmatic membrane. In addition, the percentage of fluoride-treated spermatozoa capable of undergoing the acrosome reaction was decreased relative to control spermatozoa (34 vs. 55%), while the percentage fluoride-treated spermatozoa capable of oocyte fertilization was also significantly lower than the control group (13 vs. 71%). These observations suggest that subchronic exposure to fluoride causes oxidative stress damage and loss of mitochondrial transmembrane potential, resulting in reduced fertility.

  19. Laser-assisted in vitro fertilization facilitates fertilization of vitrified-warmed C57BL/6 mouse oocytes with fresh and frozen-thawed spermatozoa, producing live pups.

    PubMed

    Woods, Stephanie E; Qi, Peimin; Rosalia, Elizabeth; Chavarria, Tony; Discua, Allan; Mkandawire, John; Fox, James G; García, Alexis

    2014-01-01

    The utility of cryopreserved mouse gametes for reproduction of transgenic mice depends on development of assisted reproductive technologies, including vitrification of unfertilized mouse oocytes. Due to hardening of the zona pellucida, spermatozoa are often unable to penetrate vitrified-warmed (V-W) oocytes. Laser-assisted in vitro fertilization (LAIVF) facilitates fertilization by allowing easier penetration of spermatozoa through a perforation in the zona. We investigated the efficiency of V-W C57BL/6NTac oocytes drilled by the XYClone laser, compared to fresh oocytes. By using DAP213 for cryoprotection, 83% (1,470/1,762) of vitrified oocytes were recovered after warming and 78% were viable. Four groups were evaluated for two-cell embryo and live offspring efficiency: 1) LAIVF using V-W oocytes, 2) LAIVF using fresh oocytes, 3) conventional IVF using V-W oocytes and 4) conventional IVF using fresh oocytes. First, the groups were tested using fresh C57BL/6NTac spermatozoa (74% motile, 15 million/ml). LAIVF markedly improved the two-cell embryo efficiency using both V-W (76%, 229/298) and fresh oocytes (69%, 135/197), compared to conventional IVF (7%, 12/182; 6%, 14/235, respectively). Then, frozen-thawed C57BL/6NTac spermatozoa (35% motile, 15 million/ml) were used and LAIVF was again found to enhance fertilization efficiency, with two-cell embryo rates of 87% (298/343) using V-W oocytes (P<0.05, compared to fresh spermatozoa), and 73% (195/266) using fresh oocytes. Conventional IVF with frozen-thawed spermatozoa using V-W (6%, 10/168) and fresh (5%, 15/323) oocytes produced few two-cell embryos. Although live offspring efficiency following embryo transfer was greater with conventional IVF (35%, 18/51; LAIVF: 6%, 50/784), advantage was seen with LAIVF in live offspring obtained from total oocytes (5%, 50/1,010; conventional IVF: 2%, 18/908). Our results demonstrated that zona-drilled V-W mouse oocytes can be used for IVF procedures using both fresh and frozen

  20. Laser-Assisted In Vitro Fertilization Facilitates Fertilization of Vitrified-Warmed C57BL/6 Mouse Oocytes with Fresh and Frozen-Thawed Spermatozoa, Producing Live Pups

    PubMed Central

    Woods, Stephanie E.; Qi, Peimin; Rosalia, Elizabeth; Chavarria, Tony; Discua, Allan; Mkandawire, John; Fox, James G.; García, Alexis

    2014-01-01

    The utility of cryopreserved mouse gametes for reproduction of transgenic mice depends on development of assisted reproductive technologies, including vitrification of unfertilized mouse oocytes. Due to hardening of the zona pellucida, spermatozoa are often unable to penetrate vitrified-warmed (V-W) oocytes. Laser-assisted in vitro fertilization (LAIVF) facilitates fertilization by allowing easier penetration of spermatozoa through a perforation in the zona. We investigated the efficiency of V-W C57BL/6NTac oocytes drilled by the XYClone laser, compared to fresh oocytes. By using DAP213 for cryoprotection, 83% (1,470/1,762) of vitrified oocytes were recovered after warming and 78% were viable. Four groups were evaluated for two-cell embryo and live offspring efficiency: 1) LAIVF using V-W oocytes, 2) LAIVF using fresh oocytes, 3) conventional IVF using V-W oocytes and 4) conventional IVF using fresh oocytes. First, the groups were tested using fresh C57BL/6NTac spermatozoa (74% motile, 15 million/ml). LAIVF markedly improved the two-cell embryo efficiency using both V-W (76%, 229/298) and fresh oocytes (69%, 135/197), compared to conventional IVF (7%, 12/182; 6%, 14/235, respectively). Then, frozen-thawed C57BL/6NTac spermatozoa (35% motile, 15 million/ml) were used and LAIVF was again found to enhance fertilization efficiency, with two-cell embryo rates of 87% (298/343) using V-W oocytes (P<0.05, compared to fresh spermatozoa), and 73% (195/266) using fresh oocytes. Conventional IVF with frozen-thawed spermatozoa using V-W (6%, 10/168) and fresh (5%, 15/323) oocytes produced few two-cell embryos. Although live offspring efficiency following embryo transfer was greater with conventional IVF (35%, 18/51; LAIVF: 6%, 50/784), advantage was seen with LAIVF in live offspring obtained from total oocytes (5%, 50/1,010; conventional IVF: 2%, 18/908). Our results demonstrated that zona-drilled V-W mouse oocytes can be used for IVF procedures using both fresh and frozen

  1. Improved in vitro fertilization and development by use of modified human tubal fluid and applicability of pronucleate embryos for cryopreservation by rapid freezing in inbred mice.

    PubMed

    Kito, Seiji; Hayao, Tatsuo; Noguchi-Kawasaki, Yoshiko; Ohta, Yuki; Hideki, Uhara; Tateno, Shintaro

    2004-10-01

    We examined in vitro fertilizability and development of 10 inbred mouse strains (C57BL/6J, C57BL/10, C57BL/10.D2/newSn, C57BL/10-Thy1.1, C57BL/10.Br/Sn, C3H/He, RFM/Ms, STS/A, BALB/c-nu and C.B-17/Icr), and the viability of frozen-thawed in vitro fertilized (IVF) embryos after embryo transfer (ET). In seven strains, fertilizability was significantly greater in modified human tubal fluid (mHTF) compared with modified Krebs-Ringer's bicarbonate solution (TYH medium). The TYH medium supported almost no fertilization in four strains. More than 80% of IVF embryos developed to the blastocyst stage by 120 h in potassium-enhanced simplex optimization medium (KSOM). Reciprocal fertilization between C57BL/6J and BALB/c-nu gametes in TYH medium yielded poor fertilization o f BALB/c-nu due to spermatozoal deficiencies. Increased concentrations of bovine serum albumin and spermatozoa during capacitation and Percoll washing did not drastically affect fertilization. The mHTF, but not TYH medium, supported BALB/c-nu spermatozoa penetration into the zona pellucida irrespective of capacitation media. In vitro fertilized embryos frozen-thawed rapidly were transferred to surrogate mothers at the two-cell stage. Compared with that of unfrozen controls, rapid freezing had no significant effect on fetus development except in C57BL/10.D2/newSn mice. These results suggest that mHTF medium is superior with respect to IVF of inbred mice, and that KSOM adequately supports in vitro fertilized embryo development in inbred mice. The data also indicate that rapid freezing of pronucleate embryos following IVF is suitable for cryopreservation and embryo banking of inbred mice and for the production of genetically modified mice. PMID:15575371

  2. Embryological outcomes in cycles with human oocytes containing large tubular smooth endoplasmic reticulum clusters after conventional in vitro fertilization.

    PubMed

    Itoi, Fumiaki; Asano, Yukiko; Shimizu, Masashi; Honnma, Hiroyuki; Murata, Yasutaka

    2016-01-01

    There have been no studies analyzing the effect of large aggregates of tubular smooth endoplasmic reticulum (aSERT) after conventional in vitro fertilization (cIVF). The aim of this study was to investigate whether aSERT can be identified after cIVF and the association between the embryological outcomes of oocytes in cycles with aSERT. This is a retrospective study examining embryological data from cIVF cycles showing the presence of aSERT in oocytes 5-6 h after cIVF. To evaluate embryo quality, cIVF cycles with at least one aSERT-metaphase II (MII) oocyte observed (cycles with aSERT) were compared to cycles with normal-MII oocytes (control cycles). Among the 4098 MII oocytes observed in 579 cycles, aSERT was detected in 100 MII oocytes in 51 cycles (8.8%). The fertilization rate, the rate of embryo development on day 3 and day 5-6 did not significantly differ between cycles with aSERT and control group. However, aSERT-MII oocytes had lower rates for both blastocysts and good quality blastocysts (p < 0.05). aSERT can be detected in the cytoplasm by removing the cumulus cell 5 h after cIVF. However, aSERT-MII oocytes do not affect other normal-MII oocytes in cycles with aSERT.

  3. GnRH agonist versus GnRH antagonist in in vitro fertilization and embryo transfer (IVF/ET)

    PubMed Central

    2012-01-01

    Several protocols are actually available for in Vitro Fertilization and Embryo Transfer. The review summarizes the main differences and the clinic characteristics of the protocols in use with GnRH agonists and GnRH antagonists by emphasizing the major outcomes and hormonal changes associated with each protocol. The majority of randomized clinical trials clearly shows that inin Vitro” Fertilization and Embryo Transfer, the combination of exogenous Gonadotropin plus a Gonadotropin Releasing Hormone (GnRH) agonist, which is able to suppress pituitary FSH and LH secretion, is associated with increased pregnancy rate as compared with the use of gonadotropins without a GnRH agonist. Protocols with GnRH antagonists are effective in preventing a premature rise of LH and induce a shorter and more cost-effective ovarian stimulation compared to the long agonist protocol. However, a different synchronization of follicular recruitment and growth occurs with GnRH agonists than with GnRH antagonists. Future developments have to be focused on timing of the administration of GnRH antagonists, by giving a great attention to new strategies of stimulation in patients in which radio-chemotherapy cycles are needed. PMID:22500852

  4. Pregnancy, Delivery, and Neonatal Outcomes of In Vitro Fertilization-Embryo Transfer in Patient with Previous Cesarean Scar

    PubMed Central

    Zhang, Ningyuan; Chen, Hua; Xu, Zhipeng; Wang, Bin; Sun, Haixiang; Hu, Yali

    2016-01-01

    Background What role should previous cesarean section play in affecting clinical pregnancy outcomes and avoiding the complications of in vitro fertilization? In this article, we focus on elective single-embryo transfer (eSET) versus double-embryo transfer (DET) and assess the clinical efficacy and safety of eSET in patients who have a previous cesarean scar. Material/Methods The pregnancy, delivery, and neonatal outcomes of 130 patients who had a previous cesarean scar and received in vitro fertilization-embryo transfer (IVF-ET) were retrospectively analyzed. The number of transferred embryos was chosen depending on patients’ desire after acknowledging all benefits and risks, including eSET (eSET group, n=56) and DET (DET group, n=74). A total of 101 patients with previous vaginal delivery receiving IVF-ET in the same period were included as a control group. Results The pregnancy rates, multiple birth rates, abortion rates, ectopic pregnancy rates, gestational age at delivery, preterm birth rates, neonatal birth weight, and take-home baby rates were similar between the previous cesarean section group and the previous vaginal delivery group. A previous cesarean section scar did not affect embryo implantation and pregnancy outcomes in IVF. In the eSET and DET groups of previous cesarean section patients, the embryo implantation rates, pregnancy rates, abortion rates, and take-home baby rates were similar. However, the rate of multiple pregnancies reached 50% in the DET group, which led to more preterm births and lower birth weight. Conclusions Elective single-embryo transfer is a well-accepted strategy to avoid multiple pregnancies and improve the obstetric and neonatal outcomes of singleton pregnancy in IVF patients with a previous cesarean section. PMID:27636504

  5. Pregnancy, Delivery, and Neonatal Outcomes of In Vitro Fertilization-Embryo Transfer in Patient with Previous Cesarean Scar.

    PubMed

    Zhang, Ningyuan; Chen, Hua; Xu, Zhipeng; Wang, Bin; Sun, Haixiang; Hu, Yali

    2016-01-01

    BACKGROUND What role should previous cesarean section play in affecting clinical pregnancy outcomes and avoiding the complications of in vitro fertilization? In this article, we focus on elective single-embryo transfer (eSET) versus double-embryo transfer (DET) and assess the clinical efficacy and safety of eSET in patients who have a previous cesarean scar. MATERIAL AND METHODS The pregnancy, delivery, and neonatal outcomes of 130 patients who had a previous cesarean scar and received in vitro fertilization-embryo transfer (IVF-ET) were retrospectively analyzed. The number of transferred embryos was chosen depending on patients' desire after acknowledging all benefits and risks, including eSET (eSET group, n=56) and DET (DET group, n=74). A total of 101 patients with previous vaginal delivery receiving IVF-ET in the same period were included as a control group. RESULTS The pregnancy rates, multiple birth rates, abortion rates, ectopic pregnancy rates, gestational age at delivery, preterm birth rates, neonatal birth weight, and take-home baby rates were similar between the previous cesarean section group and the previous vaginal delivery group. A previous cesarean section scar did not affect embryo implantation and pregnancy outcomes in IVF. In the eSET and DET groups of previous cesarean section patients, the embryo implantation rates, pregnancy rates, abortion rates, and take-home baby rates were similar. However, the rate of multiple pregnancies reached 50% in the DET group, which led to more preterm births and lower birth weight. CONCLUSIONS Elective single-embryo transfer is a well-accepted strategy to avoid multiple pregnancies and improve the obstetric and neonatal outcomes of singleton pregnancy in IVF patients with a previous cesarean section. PMID:27636504

  6. Production of diabetic offspring using cryopreserved epididymal sperm by in vitro fertilization and intrafallopian insemination techniques in transgenic pigs.

    PubMed

    Umeyama, Kazuhiro; Honda, Kasumi; Matsunari, Hitomi; Nakano, Kazuaki; Hidaka, Tatsuro; Sekiguchi, Keito; Mochizuki, Hironori; Takeuchi, Yasuhiro; Fujiwara, Tsukasa; Watanabe, Masahito; Nagaya, Masaki; Nagashima, Hiroshi

    2013-12-17

    Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains that model human diseases. However, production of numerous cloned disease model pigs by SCNT for large-scale experiments is impractical due to its complexity and inefficiency. In the present study, we aimed to establish an efficient procedure for proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and treated with insulin to allow for normal growth to maturity, at which point epididymal sperm could be collected for cryopreservation. In vitro fertilization and intrafallopian insemination using the cryopreserved epididymal sperm resulted in diabetes model transgenic offspring. These results suggest that artificial reproductive technology using cryopreserved epididymal sperm could be a practical option for proliferation of genetically modified disease model pigs. PMID:23979397

  7. Production of Diabetic Offspring Using Cryopreserved Epididymal Sperm by In Vitro Fertilization and Intrafallopian Insemination Techniques in Transgenic Pigs

    PubMed Central

    UMEYAMA, Kazuhiro; HONDA, Kasumi; MATSUNARI, Hitomi; NAKANO, Kazuaki; HIDAKA, Tatsuro; SEKIGUCHI, Keito; MOCHIZUKI, Hironori; TAKEUCHI, Yasuhiro; FUJIWARA, Tsukasa; WATANABE, Masahito; NAGAYA, Masaki; NAGASHIMA, Hiroshi

    2013-01-01

    Abstract Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains that model human diseases. However, production of numerous cloned disease model pigs by SCNT for large-scale experiments is impractical due to its complexity and inefficiency. In the present study, we aimed to establish an efficient procedure for proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and treated with insulin to allow for normal growth to maturity, at which point epididymal sperm could be collected for cryopreservation. In vitro fertilization and intrafallopian insemination using the cryopreserved epididymal sperm resulted in diabetes model transgenic offspring. These results suggest that artificial reproductive technology using cryopreserved epididymal sperm could be a practical option for proliferation of genetically modified disease model pigs. PMID:23979397

  8. The value of the cervical score in monitoring ovulation induction for in vitro fertilization: a prospective double-blind study.

    PubMed

    Oelsner, G; Pan, S B; Barnea, E R; Boyers, S P; Tarlatzis, B C; DeCherney, A H

    1986-12-01

    The present study was designed to determine the accuracy of the cervical score for the timing of human chorionic gonadotropin (hCG) administration in in vitro fertilization patients compared to the timing of hCG administration based upon estradiol and ultrasound measurements. Forty-two patients undergoing ovulation induction with human menopausal gonadotropin (hMG)/hCG for in vitro fertilization were studied. A total of 192 cervical scores was obtained together with plasma estradiol levels and pelvic sonography. hCG was given based on estradiol and ultrasound results without knowing the cervical score. An independent decision to give hCG was made based on the cervical score only following a mean of 2.5 days of a cervical score of 9-12 without the examiner's knowledge of estradiol and ultrasound results. This decision was theoretic and was not considered for the actual timing of hCG. In 16 (38%) of the cases this decision coincided with that based on estradiol and ultrasound. In 24 (57%) of the cases there was a 1-day gap. A direct correlation was found among the cervical score, the plasma estradiol concentration, and the mean diameter of the two leading follicles (P less than 0.001). The overall distribution of the cervical score grouped according to estradiol levels (less than 200, 201-400, and greater than 401 pg/ml) was significantly different (P less than 0.001). The proportion of "mature" cervical scores (score of 11-12) in the three different estradiol groups was 15.6, 34.5, and 68.0%, respectively. In conclusion, independent decisions to give hCG based on cervical score or estradiol and ultrasound were coincident +/- 1 day in 95% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Effects of sera, hormones and granulosa cells added to culture medium for in-vitro maturation, fertilization, cleavage and development of bovine oocytes.

    PubMed

    Fukui, Y; Ono, H

    1989-07-01

    Sera (fetal calf serum: FCS; and oestrous cow serum: ECS), hormones (2.5 FSH micrograms/ml + 5 micrograms LH/ml + 1 microgram oestradiol/ml) and granulosa cells (5 x 10(6)/ml) were added to culture medium to determine the frequencies of in-vitro maturation, fertilization, cleavage (2- to 8-cell) and development into blastocysts of bovine follicular oocytes. The maturation rates after 24 h in culture were not significantly different among the three factors tested (56-72%). The fertilization rates were significantly affected by serum type and the addition of granulosa cells. FCS gave significantly higher rates of fertilization (57-71%) than did ECS (34-52%), but the proportions of polyspermic fertilization were significantly higher in the former (8-19%) than in the latter (2-3%). The addition of hormones did not affect fertilization, cleavage and development. Neither type of serum affected cleavage and development. The highest rates of blastocyst formation were obtained when granulosa cells alone were added (FCS, 17%; ECS, 16%). The cell numbers of the blastocysts obtained were 100-150, similar to those of blastocysts developed in vivo. Transfer of 6 blastocysts to 3 cows resulted in 1 pregnancy. The present results indicate that the co-culture with granulosa cells is the most important factor for in-vitro fertilization to development into blastocysts of bovine oocytes matured in vitro. PMID:2760879

  10. Effects of sera, hormones and granulosa cells added to culture medium for in-vitro maturation, fertilization, cleavage and development of bovine oocytes.

    PubMed

    Fukui, Y; Ono, H

    1989-07-01

    Sera (fetal calf serum: FCS; and oestrous cow serum: ECS), hormones (2.5 FSH micrograms/ml + 5 micrograms LH/ml + 1 microgram oestradiol/ml) and granulosa cells (5 x 10(6)/ml) were added to culture medium to determine the frequencies of in-vitro maturation, fertilization, cleavage (2- to 8-cell) and development into blastocysts of bovine follicular oocytes. The maturation rates after 24 h in culture were not significantly different among the three factors tested (56-72%). The fertilization rates were significantly affected by serum type and the addition of granulosa cells. FCS gave significantly higher rates of fertilization (57-71%) than did ECS (34-52%), but the proportions of polyspermic fertilization were significantly higher in the former (8-19%) than in the latter (2-3%). The addition of hormones did not affect fertilization, cleavage and development. Neither type of serum affected cleavage and development. The highest rates of blastocyst formation were obtained when granulosa cells alone were added (FCS, 17%; ECS, 16%). The cell numbers of the blastocysts obtained were 100-150, similar to those of blastocysts developed in vivo. Transfer of 6 blastocysts to 3 cows resulted in 1 pregnancy. The present results indicate that the co-culture with granulosa cells is the most important factor for in-vitro fertilization to development into blastocysts of bovine oocytes matured in vitro.

  11. Parabolic trend in endometrial thickness at embryo transfer in in vitro fertilization/intracytoplasmic sperm injection cases with clinical pregnancy evidence.

    PubMed

    Lamanna, Giuseppina; Scioscia, Marco; Lorusso, Filomenamila; Serrati, Giuseppe; Selvaggi, Luigi E; Depalo, Raffaella

    2008-10-01

    Sonographic measurement of endometrial thickness at embryo transfer is thought to be a good predictor of the success of in vitro fertilization/intracytoplasmic sperm injection cycles because the clinical pregnancy rate increases as the endometrium thickens. Nevertheless, a retrospective analysis of a study population of 606 patients showed a decrease of clinical pregnancy rates in the setting of extreme endometrial thicknesses.

  12. Comparison of the in vitro fertilization rate by human sperm capacitated by multiple-tube swim-up and Percoll gradient centrifugation.

    PubMed

    Tanphaichitr, N; Agulnick, A; Seibel, M; Taymor, M

    1988-06-01

    Two sperm preparation methods, a multiple-tube swim-up and Percoll-gradient centrifugation, were employed in our human in vitro fertilization program. The fertilization rate of these two sperm preparation methods was compared when they were employed in semen samples of less than 60 million motile sperm/ml. The results described here suggest that both of these methods gave a similar fertilization rate in these semen samples, i.e., 72 +/- 8% for the Percoll-gradient centrifugation method and 66 +/- 8% for the multiple-tube swim-up method.

  13. /sup 125/I-labeled radioimmunoassay kits for progesterone evaluated for use in an in vitro fertilization program

    SciTech Connect

    Blight, L.F.; White, G.H.

    1983-06-01

    We have evaluated two commercially available /sup 125/I radioimmunoassay kits (Diagnostic Products Corp., DPC; and Radioassay Systems Laboratories, RSL) for measurement of serum or plasma progesterone, to determine their suitability for use in in vitro fertilization programs. Both kits were suitably rapid for program requirements. Results by both were linear with concentration up to 60 nmol/L, and both had acceptable lower detection limits of 0.3 nmol/L. Kit-determined progesterone concentrations (y) for 100 patients' samples correlated well with results by our existing 3H radioimmunoassay method (y . 1.11x + 0.2, r . 0.965 for the DPC kit; y . 1.01x + 1.4, r . 0.974 for the RSL kit). Mean analytical recovery for the RSL kit was 116%, that for the DPC kit, 202%. Within-batch precision, expressed as the mean CV for three concentrations of progesterone, was 6.5% for the RSL kit, and 16.4% for the DPC kit; between-day CV was 8.1% for the RSL kit, 17.7% for the DPC kit. We conclude that the RSL kit provides a rapid, precise, and accurate assay for serum progesterone, suitable for use in a fertilization program, but do not recommend the DPC kit for either this purpose or the more general purpose of tracking menstrual cycles.

  14. The effect of FF-MAS on porcine cumulus-oocyte complex maturation, fertilization and pronucleus formation in vitro.

    PubMed

    Faerge, Inger; Strejcek, Frantisek; Laurincik, Jozef; Rath, Detlef; Niemann, Heiner; Schellander, Karl; Rosenkranz, Christine; Hyttel, Poul Maddox; Grøndahl, Christian

    2006-08-01

    Follicular fluid meiosis-activating sterol (FF-MAS) has been isolated from the follicular fluid (FF) of several species including man. FF-MAS increases the quality of in vitro oocyte maturation, and thus the developmental potential of oocytes exposed to FF-MAS during in vitro maturation is improved. The aim of the present study was to investigate the effects of FF-MAS on porcine oocyte maturation and pronucleus formation in vitro. Porcine cumulus-oocyte complexes (COCs) were isolated from abattoir ovaries and in vitro matured for 48 h in NCSU 37 medium supplemented with 1 mg/l cysteine, 10 ng/ml epidermal growth factor and 50 microM 2-mercaptoethanol with or without 10% porcine follicular fluid (pFF). For the first 22 h, 1 mM db-cAMP and 10 I.E PMSG/hCG was added. The medium was supplemented with 1 microM, 3 microM, 10 microM, 30 microM or 100 microM FF-MAS dissolved in ethanol. After maturation the COCs were denuded mechanically using a fine glass pipette under constant pH and in vitro fertilized with fresh semen (5 x 10(5) spermatozoa/ml). The presumptive zygotes were evaluated 18 h after fertilization. The addition of pFF increased the monospermic as well as the polyspermic penetration of oocytes. In the absence of pFF, the addition of FF-MAS decreased the polyspermic penetration rate, whereas FF-MAS in combination with pFF decreased monospermic and increased polyspermic penetration. The degeneration rate of ova decreased in the presence of FF-MAS irrespective of the presence or absence of pFF. In the absence of pFF, FF-MAS at 3-10 microM increased the number of zygotes with advanced maternal pronuclear stages. In supraphysiological doses, i.e. 30-100 microM, FF-MAS dose-dependently and reversibly inhibited nuclear maturation in the absence of pFF.

  15. Effects of norethindrone on gonadotropin and ovarian steroid secretion when used for cycle programming during in vitro fertilization.

    PubMed

    Anderson, R E; Stein, A L; Paulson, R J; Stanczyk, F Z; Vijod, A G; Lobo, R A

    1990-07-01

    Norethindrone (NET) has been used for cycle programming and may result in attenuated responses to controlled ovarian hyperstimulation. The effects of NET on gonadotropin secretion, its bioavailability to the ovary, and its effect on ovarian steroidogenesis in vivo and in vitro were assessed. Endogenous secretion of luteinizing hormone and follicle-stimulating hormone was attenuated by 59% and 50%, respectively, after 2 weeks of orally administered NET. Twelve hours after a single 10-mg oral dose, significant levels of NET were measured in samples of peripheral (8.8 +/- 1.9 ng/mL) and ovarian venous blood (10.5 +/- 3.1 ng/mL), follicular fluid (7.1 +/- 2.1 ng/mL), and homogenates of ovarian tissue (8.0 +/- 0.6 ng/g). Furthermore, NET was detectable in follicular fluid 2 weeks after its withdrawal (863 +/- 149 pg/mL). However, there were no effects of NET on follicular fluid levels of estradiol and progesterone in vivo or on luteinized granulosa cell steroidogenesis in vitro. We conclude that when used for cycle programming in in vitro fertilization, NET does not inhibit ovarian steroidogenesis but does affect the hypothalamic-pituitary axis.

  16. From Embryos to Adults: A DOHaD Perspective on In Vitro Fertilization and Other Assisted Reproductive Technologies.

    PubMed

    Feuer, Sky; Rinaudo, Paolo

    2016-01-01

    Human in vitro fertilization (IVF) as a treatment for infertility is regarded as one of the most outstanding accomplishments of the 20th century, and its use has grown dramatically since the late 1970s. Although IVF is considered safe and the majority of children appear healthy, reproductive technologies have been viewed with some skepticism since the in vitro environment deviates substantially from that in vivo. This is increasingly significant because the Developmental Origins of Health and Disease (DOHaD) hypothesis has illuminated the sensitivity of an organism to its environment at critical stages during development, including how suboptimal exposures restricted specifically to gamete maturation or the preimplantation period can affect postnatal growth, glucose metabolism, fat deposition, and vascular function. Today, some of the physiological metabolic phenotypes present in animal models of IVF have begun to emerge in human IVF children, but it remains unclear whether or not in vitro embryo manipulation will have lasting health consequences in the offspring. Our expanding knowledge of the DOHaD field is fueling a paradigm shift in how disease susceptibility is viewed across the life course, with particular emphasis on the importance of collecting detailed exposure information, identifying biomarkers of health, and performing longitudinal studies for any medical treatment occurring during a developmentally vulnerable period. As IVF use continues to rise, it will be highly valuable to incorporate DOHaD concepts into the clinical arena and future approaches to public health policy. PMID:27517965

  17. From Embryos to Adults: A DOHaD Perspective on In Vitro Fertilization and Other Assisted Reproductive Technologies

    PubMed Central

    Feuer, Sky; Rinaudo, Paolo

    2016-01-01

    Human in vitro fertilization (IVF) as a treatment for infertility is regarded as one of the most outstanding accomplishments of the 20th century, and its use has grown dramatically since the late 1970s. Although IVF is considered safe and the majority of children appear healthy, reproductive technologies have been viewed with some skepticism since the in vitro environment deviates substantially from that in vivo. This is increasingly significant because the Developmental Origins of Health and Disease (DOHaD) hypothesis has illuminated the sensitivity of an organism to its environment at critical stages during development, including how suboptimal exposures restricted specifically to gamete maturation or the preimplantation period can affect postnatal growth, glucose metabolism, fat deposition, and vascular function. Today, some of the physiological metabolic phenotypes present in animal models of IVF have begun to emerge in human IVF children, but it remains unclear whether or not in vitro embryo manipulation will have lasting health consequences in the offspring. Our expanding knowledge of the DOHaD field is fueling a paradigm shift in how disease susceptibility is viewed across the life course, with particular emphasis on the importance of collecting detailed exposure information, identifying biomarkers of health, and performing longitudinal studies for any medical treatment occurring during a developmentally vulnerable period. As IVF use continues to rise, it will be highly valuable to incorporate DOHaD concepts into the clinical arena and future approaches to public health policy. PMID:27517965

  18. Programmed oocyte retrieval: clinical and biological effects of oral contraceptives administered before in vitro fertilization.

    PubMed

    Mashiach, S; Dor, J; Goldenberg, M; Shalev, J; Levran, D; Rudak, E; Nebel, L; Goldman, B; Blankstein, J; Ben-Rafael, Z

    1989-06-01

    We have prospectively compared two regimens of suppression of the hypothalamic-pituitary-ovarian axis by oral contraceptives (OCs) for 15 or 30 days and two ovarian stimulation protocols. The latent phase, which represents a period of ovarian insensitivity, was prolonged and directly correlated to the duration of suppression. Thirty days' suppression, compared with 15 days', resulted in the cancellation of more cycles and a lower fertilization and pregnancy percentage. No significant increase in either serum progesterone or luteinizing hormone was noted in suppressed cycles. It is concluded that if programming is desired, OCs should be used for the shortest period possible. The variation in the length of the follicular phase indicates that there is a different 'fixed' day for retrieval for each suppression-stimulation protocol and this day should be established prospectively.

  19. Influence of co-culture with denuded oocytes during in vitro maturation on fertilization and developmental competence of cumulus-enclosed porcine oocytes in a defined system.

    PubMed

    Appeltant, Ruth; Somfai, Tamás; Kikuchi, Kazuhiro; Maes, Dominiek; Van Soom, Ann

    2016-04-01

    Co-culture of cumulus-oocyte complexes (COCs) with denuded oocytes (DOs) during in vitro maturation (IVM) was reported to improve the developmental competence of oocytes via oocyte-secreted factors in cattle. The aim of the present study was to investigate if addition of DOs during IVM can improve in vitro fertilization (IVF) and in vitro culture (IVC) results for oocytes in a defined in vitro production system in pigs. The maturation medium was porcine oocyte medium supplemented with gonadotropins, dbcAMP and β-mercaptoethanol. Cumulus-oocyte complexes were matured without DOs or with DOs in different ratios (9 COC, 9 COC+16 DO and 9 COC+36 DO). Consequently; oocytes were subjected to IVF as intact COCs or after denudation to examine if DO addition during IVM would affect cumulus or oocyte properties. After fertilization, penetration and normal fertilization rates of zygotes were not different between all tested groups irrespective of denudation before IVF. When zygotes were cultured for 6 days, no difference could be observed between all treatment groups in cleavage rate, blastocyst rate and cell number per blastocyst. In conclusion, irrespective of the ratio, co-culture with DOs during IVM did not improve fertilization parameters and embryo development of cumulus-enclosed porcine oocytes in a defined system.

  20. [Results of diagnostic hysteroscopies in an in vitro fertilization/embryo transfer program].

    PubMed

    Bordt, J; Belkien, L; Vancaillie, T; Stening, C; Schneider, H P

    1984-12-01

    Twentysix sterile women were examined during the development and growth phase of the follicle, using a microhysteroscope manufactured by Karl Storz, under local anaesthesia. All these patients had been included in the in-vitro fertilisation/embryo transfer (IVF/ET) programme because their oviducts were either missing or had been irreparably damaged. Pathological findings were present in 7 patients. Cervical conditions were normal in 22 patients, and intrauterine conditions were without any abnormal findings in 23 patients. The clinical relevance of the findings with regard to embryo transfer and implantation are discussed. It is concluded that hysteroscopy not only occupies an important rank in the general diagnosis of sterility, but is also of considerable importance in preparatory work for embryo transfer.

  1. The use of time lapse photography in an in vitro fertilization programme for better selection for embryo transfer.

    PubMed

    Kovačič, Borut; Hojnik, Nina; Vlaisavljević, Veljko

    2014-01-01

    The time lapse photography is not a new method for assessing the dynamics of early embryo development in vitro. It has been used many times in the past for studying cleavages and blastulation of embryos of various animal species. However, this technique became available for routine use in an human in vitro fertilization (IVF) programme only a couple years ago and it becomes more and more popular today. The new time lapse systems are using modified microscopes which are positioned within the incubators. The observation of embryos does not need the opening of incubators. By sequential photographing of each embryo separately with camera of low intensity illumination, more than 1400 pictures of embryo are made. All these pictures are collected together and transformed into a short movie with software. This system offers the observation of dynamics of embryo development. The studies, which have used a time lapse technique for studying embryo development, revealed that the timing between different events can be used for predicting its developmental potential. In this paper the advantages and drawbacks of time lapse photography is precisely described. An overview through the published papers analyzing the dynamics of human embryo development from the zygote toward blastocyst is done and new timing parameters for grading zygotes, early embryos and blastocysts are analyzed.

  2. Transrectal electroejaculation combined with in-vitro fertilization: effective treatment of anejaculatory infertility due to spinal cord injury.

    PubMed

    Brinsden, P R; Avery, S M; Marcus, S; Macnamee, M C

    1997-12-01

    Infertility due to spinal cord injury (SCI) in young men is a frequent complication of their injury. When the simpler methods of management of the erectile and ejaculatory dysfunction that invariably follow the more severe types of SCI are not effective, then semen production by transrectal electroejaculation (TREE) combined with in-vitro fertilization (IVF) and embryo transfer is effective. A retrospective analysis is presented of data on the treatment and outcome of 35 couples who wished to have a family but in whom the male partner had suffered SCI. These 35 couples had 71 attempts at IVF with spermatozoa obtained following TREE. Normal fertilization and cleavage of the embryos occurred in 48.2% of the oocytes. Fresh embryos were transferred in 54 cycles and frozen-thawed embryos in 14 cycles. In all, 18 clinical pregnancies were achieved in 54 fresh and 14 frozen embryo transfer cycles, with a live birth rate of 16.5% (14/85) per treatment cycle started, 20.6% (14/68) per transfer cycle and 40.0% (14/35) per couple who started treatment, in a mean of 1.9 transfer cycles. We conclude that TREE combined with IVF and embryo transfer is an effective treatment for the infertility problems associated with SCI.

  3. [The analysis of physicians' work: announcing the end of attempts at in vitro fertilization].

    PubMed

    Santiago-Delefosse, M; Cahen, F; Coeffin-Driol, C

    2003-01-01

    The purpose of this empirical study is to analyze modalities of announcing the end of attempts at in vitro ferti-lization to women who, for various reasons, were not able to have a child after several trials. What are the problems physicians face when, in the course of their work, they make these announcements? How do they give (or not give) support to these women who have placed so much hope in this technique? These are some of the questions that led the authors to conduct this empirical study within the framework of a clinical and qualitative approach to work psychology. Within this framework, work is conceptualised as a complex activity that involves the subject, both bodily and through his various modes of socialisation. The field of clinical and quali-tative approach to work psychology situations focuses on different ways of expressing distress related to contradictory work demands, as the activity is being performed; it also focuses on those creative processes used by the subject to cope with those internal and external conflicts that hinder task performance. A review of the literature and preliminary observations led us to postulate that the problems physicians are faced with when they announce the end of attempts at in vitro fertilisation (IVF) are linked to several conflicts between work values (that are specific to the medical world) and the recognition of work failure: termination of attempts at IVF. The popu-lation that participated in this research project belongs to a network of private practitioners who work with the in-house team of a Parisian clinic. But the group is not uniform and some physicians perform IVFs more frequently than others. Our qualitative study involved 10 semi-directive interviews of approximately 1 1/2 hours each, which were recorded and transcribed. Initial instructions focused on a concrete description of situations of abandonment of attempts at IVF, in terms of their preparation, development, and the way they are experienced

  4. Successful treatment of infertility due to anejaculation with in vitro fertilization and embryo transfer: a report of two cases.

    PubMed

    Nagai, S; Kasai, T; Ogawa, K; Mizuno, K; Ohta, S; Hoshi, K

    1998-03-01

    We investigated the efficacy of assisted ejaculation in combination with in vitro fertilization and embryo transfer (IVF-ET) in two couples with infertility due to anejaculation. Case 1 was in an anejaculatory condition associated with a spinal cord-injury. We performed intrauterine insemination (IUI) three times using motile sperm obtained by intrathecal injection of neostigmine, but no pregnancy followed. The couple then entered IVF-ET. The semen volume obtained by assisted ejaculation was 4.5 ml. The sperm count was 33 x 10(6)/ml with 33% motility. At 35 weeks of gestation she delivered three healthy babies by cesarean section. Case 2 was in an unexplained retrograde ejaculatory condition. We performed IUI eight times before changing to IVF-ET. We then used sperm obtained from urine or sperm recovered from the bladder after administration of human tubal fluid, but no pregnancy followed. At the fifth IVF-ET attempt, using sperm obtained from urine, we succeeded. We prepared the sperm by the Percoll gradient method. The sperm count was 36 x 10(6)/ml with 64% motility. At 39 weeks of gestation she delivered a healthy baby. Deficient seminal quality contributes to the decreased fertility potential in patients with anejaculation. With the use of IVF-ET methods, pregnancies can be achieved in these couples.

  5. Lack of transmission of murine norovirus to mice via in vitro fertilization, intracytoplasmic sperm injection, and ovary transplantation.

    PubMed

    Raspa, Marcello; Mahabir, Esther; Fray, Martin; Volland, Ruth; Scavizzi, Ferdinando

    2016-07-15

    Since its discovery in 2003, murine norovirus (MNV) is still endemic in many rodent animal facilities. Our aim was to determine the risk of transmission of MNV (91% homology to MNV3) to embryo recipients and pups via assisted reproductive technologies, especially those which compromise the integrity of the zona pellucida. In vitro fertilization (IVF), assisted in vitro fertilization (AIVF) with reduced glutathione, intracytoplasmic sperm injection, and ovary transplantation were performed. Murine norovirus was detected by qualitative and quantitative reverse transcription polymerase chain reaction. After natural infection of immunocompetent C57BL/6NTacCnrm and immunodeficient athymic nude mice with MNV, the mesenteric lymph nodes, small intestine, spleen, liver, lung, brain, ovary, and testis were infected at specific intervals for more than a 1-year period. At Week 12, the number of viral genomes per milligram of gonad from both strains was 20 to 50. Murine norovirus strictly adhered to spermatozoa collected from infected mice because three washes did not remove MNV from the sperm. After using MNV-positive sperm for IVF, AIVF, and intracytoplasmic sperm injection, 27 to 30 genomes were detected in IVF (n = 100) and AIVF (n = 100) embryos from both mouse strains. Approximately 87% of MNV detected in these embryos was found in the zona pellucida. However, all embryo transfer recipients, pups, and ovary recipients were MNV-negative. The results indicate that manipulation of the germplasm through assisted reproductive technologies did not lead to transmission of MNV to mice. This may be because of the absence of an infectious dose or failure of the MNV strain to replicate effectively in developing embryos and the reproductive tract. PMID:26972226

  6. Lack of transmission of murine norovirus to mice via in vitro fertilization, intracytoplasmic sperm injection, and ovary transplantation.

    PubMed

    Raspa, Marcello; Mahabir, Esther; Fray, Martin; Volland, Ruth; Scavizzi, Ferdinando

    2016-07-15

    Since its discovery in 2003, murine norovirus (MNV) is still endemic in many rodent animal facilities. Our aim was to determine the risk of transmission of MNV (91% homology to MNV3) to embryo recipients and pups via assisted reproductive technologies, especially those which compromise the integrity of the zona pellucida. In vitro fertilization (IVF), assisted in vitro fertilization (AIVF) with reduced glutathione, intracytoplasmic sperm injection, and ovary transplantation were performed. Murine norovirus was detected by qualitative and quantitative reverse transcription polymerase chain reaction. After natural infection of immunocompetent C57BL/6NTacCnrm and immunodeficient athymic nude mice with MNV, the mesenteric lymph nodes, small intestine, spleen, liver, lung, brain, ovary, and testis were infected at specific intervals for more than a 1-year period. At Week 12, the number of viral genomes per milligram of gonad from both strains was 20 to 50. Murine norovirus strictly adhered to spermatozoa collected from infected mice because three washes did not remove MNV from the sperm. After using MNV-positive sperm for IVF, AIVF, and intracytoplasmic sperm injection, 27 to 30 genomes were detected in IVF (n = 100) and AIVF (n = 100) embryos from both mouse strains. Approximately 87% of MNV detected in these embryos was found in the zona pellucida. However, all embryo transfer recipients, pups, and ovary recipients were MNV-negative. The results indicate that manipulation of the germplasm through assisted reproductive technologies did not lead to transmission of MNV to mice. This may be because of the absence of an infectious dose or failure of the MNV strain to replicate effectively in developing embryos and the reproductive tract.

  7. Follicular fluid and urinary concentrations of phthalate metabolites among infertile women and associations with in vitro fertilization parameters.

    PubMed

    Du, Yao-Yao; Fang, Yue-Li; Wang, Yi-Xin; Zeng, Qiang; Guo, Na; Zhao, Hua; Li, Yu-Feng

    2016-06-01

    Evidence from toxicological studies has demonstrated that phthalates can lead to reduced fertility through effects on folliculogenesis, oocyte maturation and embryonic development, but human data are limited. Concentrations of eight phthalate metabolites in 110 follicular fluid (FF) and urine samples collected from 112 women attending an infertility clinic in Wuhan, China were quantified, and correlations between paired matrices were explored. Associations between metabolite concentrations and in vitro fertilization (IVF) parameters were evaluated with multivariable models. Six metabolites were detected in >72.73% of the FF samples. MEHP and MBP were the dominant metabolites with a median level of 2.80 and 2.05ng/mL, respectively. Significant correlations between the two matrices, urine and FF, were found for MEP (rs=0.44), and MBP (rs=0.22). FF and urinary metabolite concentrations were not associated with any IVF parameters. However, given the prevalence of phthalates exposure, further work is needed to elucidate the potential hazard on female reproduction. PMID:27067915

  8. Attempts at in vitro fertilization and culture of in vitro matured oocytes in sei ( Balaenoptera borealis) and Bryde's ( B. edeni) whales.

    PubMed

    Bhuiyan, M M U; Suzuki, Y; Watanabe, H; Matsuoka, K; Fujise, Y; Ishikawa, H; Ohsumi, S; Fukui, Y

    2009-02-01

    The cumulus-oocyte-complexes (COCs) recovery rates with respect to reproductive status per sei (Balaenoptera borealis) and Bryde's (B. edeni) whales were determined in Experiment 1. The number of COCs recovered ranged from 16.0 to 30.6 and from 6.7 to 26.8 per sei and Bryde's whales, respectively. The effects of COCs grades and protein supplementation in embryo culture medium on development of in vitro fertilized (IVF) embryos were evaluated in sei and Bryde's whales in Experiment 2. The COCs were classified into either Grade A (COCs with five or more layers of compact cumulus cells) or Grade B (COCs with less than five layers of compact or expanded cumulus cells) before being cultured for IVM. The cleavage (12.0 to 19.5%), 4-cell (8.0 to 12.0%) and 8-cell (4.0 to 8.0%) formation rates in sei whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either fetal whale serum (FWS)- or bovine serum albumin (BSA)-supplemented medium. The cleavage (4.0 to 14.8%), 4-cell (0.0 to 7.5%) and 8-cell (0.0 to 2.6%) formation rates in Bryde's whales did not vary significantly between embryos derived from either grade A or B oocytes and between embryos cultured in either FWS- or BSA-supplemented medium. The grade B oocytes cultured in FWS-supplemented medium developed to morula stage (1.1%) in sei whales. In conclusion, the present study indicates that IVF in sei whales is possible to achieve cleaved embryos developing to morula stage. This is the first in vitro embryo production attempt in sei and Bryde's whales.

  9. Porcine embryo production following in vitro fertilization and intracytoplasmic sperm injection from vitrified immature oocytes matured with a granulosa cell co-culture system.

    PubMed

    Casillas, Fahiel; Ducolomb, Yvonne; Lemus, Ana E; Cuello, Cristina; Betancourt, Miguel

    2015-10-01

    This study was designed to evaluate the capacity of vitrified-warmed porcine immature oocytes to mature and to be fertilized using in vitro fertilization or intracytoplasmic sperm injection, and to determine the subsequent embryo development. Immature oocytes were vitrified using ethylene glycol and dimethylsulphoxide as cryoprotectants and the Cryolock method. After warming oocytes were cultured 44 h for maturation. Oocytes were randomly distributed in three treatment groups and subjected to in vitro fertilization (Experiment 1) or intracytoplasmic sperm injection (Experiment 2) procedures. The results indicate that the embryo development was higher in denuded oocytes co-cultured with granulosa cells (NkO-CC group) fertilized by in vitro fertilization or intracytoplasmic sperm injection compared to cumulus-cell oocyte complexes (COCs group), showing no significant differences with control. Vitrified denuded oocytes matured with a co-culture system NkO-CC group, displayed higher cleavage rate and blastocyst production than vitrified COCs group. Blastocysts were successfully obtained after IVF and ICSI procedures; however, the development to the blastocyst stage was better after IVF. These results show that the vitrification-warming media, the employment of a granulosa cell co-culture system and the Cryolock method during vitrification, increased the nuclear and cytoplasmic maturation of vitrified porcine immature oocytes. Further experiments are required to enhance porcine embryo production after vitrification.

  10. [Surgical pelviscopy in treatment of extrauterine pregnancy following in vitro fertilization and spontaneous conception].

    PubMed

    Fiedler, K; Krüsmann, G; Würfel, W

    1991-08-01

    We report on our experience with surgical pelviscopy in the treatment of ectopic pregnancies after in-vitro fertilisation and, in comparison, during spontaneous cycles and during cycles with ovarian hyperstimulations and IUI. From 1.1. 1988 to 31.12.1990, 54 patients underwent this operation, 25 of these having undergone the IVF programme. All IVF patients had an extensive history of tubal sterility. Therefore, we found difficult anatomic conditions during the operation in most cases. Furthermore, the special conditions of the IVF treatment itself (e.g., multiple transfer of embryos into the uterus) caused a higher incidence of complicated situations (one triple ectopic, three ovarian ectopic pregnancies and one simultaneous pregnancy). Firstly, our experience leads to the conclusion, that surgical pelviscopy can be used for management of more complicate problems and can be regarded, so far, as an alternative to laparotomy. Secondly, laparotomy should not be replaced by pelviscopy in difficult situations (e.g., ovarian ectopics pregnancies). Surgical pelviscopy reduces the trauma of treatment, a consideration, which is very important especially for IVF patients. Because of the extraordinary problems related to ectopic pregnancies after IVF, we are at present sceptical about the use of other methods (e.g., instillation of prostaglandins or methotrexate) for these patients. Subsequently, 19 patients in the IVF group conceived with an intrauterine pregnancy after repeated IVF treatment. In the second group, 8 spontaneous intrauterine conceptions have occurred since. PMID:1834513

  11. The "Test-Tube" Generation: Parent-Child Relationships and the Psychological Well-Being of In Vitro Fertilization Children at Adolescence.

    ERIC Educational Resources Information Center

    Golombok, Susan; MacCallum, Fiona; Goodman, Emma

    2001-01-01

    Compared parent-child relationships and early adolescent well-being in families with children conceived by in vitro fertilization (IVF), adoptive families, and families with a naturally conceived child. Found that IVF children were functioning well and did not differ from other children in social or emotional adjustment. (Author/KB)

  12. A Comprehensive Analysis of Body Mass Index Effect on in Vitro Fertilization Outcomes

    PubMed Central

    Sarais, Veronica; Pagliardini, Luca; Rebonato, Giorgia; Papaleo, Enrico; Candiani, Massimo; Viganò, Paola

    2016-01-01

    The effect of a raised body mass index (BMI) on the outcome of assisted reproduction technology (ART) still represents a controversial issue. Even less clear is whether BMI acts with a potential detrimental effect on IVF outcomes via a deleterious effect on innate quality of oocytes or on the environmental milieu within the uterus. With the aim to better understand the mechanisms underlying the potential deleterious effect of an increased BMI on IVF outcomes, we have evaluated the effects of female BMI on number and quality of retrieved oocytes, fertilization rate, embryo score and incidences of ongoing pregnancy and live births among couples undergoing IVF in an Italian population. Data from 1602 women who underwent their first IVF cycle were retrospectively analyzed. A significantly reduced percentage of mature oocytes when comparing obese (BMI ≥ 30 kg/m2) and normal-weight patients (BMI = 18.50–24.99 kg/m2) was found. After adjusting for maternal age and other confounders, odds for ongoing pregnancy rate showed no differences across different BMI categories. However, a significant increased odds ratio (OR) could be observed for miscarriage rate in patients with BMI ≥ 25 (OR = 2.5; p = 0.04). These results should be taken into account in order to define optimal strategies for overweight and obese patients referring to ART procedures. PMID:26907340

  13. A Comprehensive Analysis of Body Mass Index Effect on in Vitro Fertilization Outcomes.

    PubMed

    Sarais, Veronica; Pagliardini, Luca; Rebonato, Giorgia; Papaleo, Enrico; Candiani, Massimo; Viganò, Paola

    2016-02-23

    The effect of a raised body mass index (BMI) on the outcome of assisted reproduction technology (ART) still represents a controversial issue. Even less clear is whether BMI acts with a potential detrimental effect on IVF outcomes via a deleterious effect on innate quality of oocytes or on the environmental milieu within the uterus. With the aim to better understand the mechanisms underlying the potential deleterious effect of an increased BMI on IVF outcomes, we have evaluated the effects of female BMI on number and quality of retrieved oocytes, fertilization rate, embryo score and incidences of ongoing pregnancy and live births among couples undergoing IVF in an Italian population. Data from 1602 women who underwent their first IVF cycle were retrospectively analyzed. A significantly reduced percentage of mature oocytes when comparing obese (BMI ≥ 30 kg/m²) and normal-weight patients (BMI = 18.50-24.99 kg/m²) was found. After adjusting for maternal age and other confounders, odds for ongoing pregnancy rate showed no differences across different BMI categories. However, a significant increased odds ratio (OR) could be observed for miscarriage rate in patients with BMI ≥ 25 (OR = 2.5; p = 0.04). These results should be taken into account in order to define optimal strategies for overweight and obese patients referring to ART procedures.

  14. Effect of caffeine on motility and vitality of sperm and in vitro fertilization of outbreed mouse in T6 and M16 media

    PubMed Central

    Nabavi, Narges; Todehdehghan, Fatemeh; Shiravi, Abdollhossein

    2013-01-01

    Background: Caffeine increases the CAMP production that stimulates spermatozoa movement. Caffeine is also used for induction of in vitro acrosome reaction in mammalian spermatozoa, an important step in achieving fertilization. Objective: The aim of this study was to assess the effect of caffeine on sperm's motility, vitality and laboratory fertilization rates in mouse in two T6 and M16 media. Materials and Methods: Epididymal mouse sperms were collected and treated by caffine in T6 and M16 media and their motility and vitality rates were evaluated. The pretreated sperms were added to oocytes in T6 and M16 media with and without caffeine and fertilization rates were recorded after 24 hours incubation. Results: Sperm's motility (81.7±1.67%) and vitality (88.7±1.33%) rates and percentage of fertilized oocytes (67.52±8.16%) in T6 medium plus caffeine compare to control group have increased and shown significant differences at p≤0.01. While the percentages of these parameters in M16 medium supplemented with caffeine were 68.3±6.01%, 78±6.11%, and 42.6±12.96 respectively and in comparison to control group (M16 without caffeine) have not shown significant differences. Conclusion: Addition of caffeine to T6 medium promotes the sperm's motility and vitality and enhances fertilization and early in vitro development of mouse embryos. This article extracted from M.Sc. thesis. (Narges Navabi) PMID:24639814

  15. Developmental competence of frozen-thawed yak (Bos grunniens) oocytes followed by in vitro maturation and fertilization.

    PubMed

    Niu, Hui-Ran; Zi, Xiang-Dong; Xiao, Xiao; Xiong, Xian-Rong; Zhong, Jin-Cheng; Li, Jian; Wang, Li; Wang, Yong

    2014-02-01

    In the present study, we examined the ability of immature germinal vesicle (GV) and subjected to in vitro matured (MII) yak oocytes to survive after cryopreservation as well as their subsequent development following in vitro maturation and fertilization. Both GV and MII oocytes were cryopreserved by using two different vitrification solutions (VS); VS-I contained 10% ethylene glycol (EG) and 10% dimethylsulfoxide (DMSO) in TCM-199 + 20% (v/v) fetal calf serum (FCS) whereas VS-II contained 40% EG + 18% Ficoll + 0.5 M sucrose in TCM-199 + 20% FCS. The percentage of oocytes found to be morphologically normal was greater (P < 0.01) in VS-I group than in VS-II group. Rates of cleavage (30.6-42.2%) and blastocyst formation (2.9-8.9%) did not differ among groups, but were lower than in unfrozen control (55.7% and 25.4%, P < 0.01). These results show that a combination of EG and DMSO or EG, Ficoll and sucrose can be used to cryopreserve yak oocytes in French straws.

  16. Moderate Ovarian Stimulation Does Not Increase the Incidence of Human Embryo Chromosomal Abnormalities in in Vitro Fertilization Cycles

    PubMed Central

    Bosch, Ernesto; Alamá, Pilar; Rubio, Carmen; Rodrigo, Lorena; Pellicer, Antonio

    2012-01-01

    Context: A high chromosomal abnormalities rate has been observed in human embryos derived from in vitro fertilization (IVF) treatments. The real incidence in natural cycles has been poorly studied, so whether this frequency may be induced by external factors, such as use of gonadotropins for ovarian stimulation, remains unknown. Design: We conducted a prospective cohort study in a University-affiliated private infertility clinic with a comparison between unstimulated and stimulated ovarian cycles in the same women. Preimplantation genetic screening by fluorescence in situ hybridization was performed in all viable d 3 embryos. Objective: The primary objective was to compare the incidence of embryo chromosomal abnormalities in an unstimulated cycle and in an ulterior moderate ovarian stimulated cycle. Secondary outcome measures were embryo quality, blastocyst rate of biopsied embryos, number of normal blastocysts per donor, type of chromosomal abnormalities, and clinical outcome. Results: One hundred eighty-five oocyte donors were initially recruited for the unstimulated cycle, and preimplantation genetic screening could be performed in 51 of them, showing 35.3% of embryo chromosomal abnormalities. Forty-six of them later completed a stimulated cycle. The sperm donor sample was the same for both cycles. The proportion of embryos displaying abnormalities in the unstimulated cycle was 34.8% (16 of 46), whereas it was 40.6% (123 of 303) in the stimulated cycle with risk difference = 5.8 [95% confidence interval (CI) = −20.6–9.0], and relative risk = 1.17 (95% CI = 0.77–1.77) (P = 0.45). When an intrasubject comparison was made, the abnormalities rate was 34.8% (95% CI = 20.5–49.1) in the unstimulated cycle and 38.2% (95% CI = 30.5–45.8) in the stimulated cycle [risk difference = 3.4 (95% CI = −17.9–11.2); P = 0.64]. No differences were observed for embryo quality and type of chromosomal abnormalities. Conclusions: Moderate ovarian stimulation in young

  17. Impact of a second insemination on the results of an in vitro fertilization-embryo transfer (IVF-ET) program.

    PubMed

    Fahmy, N W; Benoit, J; Bissonnette, F; Duchesne, C; Girard, Y; Sullivan, R

    1991-04-01

    In an attempt to increase the fertilization and pregnancy rates in our program, a second insemination was carried out when the first insemination yielded fewer than two fertilized oocytes. One hundred eighty consecutive patients were studied retrospectively and thirty-four required second insemination, 35% of them by donor semen. Fifty-five and nine-tenths percent of the patients had at least one fertilized oocyte for embryo transfer, but only 21.9% of the oocytes exposed to a second insemination were fertilized. No pregnancy resulted from the transfer of oocytes fertilized by the second insemination. There were no significant correlations between the success of fertilization after a second insemination and the number of oocytes retrieved, the protocol for the induction of superovulation, or the age of the female patient. Considering that the first insemination was done at a variable time after oocyte retrieval to allow oocyte maturation, we expected all oocytes to be mature at the time of first insemination and we considered the possibility of delayed fertilization as negligible since second insemination was done at least 24-30 hr after oocyte retrieval. Even though a second insemination provides further hope for the patient, by yielding additional fertilized oocytes for embryo transfer, its main value is that it may provide additional information about male fertility.

  18. The Role of Interleukin-18 in Serum and Follicular Fluid during In Vitro Fertilization and Intracytoplasmic Sperm Injection

    PubMed Central

    Fuhs, Corinna; Salmassi, Ali; Hedderich, Jürgen; Maass, Nicolai; Elessawy, Mohamed; Schmutzler, Andreas Gerd; Eckmann-Scholz, Christel

    2016-01-01

    Cytokines are key modulators of the immune system and play an important role in the ovarian cycle. IL-18 levels in serum and follicular fluid were analyzed in women undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment. The cohort study group consisted of 90 women, who were undergoing IVF or ICSI. The body mass index (BMI) was determined in all patients; IL-18 levels were measured in follicular fluid and serum. IL-18 levels in serum were significantly higher than those in follicular fluid. The median level in serum was 162.75 (80.21) pg/mL and that in follicular fluid, 138.24 (91.78) pg/mL. Women undergoing IVF treatment had lower IL-18 levels in serum (median, 151.19 (90.73) pg/mL) than those treated with ICSI (median, 163.57 (89.97) pg/mL). The correlation between IL-18 levels in serum and BMI was statistically significant, as well as the correlation between IL-18 levels in follicular fluid and ovarian stimulation response (p = 0.003). IL-18 was correlated with the response to ovarian stimulation and was the reason for successful pregnancy after IVF or ICSI treatment. Among other cytokines, IL-18 appears to be a promising prognostic marker of success in reproductive treatment and should be evaluated as such in further prospective studies. PMID:27747236

  19. Depletion of human serum albumin in embryo culture media for in vitro fertilization using monolithic columns with immobilized antibodies.

    PubMed

    Tarasova, Irina A; Lobas, Anna A; Černigoj, Urh; Solovyeva, Elizaveta M; Mahlberg, Barbara; Ivanov, Mark V; Panić-Janković, Tanja; Nagy, Zoltan; Pridatchenko, Marina L; Pungor, Andras; Nemec, Blaž; Vidic, Urška; Gašperšič, Jernej; Krajnc, Nika Lendero; Vidič, Jana; Gorshkov, Mikhail V; Mitulović, Goran

    2016-09-01

    Affinity depletion of abundant proteins such as HSA is an important stage in routine sample preparation prior to MS/MS analysis of biological samples with high range of concentrations. Due to the charge competition effects in electrospray ion source that results in discrimination of the low-abundance species, as well as limited dynamic range of MS/MS, restricted typically by three orders of magnitude, the identification of low-abundance proteins becomes a challenge unless the sample is depleted from high-concentration compounds. This dictates a need for developing efficient separation technologies allowing fast and automated protein depletion. In this study, we performed evaluation of a novel immunoaffinity-based Convective Interaction Media analytical columns (CIMac) depletion column with specificity to HSA (CIMac-αHSA). Because of the convective flow-through channels, the polymethacrylate CIMac monoliths afford flow rate independent binding capacity and resolution that results in relatively short analysis time compared with traditional chromatographic supports. Seppro IgY14 depletion kit was used as a benchmark to control the results of depletion. Bottom-up proteomic approach followed by label-free quantitation using normalized spectral indexes were employed for protein quantification in G1/G2 and cleavage/blastocyst in vitro fertilization culture media widely utilized in clinics for embryo growth in vitro. The results revealed approximately equal HSA level of 100 ± 25% in albumin-enriched fractions relative to the nondepleted samples for both CIMac-αHSA column and Seppro kit. In the albumin-free fractions concentrated 5.5-fold by volume, serum albumin was identified at the levels of 5-30% and 20-30% for the CIMac-αHSA and Seppro IgY14 spin columns, respectively. PMID:27122488

  20. Depletion of human serum albumin in embryo culture media for in vitro fertilization using monolithic columns with immobilized antibodies.

    PubMed

    Tarasova, Irina A; Lobas, Anna A; Černigoj, Urh; Solovyeva, Elizaveta M; Mahlberg, Barbara; Ivanov, Mark V; Panić-Janković, Tanja; Nagy, Zoltan; Pridatchenko, Marina L; Pungor, Andras; Nemec, Blaž; Vidic, Urška; Gašperšič, Jernej; Krajnc, Nika Lendero; Vidič, Jana; Gorshkov, Mikhail V; Mitulović, Goran

    2016-09-01

    Affinity depletion of abundant proteins such as HSA is an important stage in routine sample preparation prior to MS/MS analysis of biological samples with high range of concentrations. Due to the charge competition effects in electrospray ion source that results in discrimination of the low-abundance species, as well as limited dynamic range of MS/MS, restricted typically by three orders of magnitude, the identification of low-abundance proteins becomes a challenge unless the sample is depleted from high-concentration compounds. This dictates a need for developing efficient separation technologies allowing fast and automated protein depletion. In this study, we performed evaluation of a novel immunoaffinity-based Convective Interaction Media analytical columns (CIMac) depletion column with specificity to HSA (CIMac-αHSA). Because of the convective flow-through channels, the polymethacrylate CIMac monoliths afford flow rate independent binding capacity and resolution that results in relatively short analysis time compared with traditional chromatographic supports. Seppro IgY14 depletion kit was used as a benchmark to control the results of depletion. Bottom-up proteomic approach followed by label-free quantitation using normalized spectral indexes were employed for protein quantification in G1/G2 and cleavage/blastocyst in vitro fertilization culture media widely utilized in clinics for embryo growth in vitro. The results revealed approximately equal HSA level of 100 ± 25% in albumin-enriched fractions relative to the nondepleted samples for both CIMac-αHSA column and Seppro kit. In the albumin-free fractions concentrated 5.5-fold by volume, serum albumin was identified at the levels of 5-30% and 20-30% for the CIMac-αHSA and Seppro IgY14 spin columns, respectively.

  1. Sperm apoptosis in nonpregnant luteal phase sera after in vitro fertilization as assessed by comparative genomic hybridization.

    PubMed

    Bouma, C L; Patton, W C; Jacobson, J D; King, A; Chan, P J

    2004-01-01

    Toxicity in serum has been reported in cases of recurrent spontaneous abortions and endometriosis. The null hypothesis was that serum toxicity was not involved in failed pregnancies after in vitro fertilization procedures. The objective was to expose donor sperm to pregnant versus nonpregnant patient sera and analyze for sperm DNA damaging effects using a novel comparative genomic hybridization method. Luteal phase sera (N = 21 cases) were drawn one week after embryo transfer. Colloid-washed donor sperm were incubated (48 h, 37 degrees C, 5% CO2 in air) in 0% or 50% sera. Single-stranded DNA (ssDNA) of control sperm were stained in Hoechst 33342 and hybridized to Sybr Gold-stained ssDNA of sera-treated sperm. Image analyses were performed and fluorescent intensities analyzed. Nonpregnant patient sera (57% of cases) were associated with DNA fragmentation (64.4 +/- 8.8 pixels; mean +/- S.E.M.) when compared with pregnant patient sera (106.3 +/- 8.4 pixels). There were no differences in the sera of biochemical (108.2 +/- 15.3) versus clinical pregnancy cases (105.3 +/- 11.4). The results suggest that nonpregnant patient sera contained factor(s) that cause DNA fragmentation leading to pregnancy losses.

  2. Predictive value of early serum beta-human chorionic gonadotrophin for the successful outcome in women undergoing in vitro fertilization

    PubMed Central

    Singh, Neeta; Goyal, Manu; Malhotra, Neena; Tiwari, Abanish; Badiger, Shreenivas

    2013-01-01

    AIMS: Pregnancies achieved by in vitro fertilization (IVF) are at increased risk of adverse outcome. The main objective of this study was to evaluate the predictive value of β-human chorionic gonadotrophin (β-HCG) and age of the patient for the successful outcome in IVF. MATERIALS AND METHODS: A retrospective study was done in 139 pregnancies after IVF at single IVF center from June 2007 to July 2012. The age of the patient and initial serum values of β-HCG on day 14 of embryo transfer were correlated with ongoing pregnancy (>12 weeks gestation). RESULTS: The β-HCG level on day 14 of more than 347 mIU/ml has a sensitivity of 72.2% and specificity of 73.6% in prediction of pregnancy beyond 12 weeks period of gestation. Positive likelihood ratio (LR) is 2.74 and negative LR is 0.37, (receiver operating characteristic area = 0.79). DISCUSSION: In IVF cycles, there is a lot of stress on the couples while the cycle is going on. There was a positive correlation between the higher values of early serum β-HCG levels and ongoing pregnancy. Hence, it can be used as an independent predictor of a successful outcome of IVF cycle. CONCLUSION: We concluded from our study that early serum β-HCG can be used as a predictor of a successful outcome in IVF. PMID:24672163

  3. Differences in body image between anorexics and in-vitro-fertilization patients - a study with Body Grid

    PubMed Central

    Borkenhagen, Ada; Klapp, Burghard F.; Schoeneich, Frank; Brähler, Elmar

    2005-01-01

    Objectives: The purpose of the investigation was to explore the body image disturbance of anorexics and in-vitro-fertilization patients (IvF-patients) with Body Grid and Body Identity Plot. Methods: The paper reports on an empirical study conducted with 32 anorexic patients and 30 IvF-patients. The structure of the body image was derived from the Body Grid, an idiographic approach following the Role Repertory Grid developed by George A. Kelly [17]. The representation of the body image and the degree of body-acceptance is represented graphically. Results: By the Body Grid and Body Identity Plot measures we were able to identify important differences in body image between anorexics and IvF-patients. Conclusion: The tendencies of dissociation in the body image of anorexics which we found must be seen in the sense of a specific body image disturbance which differs significantly from the body-experience profile of IvF-patients. With the grid approach it was possible to elicit the inner structure of body image and determine the acceptance of the body and integration of single body parts. PMID:19742059

  4. Pronuclear morphology evaluation in in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI) cycles: a retrospective clinical review

    PubMed Central

    2013-01-01

    Background The assessment of the embryo quality is crucial to maintain an high pregnancy rate and to reduce the risk of multiple pregnancy. The evaluation of the pronuclear and nucleolar characteristics of human zygote have been proposed as an indicator of embryo development and chromosomal complement. The aim of the current study was to assess the role of pronuclear morphology evaluation in vitro fertilization (IVF) / intracytoplasmic sperm injection (ICSI) cycles. Methods Retrospective clinical analysis on 755 non-elective transfers of only one embryo (ET). Embryo assessment was performed in days 1 and 2. Clinical and biological data were recorded and analyzed according to embryo and/or pronuclear morphology. Results Both pronuclear and embryo morphology were significantly related to clinical pregnancy and live-birth rates. No significant difference in clinical pregnancy and live-birth rates was detected when the pronuclear and embryo morphology assessments were combined. Embryo morphology and maternal age were the only independent predictors of favorable outcome by logistic regression analysis. Conclusions Pronuclear evaluation is effective to select the best zygotes if ET is performed at day 1, whereas it did not improve the clinical outcomes when combined with embryo morphology evaluation in day 2. PMID:23282023

  5. Pregnancies established from water buffalo (Bubalus bubalis) blastocysts derived from in vitro matured, in vitro fertilized oocytes and co-cultured with cumulus and oviductal cells.

    PubMed

    Madan, M L; Chauhan, M S; Singla, S K; Manik, R S

    1994-09-01

    Buffalo ovaries were collected immediately after slaughter and were transported to laboratory in sterile saline at 37 degrees C. Follicular oocytes with the cumulus mass aspirated from 2 to 6 mm in diameter follicles were cultured in TCM-199 medium supplemented with 10% buffalo estrus serum (BES) in 5% CO(2) at 38.5 degrees C. After 20 to 24 h of incubation, the oocytes were inseminated with precapacitated frozen thawed spermatozoa for 6 h. The fertilization rate was 78.15% of the matured oocytes. Over an in vitro culture period of 3 to 9 d, 4.02% of the inseminated oocytes developed to the morula stage when cultured with cumulus cells alone and 17.83% when cumulus cells plus oviductal epithelial cells were used. The percentage of developed blastocysts was very low (0.57%) when the oocytes were co-cultured with cumulus cells from the original oocytes. However, 8% of the inseminated oocytes that were denuded 3 d after insemination developed to the blastocyst stage when they were co-cultured with cumulus and oviductal epithelial cells. Sixteen early/expanded blastocysts were transferred non-surgically to 16 recipients. Four of the 16 recipients became pregnant, of which 2 delivered normal buffalo male calves.

  6. Ca2+ ionophore A23187 can make mouse spermatozoa capable of fertilizing in vitro without activation of cAMP-dependent phosphorylation pathways

    PubMed Central

    Tateno, Hiroyuki; Krapf, Dario; Hino, Toshiaki; Sánchez-Cárdenas, Claudia; Darszon, Alberto; Yanagimachi, Ryuzo; Visconti, Pablo E.

    2013-01-01

    Ca2+ ionophore A23187 is known to induce the acrosome reaction of mammalian spermatozoa, but it also quickly immobilizes them. Although mouse spermatozoa were immobilized by this ionophore, they initiated vigorous motility (hyperactivation) soon after this reagent was washed away by centrifugation. About half of live spermatozoa were acrosome-reacted at the end of 10 min of ionophore treatment; fertilization of cumulus-intact oocytes began as soon as spermatozoa recovered their motility and before the increase in protein tyrosine phosphorylation, which started 30–45 min after washing out the ionophore. When spermatozoa were treated with A23187, more than 95% of oocytes were fertilized in the constant presence of the protein kinase A inhibitor, H89. Ionophore-treated spermatozoa also fertilized 80% of oocytes, even in the absence of HCO3−, a component essential for cAMP synthesis under normal in vitro conditions. Under these conditions, fertilized oocytes developed into normal offspring. These data indicate that mouse spermatozoa treated with ionophore are able to fertilize without activation of the cAMP/PKA signaling pathway. Furthermore, they suggest that the cAMP/PKA pathway is upstream of an intracellular Ca2+ increase required for the acrosome reaction and hyperactivation of spermatozoa under normal in vitro conditions. PMID:24128762

  7. Developmental competence of different quality bovine oocytes retrieved through ovum pick-up following in vitro maturation and fertilization.

    PubMed

    Saini, N; Singh, M K; Shah, S M; Singh, K P; Kaushik, R; Manik, R S; Singla, S K; Palta, P; Chauhan, M S

    2015-12-01

    In the present study, oocytes retrieved from cross bred Karan Fries cows by ovum pick-up technique were graded into Group 1 and Group 2, based on the morphological appearance of the individual cumulus-oocyte complexes (COCs). To analyze whether the developmental potential of the COCs bears a relation to morphological appearance, relative expression of a panel of genes associated with; (a) cumulus-oocyte interaction (Cx43, Cx37, GDF9 and BMP15), (b) fertilization (ZP2 and ZP3), (c) embryonic development (HSF1, ZAR1 and bFGF) and (d) apoptosis and survival (BAX, BID and BCL-XL, MCL-1, respectively) was studied at two stages: germinal vesicle (GV) stage and after in vitro maturation. The competence was further corroborated by evaluating the embryonic progression of the presumed zygotes obtained from fertilization of the graded COCs. The gene expression profile and development rate in pooled A and B grade (Group 1) COCs and pooled C and D grade (Group 2) COCs were determined and compared according to the original grades. The results of the study demonstrated that the morphologically characterized Group 2 COCs showed significantly (P<0.05) lower expression for most of the genes related to cumulus-oocyte interplay, fertilization and embryonic development, both at GV stage as well as after maturation. Group 1 COCs also showed greater expression of anti-apoptotic genes (BCL-XL and MCL1) both at GV stage and after maturation, while pro-apoptotic genes (BAX and BID) showed significantly (P<0.05) elevated expression in poor quality COCs at both the stages. The cleavage rate in Group 1 COCs was significantly higher than that of Group 2 (74.46±7.06 v. 31.57±5.32%). The development of the presumed zygotes in Group 2 oocytes proceeded up to 8- to 16-cell stages only, while in Group 1 it progressed up to morulae (35.38±7.11%) and blastocyst stages (9.70±3.15%), indicating their better developmental potential.

  8. Urinary Phthalate Metabolite Concentrations and Reproductive Outcomes among Women Undergoing in Vitro Fertilization: Results from the EARTH Study

    PubMed Central

    Hauser, Russ; Gaskins, Audrey J.; Souter, Irene; Smith, Kristen W.; Dodge, Laura E.; Ehrlich, Shelley; Meeker, John D.; Calafat, Antonia M.; Williams, Paige L.

    2015-01-01

    Background: Evidence from both animal and human studies suggests that exposure to phthalates may be associated with adverse female reproductive outcomes. Objective: We evaluated the associations between urinary concentrations of phthalate metabolites and outcomes of assisted reproductive technologies (ART). Methods: This analysis included 256 women enrolled in the Environment and Reproductive Health (EARTH) prospective cohort study (2004–2012) who provided one to two urine samples per cycle before oocyte retrieval. We measured 11 urinary phthalate metabolites [mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), mono-isobutyl phthalate (MiBP), mono-n-butyl phthalate (MBP), monobenzyl phthalate (MBzP), monoethyl phthalate (MEP), monocarboxyisooctyl phthalate (MCOP), monocarboxyisononyl phthalate (MCNP), and mono(3-carboxypropyl) phthalate (MCPP)]. We used generalized linear mixed models to evaluate the association of urinary phthalate metabolites with in vitro fertilization (IVF) outcomes, accounting for multiple IVF cycles per woman. Results: In multivariate models, women in the highest as compared with lowest quartile of MEHP, MEHHP, MEOHP, MECPP, ΣDEHP (MEHP + MEHHP + MEOHP + MECPP), and MCNP had lower oocyte yield. Similarly, the number of mature (MII) oocytes retrieved was lower in the highest versus lowest quartile for these same phthalate metabolites. The adjusted differences (95% CI) in proportion of cycles resulting in clinical pregnancy and live birth between women in the fourth versus first quartile of ΣDEHP were –0.19 (–0.29, –0.08) and –0.19 (–0.28, –0.08), respectively, and there was also a lower proportion of cycles resulting in clinical pregnancy and live birth for individual DEHP metabolites. Conclusions: Urinary concentrations of DEHP metabolites were inversely associated with oocyte yield, clinical pregnancy

  9. The Effects of Di-(2-ethylhexyl)-phthalate Exposure on Fertilization and Embryonic Development In Vitro and Testicular Genomic Mutation In Vivo

    PubMed Central

    Gu, Yi-Hua; Liu, Miao; Xu, Yan; Yuan, Yao; Sun, Fei; Zhang, Hui-Qin; Shi, Hui-Juan

    2012-01-01

    The present study was undertaken to determine the reproductive hazards of Di-(2-ethylhexyl)-phthalate (DEHP) on mouse spermatozoa and embryos in vitro and genomic changes in vivo. Direct low-level DEHP exposure (1 μg/ml) on spermatozoa and embryos was investigated by in vitro fertilization (IVF) process, culture of preimplanted embryos in DEHP-supplemented medium and embryo transfer to achieve full term development. Big Blue® transgenic mouse model was employed to evaluate the mutagenesis of testicular genome with in vivo exposure concentration of DEHP (500 mg/kg/day). Generally, DEHP-treated spermatozoa (1 μg/ml, 30 min) presented reduced fertilization ability (P<0.05) and the resultant embryos had decreased developmental potential compared to DMSO controls (P<0.05). Meanwhile, the transferred 2-cell stage embryos derived from treated spermatozoa also exhibited decreased birth rate than that of control (P<0.05). When fertilized oocytes or 2-cell stage embryos were recovered by in vivo fertilization (without treatment) and then exposed to DEHP, the subsequent development proceed to blastocysts was different, fertilized oocytes were significantly affected (P<0.05) whereas developmental progression of 2-cell stage embryos was similar to controls (P>0.05). Testes of the Big Blue® transgenic mice treated with DEHP for 4 weeks indicated an approximately 3-fold increase in genomic DNA mutation frequency compared with controls (P<0.05). These findings unveiled the hazardous effects of direct low-level exposure of DEHP on spermatozoa's fertilization ability as well as embryonic development, and proved that in vivo DEHP exposure posed mutagenic risks in the reproductive organ – at least in testes, are of great concern to human male reproductive health. PMID:23226291

  10. The effects of Di-(2-ethylhexyl)-phthalate exposure on fertilization and embryonic development in vitro and testicular genomic mutation in vivo.

    PubMed

    Huang, Xue-Feng; Li, Yan; Gu, Yi-Hua; Liu, Miao; Xu, Yan; Yuan, Yao; Sun, Fei; Zhang, Hui-Qin; Shi, Hui-Juan

    2012-01-01

    The present study was undertaken to determine the reproductive hazards of Di-(2-ethylhexyl)-phthalate (DEHP) on mouse spermatozoa and embryos in vitro and genomic changes in vivo. Direct low-level DEHP exposure (1 μg/ml) on spermatozoa and embryos was investigated by in vitro fertilization (IVF) process, culture of preimplanted embryos in DEHP-supplemented medium and embryo transfer to achieve full term development. Big Blue® transgenic mouse model was employed to evaluate the mutagenesis of testicular genome with in vivo exposure concentration of DEHP (500 mg/kg/day). Generally, DEHP-treated spermatozoa (1 μg/ml, 30 min) presented reduced fertilization ability (P<0.05) and the resultant embryos had decreased developmental potential compared to DMSO controls (P<0.05). Meanwhile, the transferred 2-cell stage embryos derived from treated spermatozoa also exhibited decreased birth rate than that of control (P<0.05). When fertilized oocytes or 2-cell stage embryos were recovered by in vivo fertilization (without treatment) and then exposed to DEHP, the subsequent development proceed to blastocysts was different, fertilized oocytes were significantly affected (P<0.05) whereas developmental progression of 2-cell stage embryos was similar to controls (P>0.05). Testes of the Big Blue® transgenic mice treated with DEHP for 4 weeks indicated an approximately 3-fold increase in genomic DNA mutation frequency compared with controls (P<0.05). These findings unveiled the hazardous effects of direct low-level exposure of DEHP on spermatozoa's fertilization ability as well as embryonic development, and proved that in vivo DEHP exposure posed mutagenic risks in the reproductive organ - at least in testes, are of great concern to human male reproductive health.

  11. Predictive value of sperm morphology and movement characteristics in the outcome of in vitro fertilization of human oocytes.

    PubMed

    Chan, S Y; Wang, C; Chan, S T; Ho, P C; So, W W; Chan, Y F; Ma, H K

    1989-06-01

    One hundred fourteen semen samples from Chinese males were analyzed for routine semen parameters including the semen volume, sperm count, percentage motility, and percentage normal morphology. Of these 114 samples, 54 also had movement characteristics of seminal and swim-up sperm evaluated by the computer image analyzer system (Cellsoft; Cryo Resources Co., New York). All semen samples were subjected to the swim-up procedure to harvest the motile sperm before inseminations of human oocytes. Fertilization was considered to have occurred when at least one oocyte was observed with two or more pronuclei. Semen samples were classified as infertile (0% fertilization rate; N = 32) or fertile (greater than 0% fertilization rate; N = 82) before statistical analyses. There was a significant difference (P less than 0.005) in percentage normal morphology of seminal sperm between the fertile (mean +/- SE; 67.3 +/- 1.2%) and the infertile (59.3 +/- 2.2%) samples. The percentage normal morphology of seminal sperm correlated (r = 0.3049; P less than 0.002) with the fertilization rate and this parameter was selected by the multivariate stepwise discriminant analysis as the discriminator capable of predicting the fertilization rate with 57.9% accuracy. Statistical analyses of samples where sperm movement was also evaluated demonstrated that there was significant differences (P less than 0.01) between the fertile (N = 38) and the infertile (N = 16) samples in percentage normal morphology of seminal sperm (67.8 +/- 1.8% vs 56.2 +/- 2.6%) and curvilinear velocity of swim-up sperm (89.2 +/- 3.5 vs 68.2 +/- 7.2 microns/sec).(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Singleton Pregnancy Outcomes after In Vitro Fertilization with Fresh or Frozen-Thawed Embryo Transfer and Incidence of Placenta Praevia

    PubMed Central

    Korosec, Sara; Kladnik, Urska; Kotar, Vanja; Virant-Klun, Irma; Vrtacnik Bokal, Eda

    2014-01-01

    The aim of the study was to compare the single pregnancy and neonate outcome after fresh and frozen-thawed embryo transfer in the in vitro fertilization programme (IVF). The study focused on clinical and laboratory factors affecting the abnormal placentation, especially placenta praevia, in patients conceiving in the IVF programme. The results confirm that neonates born after frozen-thawed embryo transfer had significantly higher mean birth weight than after fresh embryo transfer (ET). Moreover, the birth weight distribution in singletons was found to shift towards “large for gestation” (LGA) after frozen-thawed ET. On the other hand, the pregnancies after fresh ET were characterized by a higher incidence of placenta praevia and 3rd trimester bleeding. Placenta praevia was more common in IVF patients with fresh ET in a stimulated cycle than in patients with ET in a spontaneous cycle. It occurred more frequently in patients with transfer of 2 embryos. From this point of view, single ET and ET in a spontaneous cycle should be encouraged in good prognosis patients in the future with more than two good quality embryos developed. An important issue arose of how the ovarian hormonal stimulation relates to abnormal placentation and if the serum hormone levels interfere with in the IVF treatment results. PMID:24822209

  13. Assessment of pronuclear formation following in vitro fertilization with bovine spermatozoa obtained after thermal insulation of the testis.

    PubMed

    Walters, A H; Saacke, R G; Pearson, R E; Gwazdauskas, F C

    2006-04-01

    This study was conducted to follow the chronology of pronuclear formation in bovine zygotes after in vitro insemination with a population of spermatozoa having abnormal morphology. Semen samples were obtained and cryopreserved from four Holstein bulls before and after a scrotal insulation period of 48 h (Day 0). A pre-insult (Day 5) and a Day 20 post-insult semen sample were evaluated for morphology and used for IVF after standard swim-up sperm separation protocols. Pronuclear formation was scored on subpopulations of presumptive zygotes after they were fixed and stained at 3-h time intervals from 6 to 18 h post in vitro insemination (hpi). Post-thaw morphological evaluation of semen samples revealed a decrease in the percentages of normal spermatozoa in the post-insult samples compared with the pre-insult samples for Bull I (74-22%) and Bull III (68-1%). The sperm penetration rate decreased (P<0.05) between the pre- and post-insult samples for Bulls I (90-76%) and III (92-70%), but was not different for Bulls II (92-90%) and IV (78-85%). The pronuclear formation rates for post-insult zygotes for Bulls II and IV had comparable increases in development over time, whereas there was no increase in the pronuclear development for the zygotes from the post-insult samples for Bulls I and III, and generally a condensed sperm head was observed in the oolemma. At 18 hpi the fertilization rate between the pre- and post-insult samples for Bulls I (51-4%), II (88-75%) and III (94-2%) decreased (P<0.01), but there was no change for Bull IV (66%). In conclusion, we inferred that the failure in normal pronuclear formation was associated with an absence of normal decondensation of the penetrating spermatozoon; this suggested that the effect of morphologically abnormal spermatozoa occurred prior to cleavage, thus limiting early development.

  14. Characterization of pig sperm hyaluronidase and improvement of the digestibility of cumulus cell mass by recombinant pSPAM1 hyaluronidase in an in vitro fertilization assay.

    PubMed

    Yoon, Sungwon; Chang, Kyu-Tae; Cho, Hongsang; Moon, Jisang; Kim, Ju-Sung; Min, Sung-Hun; Koo, Deog-Bon; Lee, Sang-Rae; Kim, Sang-Hyun; Park, Ki-Eun; Park, Young Il; Kim, Ekyune

    2014-11-30

    Although sperm hyaluronidase is thought to play an important role in mammalian fertilization, the molecular function underlying these steps remains largely unknown. In mouse models, sperm-specific SPAM1 and HYAL5 hyaluronidase are believed to function in both sperm penetration of the cumulus matrix and sperm-ZP binding. However, gene-targeting studies for SPAM1 or HYAL5 show that hyaluronidases are not essential for fertilization, despite the fact that exogenous hyaluronidase can disrupt the cumulus matrix. Therefore, to evaluate whether sperm hyaluronidase is essential for mammalian fertilization, it is necessary to generate HYAL5/SPAM1 double-knockout mice. However, generating double-knockout mice is very difficult because these two genes exist on the same chromosome. Recently, investigators have begun to employ the pig model system to study human disease due to its similarities to human anatomy and physiology. In this study, we confirmed that pig SPAM1 exists as a single copy gene on chromosome 18 and is specifically expressed in the testis. In addition, we expressed recombinant pig SPAM1 in human embryonic kidney 293 cells and showed that these enzymes possess hyaluronidase activity. We also demonstrated that a polyclonal antibody against pig sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggest that pig SPAM1 may play a critical role in pig fertilization and that recombinant SPAM1 can disperse the oocyte-cumulus complex in an IVF assay.

  15. Debating Elective Single Embryo Transfer after in vitro Fertilization: A Plea for a Context-Sensitive Approach

    PubMed Central

    Ezugwu, EC; Van der Burg, S

    2015-01-01

    The number of embryos transferred after in vitro fertilization (IVF) have been a topic of debate for over a decade now. Due to the risk associated with multiple pregnancy, there has been a global effort at reducing the multiple pregnancy rates to a minimum while maintaining an acceptable level of successful IVF pregnancy rate. Elective single embryo transfer (eSET) is advocated in most European countries. In Belgium and Sweden, eSET is mandatory for couples with a good prognosis. However, despite clinical recommendations and policy statements, patients in clinical practice frequently do request for the transfer of multiple embryos in order to have twins. Such requests conflict with policy guidelines and create an ethical dilemma for physicians: Should the physician do as the couple requests, and there with respect the autonomy of patients, or adhere to medical policy that takes the health of the mother and children at heart? This article provides an exploration of the arguments found in the literature that plays a role in the discussion on this topic and eventually argues that what a physician should do depends on the specificities of the context in which patients and physicians are implicated. These contextual issues can be taken into account in a shared decision-making procedure, which allows reflections and the responsibilities of both patients and physicians to be attended in decision about assisted reproduction. PMID:25745568

  16. The Impact of Serum Progesterone Levels on the Results of In Vitro Fertilization Treatments: A Literature Review.

    PubMed

    Castillo, Jaime Larach Del; Bousamra, Maroun; Fuente, Laura De La; Ruiz-Balda, Jose A; Palomo, Marissa

    2015-08-01

    The aim of this review is to analyze the relationship between preovulatory progesterone (P) rise and in vitro fertilization (IVF) pregnancy outcomes. It also investigates the sources and effects of rises in progesterone levels, including the underlying mechanisms and potential strategies in preventing its elevation during ovarian stimulation. Progesterone is produced in the early follicular phase in the adrenal gland, which shifts toward the ovaries prior to ovulation. Several factors contribute to the etiology of P level increase including the number of multiple follicles, the overdose of gonadotropins and poor ovarian response. Nowadays, the influence of the preovulatory P rise on IVF outcome remains controversial. Several authors have failed to demonstrate any negative impact, while others reported a detrimental effect associated with the rise of P. It seems that P rise (1.5 ng/ml or 4.77 nmol/l) may have deleterious effects on endometrial receptivity, namely, accelerating the endometrial maturation process that subsequently narrows the period for implantation and thus decreases pregnancy rates. Recent studies have proposed different cutoffs according to the ovarian response, which may be a little high in patients with high response in relation to those of normal response or low response. To prevent a P rise, it might be preferable to use milder stimulation protocols, earlier trigger of ovulation, cryopreservation of all embryos and transfer in the natural cycle.

  17. Pronuclear morphology evaluation for fresh in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles: a systematic review

    PubMed Central

    2013-01-01

    The current systematic review was aimed to assess the effectiveness of the zygote morphology evaluation in fresh in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles. All available studies reporting on zygote morphology and clinical and/or biological outcomes were analyzed. Forty studies were included in the final analysis. Fourteen different zygote scoring systems were employed. Zygote morphology correlated significantly with embryo quality and cleavage, blastocyst stage, embryonic chromosome status, in a high proportion of the studies which assessed the specific outcome [15/25 (60%), 15/20 (75%), 7/8 (87.5%), 6/6 (100%), respectively]. On the other hand, only a reduced proportion of papers showed a statistically significant relationship between implantation, pregnancy and delivery/live-birth rates and zygote morphology score [12/23 (52.2%), 12/25 (48%), 1/4 (25%), respectively]. In conclusion, our findings demonstrate the lack of conclusive data on the clinical efficacy of the zygote morphology evaluation in fresh IVF/ICSI cycles, even if biological results showing a good relationship with embryo viability suggest a role in cycles in which the transfer/freezing is performed at day 1. PMID:24028277

  18. The Impact of Serum Progesterone Levels on the Results of In Vitro Fertilization Treatments: A Literature Review.

    PubMed

    Castillo, Jaime Larach Del; Bousamra, Maroun; Fuente, Laura De La; Ruiz-Balda, Jose A; Palomo, Marissa

    2015-01-01

    The aim of this review is to analyze the relationship between preovulatory progesterone (P) rise and in vitro fertilization (IVF) pregnancy outcomes. It also investigates the sources and effects of rises in progesterone levels, including the underlying mechanisms and potential strategies in preventing its elevation during ovarian stimulation. Progesterone is produced in the early follicular phase in the adrenal gland, which shifts toward the ovaries prior to ovulation. Several factors contribute to the etiology of P level increase including the number of multiple follicles, the overdose of gonadotropins and poor ovarian response. Nowadays, the influence of the preovulatory P rise on IVF outcome remains controversial. Several authors have failed to demonstrate any negative impact, while others reported a detrimental effect associated with the rise of P. It seems that P rise (1.5 ng/ml or 4.77 nmol/l) may have deleterious effects on endometrial receptivity, namely, accelerating the endometrial maturation process that subsequently narrows the period for implantation and thus decreases pregnancy rates. Recent studies have proposed different cutoffs according to the ovarian response, which may be a little high in patients with high response in relation to those of normal response or low response. To prevent a P rise, it might be preferable to use milder stimulation protocols, earlier trigger of ovulation, cryopreservation of all embryos and transfer in the natural cycle. PMID:27203093

  19. Blood plasma collected after adrenocorticotropic hormone administration during the preovulatory period in the sow negatively affects in vitro fertilization by disturbing spermatozoa function.

    PubMed

    González, R; Kumaresan, A; Bergqvist, A S; Sjunnesson, Y C B

    2015-04-15

    Successful fertilization is essential for reproduction and might be negatively affected by stressful events, which could alter the environment where fertilization occurs. The aim of the study was to determine whether an altered hormonal profile in blood plasma caused by adrenocorticotropic hormone (ACTH) administration could affect in vitro fertilization in the pig model. In experiment 1, gametes were exposed for 24 hours to plasma from ACTH-treated, non-ACTH-treated sows, or medium with BSA. Fertilization, cleavage, and blastocyst rates were lower in the ACTH group compared with the no ACTH or BSA control groups (P < 0.01). In experiment 2, the exposure of matured oocytes for 1 hour before fertilization to the same treatments did not have an impact on their ability to undergo fertilization or on embryo development. In experiment 3, spermatozoa were incubated for 0, 1, 4, and 24 hours under the same conditions. There was no effect of treatment on sperm viability. The percentage of acrosome-reacted spermatozoa remained higher in the ACTH group compared with the non-ACTH-treated group through the incubation period (P < 0.001). Protein tyrosine phosphorylation (PTP) patterns were also affected by treatment (P < 0.001). The presence of an atypical PTP pattern was higher in the ACTH group at all the analyzed time points compared with the BSA and no ACTH groups (P < 0.001). In conclusion, this altered environment may not affect oocyte competence but might affect the sperm fertilizing ability through alterations in the acrosome reaction and correct sequence of PTP patterns.

  20. Endometrial fluid associated with Essure implants placed before in vitro fertilization: Considerations for patient counseling and surgical management

    PubMed Central

    Walsh, David J; Jones, Christopher A; Wood, Samuel H

    2015-01-01

    Essure (Bayer) received approval from the U.S. Food and Drugs Administration as a permanent non-hormonal contraceptive implant in November 2002. While the use of Essure in the management of hydrosalpinx prior to in vitro fertilization (IVF) remains off-label, it has been used specifically for this purpose since at least 2007. Although most published reports on Essure placement before IVF have been reassuring, clinical experience remains limited, and no randomized studies have demonstrated the safety or efficacy of Essure in this context. In fact, no published guidelines deal with patient selection or counseling regarding the Essure procedure specifically in the context of IVF. Although Essure is an irreversible birth control option, some patients request the surgical removal of the implants for various reasons. While these patients could eventually undergo hysterectomy, at present no standardized technique exists for simple Essure removal with conservation of the uterus. This article emphasizes new aspects of the Essure procedure, as we describe the first known association between the placement of Essure implants and the subsequent development of fluid within the uterine cavity, which resolved after the surgical removal of both devices. PMID:26473113

  1. The Experience of Chinese Couples Undergoing In Vitro Fertilization Treatment: Perception of the Treatment Process and Partner Support

    PubMed Central

    Ying, Li-Ying; Wu, Lai Har; Loke, Alice Yuen

    2015-01-01

    Background Couples undergoing In Vitro Fertilization (IVF) Treatment suffer as dyads from the stressful experience of the painful treatment and the fear that the IVF cycle will fail. They are likely to report that their marital relationship has become unstable due to the prolonged period of treatment. Methods This is a qualitative study that was conducted to explore the experiences that Chinese couples have had with IVF treatment, especially their perceptions of the process and the support between couples. Results The interviews revealed that couples suffered from the process, experiencing physical and emotional pain, struggling with the urgency and inflexibility of bearing a child, and experiencing disturbances in their daily routines and work. The participants described how they endured the hardships as a couple and how it affected their relationship. The couples felt that sharing feelings and supporting each other contribute to psychological well-being and improves the marital relationship. They also identified some unfavorable aspects in their partner relationship. They were ambivalent about receiving social support from friends and family members. Conclusions With the couples indicating that the support that they received from each other affected their experience during the treatment process, it is suggested that a supportive intervention that focuses on enhancing the partnership of the couples and dealing with their inflexibility on the issue of bearing a child might result in improvements in the psychological status and marital relationship of infertile couples undergoing IVF treatment. PMID:26431545

  2. Good results of milder form of ovarian stimulation in an in vitro fertilization/intracytoplasmic sperm injection program.

    PubMed

    Grochowski, D; Wołczyński, S; Kuczyński, W; Domitrz, J; Szamatowicz, J; Szamatowicz, M

    1999-10-01

    In a prospective study, we compared two protocols of ovulation stimulation, the clomiphene citrate and human menopausal gonadotropin (hMG) versus D-triptorelin, a long-acting gonadotropin-releasing hormone (GnRH) agonist and hMG in 324 couples having their first in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) program, in terms of pregnancy rates and cost-effectiveness of drugs used. The GnRH agonist/hMG group was characterized by a greater mean number of ampoules of hMG used (31.7 versus 10.2), a larger number of oocytes collected (10.4 versus 4.2), and a larger number of embryos obtained (5.8 versus 2.9). With the policy of transferring only two of the best quality embryos, the mean number of embryos replaced were comparable (1.8 in clomiphene citrate/hMG and 1.9 in GnRH agonist/hMG group). The percentage of patients reaching embryo transfer was lower in the clomiphene citrate/hMG than in the GnRH agonist/hMG group (84.1% versus 93.1%, respectively). However, the combined results of the IVF and ICSI procedure in terms of pregnancy rate, both per patient and per embryo transfer were better, though not significantly in the clomiphene citrate/hMG than in GnRH agonist/hMG group (25.0% and 29.7% versus 23.7% and 25.5%, respectively). Similarly, the implantation rate was better (19.0% versus 13.5%, respectively). With the use of clomiphene citrate/hMG, a fivefold less costly drug regimen, we obtained pregnancy rates equivalent to those gained using GnRH agonist/hMG in our IVF/ICSI program.

  3. Effect of Laparoscopic Ovarian Drilling on Outcomes of In Vitro Fertilization in Clomiphene-Resistant Women with Polycystic Ovary Syndrome

    PubMed Central

    Eftekhar, Maryam; Deghani Firoozabadi, Razieh; Khani, Parisa; Ziaei Bideh, Ehsan; Forghani, Hosein

    2016-01-01

    Background Recently the laparoscopic ovarian drilling (LOD) has been used as a surgical treatment for ovulation in women with polycystic ovarian syndrome (PCOS), although its mechanism and outcomes are still unclear. This study was undertaken to evaluate the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) outcomes in clomiphene-resistant women with PCOS who were treated with LOD. Materials and Methods In this retrospective study, we reviewed the medical records of 300 women between 20 to 35 years old with clomiphene-resistant PCOS who had an ovulatory infertility and who were nominated for IVF/ICSI. Based on their treatment history, they were located into the following two groups: group I (n=150) including PCOS women who had history of LOD at least 6 months to 3 years before IVF/ICSI, and group II (n=150) including PCOS patients without history of drilling. Both groups were treated with antagonist protocol in the assisted reproductive technology (ART) process. The duration of treatment cycles, number of oocytes and embryos obtained, chemical and clinical pregnancy rate, the number of embryos transferred, and presence of ovarian hyper stimulation syndrome (OHSS) were measured. To compare means and frequencies, Student’s t test, Mann-whitney and chi-square tests were used. Results Our results showed that ovarian cauterization before IVF/ICSI in patients with PCOS reduced the risk of OHSS (P=0.025). Despite the same pregnancy rate in both groups (P=0.604), more obtained oocytes and embryos were seen on women without ovarian drilling than women with LOD (P˂0.001 and P=0.033, respectively). Conclusion There is no difference between the pregnancy rate in both groups. Due to significant reduction in OHSS in women undergoing LOD, this surgical treatment may be considered as a useful technique in the management of patients who have previously developed OHSS. However, there are ongoing concerns about long-term effects of LOD on ovarian function. PMID

  4. Addition of gonadotropin releasing hormone agonist for luteal phase support in in-vitro fertilization: an analysis of 2739 cycles

    PubMed Central

    Şimşek, Erhan; Kılıçdağ, Esra Bulgan; Aytaç, Pınar Çağlar; Çoban, Gonca; Şimşek, Seda Yüksel; Çok, Tayfun; Haydardedeoğlu, Bülent

    2015-01-01

    Objective Luteal phase is defective in in vitro fertilization (IVF) cycles, and many regimens were tried for the very best luteal phase support (LPS). Gonadotropin releasing hormone (GnRH) agonist use, which was administered as an adjunct to the luteal phase support in IVF cycles, was suggested to improve pregnancy outcome measures in certain randomized studies. We analyzed the effects of addition of GnRH agonist to standard progesterone luteal support on pregnancy outcome measures, particularly the live birth rates. Material and Methods This is a retrospective cohort study, including 2739 IVF cycles. Long GnRH agonist and antagonist stimulation IVF cycles with cleavage-stage embryo transfer were included. Cycles were divided into two groups: Group A included cycles with single-dose GnRH agonist plus progesterone LPS and Group B included progesterone only LPS. Live birth rates were the primary outcome measures of the analysis. Miscarriage rates and multiple pregnancy rates were the secondary outcome measures. Results Live birth rates were not statistically different in GnRH agonist plus progesterone (Group A) and progesterone only (Group B) groups in both the long agonist and antagonist stimulation arms (40.8%/41.2% and 32.8%/34.4%, p<0.05 respectively). Moreover, pregnancy rates, implantation rates, and miscarriage rates were found to be similar between groups. Multiple pregnancy rates in antagonist cycles were significantly higher in Group A than those in Group B (12.0% and 6.9%, respectively). Conclusion A beneficial effect of a single dose of GnRH agonist administration as a luteal phase supporting agent is yet to be determined because of the wide heterogeneity of data present in literature. Well-designed randomized clinical studies are required to clarify any effect of luteal GnRH agonist addition on pregnancy outcome measures with different doses, timing, and administration routes of GnRH agonists. PMID:26097392

  5. Increasing the cAMP concentration during in vitro maturation of pig oocytes improves cumulus maturation and subsequent fertilization in vitro.

    PubMed

    Appeltant, R; Beek, J; Vandenberghe, L; Maes, D; Van Soom, A

    2015-02-01

    Porcine IVF faces various problems such as incomplete cytoplasmic maturation of the oocyte and polyspermy. Previous studies proved the importance of cAMP in regulating nuclear and cytoplasmic maturation of oocytes. This study investigated the effect of the cAMP-modulating agents 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cAMP sodium salt (dbcAMP) on several parameters during in vitro production of porcine embryos. First, we wanted to see if oocyte collection in IBMX could meiotically arrest oocytes and, as such, improve synchronization of nuclear and cytoplasmic maturation. To this end, cumulus-oocyte complexes (COCs) were collected from gilts in HEPES-buffered Tyrode balanced salt solution medium with 0.5-mM IBMX or without IBMX. At the end of oocyte collection, the effect of IBMX on chromatin configuration was evaluated. However, no differences could be observed in nuclear configuration between IBMX- and IBMX+ oocytes (P > 0.05). Second, we added dbcAMP during IVM to improve cytoplasmic maturation and evaluated cumulus expansion (lack of adhesion), a disintegrin and metalloproteinase with thrombospondin-like repeats (ADAMTS-1) levels in cumulus cells, fertilization, and blastocyst rates. Cumulus-oocyte complexes were matured in modified North Carolina State University medium 37 with or without 1-mM dbcAMP. Frozen-thawed, epididymal, boar spermatozoa were used for IVF. After IVF, presumed zygotes were cultured for 7 days in North Carolina State University medium 23. Penetration rate decreased in dbcAMP+ (57.3%) compared with dbcAMP- (67.8%), but the polyspermy rate also decreased (43.3% vs. 53.4%, respectively) leading to an increased normal fertilization rate (56.7% vs. 46.6%, respectively; P < 0.05). Only 7.2% of the COCs showed adhesion in dbcAMP+ which was lower than 15.7% in dbcAMP- (P < 0.05) probably because of an upregulation of the ADAMTS-1 protein by dbcAMP. When the adherent oocytes were removed during maturation, no difference could be

  6. Pregnancy Outcomes of In Vitro Fertilization with or without Ovarian Hyperstimulation Syndrome: A Retrospective Cohort Study in Chinese Patients

    PubMed Central

    Jiang, Xuan; Deng, Cheng-Yan; Sun, Zheng-Yi; Chen, Wei-Lin; Wang, Han-Bi; Zhou, Yuan-Zheng; Jin, Li

    2015-01-01

    Background: The effect of ovarian hyperstimulation syndrome (OHSS) on pregnancy outcomes of in vitro fertilization (IVF) patients is still ambiguous. This study aimed to analyze pregnancy outcomes of IVF with or without OHSS in Chinese patients. Methods: A retrospective cohort study was undertaken to compare pregnancy outcomes between 190 women with OHSS and 197 women without OHSS. We examined the rates of clinical pregnancy, multiple pregnancies, miscarriage, live birth, preterm delivery, preterm birth before 34 weeks’ gestation, cesarean delivery, low birth weight (LBW), and small-for-gestational age (SGA) between the two groups. Odds ratios (ORs) and 95% confidence intervals (CIs) of measure of clinical pregnancy were also analyzed. Results: The clinical pregnancy rate of OHSS patients was significantly higher than that of non-OHSS patients (91.8% vs. 43.5%, P < 0.001). After controlling for drug protocol and causes of infertility, the adjusted ORs of moderate OHSS and severe/critical OHSS for clinical pregnancy were 4.65 (95% CI, 1.86–11.61) and 5.83 (95% CI, 3.45–9.86), respectively. There were no significant differences in rates of multiple pregnancy (4.0% vs. 3.7%) and miscarriage (16.1% vs. 17.5%) between the two groups. With regard to ongoing clinical pregnancy, we also found no significant differences in the rates of live birth (82.1% vs. 78.8%), preterm delivery (20.9% vs. 17.5%), preterm birth before 34 weeks’ gestation (8.6% vs. 7.9%), cesarean delivery (84.9% vs. 66.3%), LBW (30.2% vs. 23.5%), and SGA (21.9% vs. 17.6%) between the two groups. Conclusion: OHSS, which occurs in the luteal phase or early pregnancy in IVF patients and represents abnormal transient hemodynamics, does not exert any obviously adverse effect on the subsequent pregnancy. PMID:26612291

  7. Preimplantation Genetic Diagnosis (PGD) on In-Vitro Fertilization (IVF) Websites: Presentations of Risks, Benefits and Other Information

    PubMed Central

    Klitzman, Robert; Zolovska, Beata; Folberth, William; Sauer, Mark V.; Chung, Wendy; Appelbaum, Paul

    2010-01-01

    Objective To examine information on Preimplantation Genetic Diagnosis (PGD) presented on In-Vitro Fertilization (IVF) clinic websites. Design We systematically sampled every third IVF clinic on the 2004 CDC provider list. Setting The Internet. Patients None. Interventions None. Main Outcome Measures Benefits, risks and other types of information mentioned regarding PGD. Results Of 135 sites examined, 88.1% had websites, and 70% mentioned PGD, of which 27% were university/hospital-based and 63% were private clinics. Sites mentioning PGD listed uses/benefits of PGD far more than the risks involved. Of these sites, 76% described testing for single gene diseases, but fewer mentioned risks of missing target diagnoses (35%), or risks for loss of embryo (18%); and 14% described PGD as new or controversial. Private clinics were more likely than other programs to: be on either the East or West Coasts; list certain PGD risks (e.g., diagnostic error); note that PGD was new or controversial; reference source of PGD information; provide accuracy rates of genetic testing of embryos; and offer gender selection for social reasons. Conclusions Most IVF clinics advertise PGD on-line, but the scope and quality of information about it varies widely, emphasizing benefits while minimizing risks. Clinics and patients may benefit from more thorough and consistent presentation of PGD, drawing on available evidence to best provide a realistic portrayal of PGD. PMID:18829009

  8. Male mice produced by in vitro culture have reduced fertility and transmit organomegaly and glucose intolerance to their male offspring.

    PubMed

    Calle, Alexandra; Miranda, Alberto; Fernandez-Gonzalez, Raul; Pericuesta, Eva; Laguna, Ricardo; Gutierrez-Adan, Alfonso

    2012-08-01

    It has been reported that suboptimal in vitro culture (IVC) of mouse embryos can affect the postnatal expression of epigenetically sensitive alleles, resulting in altered postnatal growth, organ dimensions, health, and behavior in the offspring. Although these detrimental impacts on the offspring are well described, the relative contribution of the IVC-produced fathers is unclear. In this work, we have analyzed if suboptimal IVC (achieved by altering the culture medium by the addition of FCS) can affect male fertility and if organ size and glucose clearance, two of the adverse effects produced by suboptimal IVC conditions, were transmitted to the next two generations. IVC-produced males had lower sperm concentrations (5.8 × 10(6) spermatozoa in IVC vs. 14.5 × 10(6) spermatozoa in control), and these sperm exhibited decreased overall motility (49.6% vs. 72.8% in control) and progressive motility (22.6% vs. 32.2% in control). Fertility tests demonstrated that the percentage of pregnancies was reduced for IVC males (35% for IVC-produced males vs. 86% for in vivo controls). These features were related to a modified gene expression pattern in adult male testes, showing an altered gene expression in genes involved in DNA repair and apoptosis that was confirmed by TUNEL assay. Regarding the IVC related adverse phenotype transmitted to offspring, male glucose intolerance was shown only in F1 and F2 male but not female offspring. The same occurred with male abnormalities in the organ size of the liver, which were transmitted to F1 and F2 males but not to F1 females; moreover, analysis of the F0, F1, and F2 males revealed greater coefficients of variance in body weight and glucose intolerance than the control group. Finally, we analyzed, through gene silencing, the effect of IVC on the mRNA expression at the blastocyst stage for 11 known gene expression modifiers of epigenetic reprogramming. Suboptimal IVC reduced the expression of Kap1, Sox2, Hdac1, Dnmt1, and Dnmt3a

  9. Hormonal priming, induction of ovulation and in-vitro fertilization of the endangered Wyoming toad (Bufo baxteri)

    PubMed Central

    Browne, Robert K; Seratt, Jessica; Vance, Carrie; Kouba, Andrew

    2006-01-01

    The endangered Wyoming toad (Bufo baxteri) is the subject of an extensive captive breeding and reintroduction program. Wyoming toads in captivity rarely ovulate spontaneously and hormonal induction is used to ovulate females or to stimulate spermiation in males. With hormonal induction, ovulation is unreliable and egg numbers are low. The sequential administration of anovulatory doses of hormones (priming) has increased egg numbers and quality in both anurans and fish. Consequently, we tested the efficacy of a combination of human Chorionic Gonadotrophin (hCG) and Luteinizing Hormone Releasing Hormone analogue (LHRHa) administered as one dose, or two or three sequential doses to Bufo baxteri on egg numbers, fertilization and early embryo development. Spawning toads deposited eggs into Simplified Amphibian Ringers (SAR) solution to enable controlled in-vitro fertilization (IVF) with sperm from hormonally induced male toads. Unprimed females receiving a single mixed normally ovulatory dose of 500 IU hCG plus 4 micrograms of LHRHa produced no eggs. Whereas females primed with this dose and an anovulatory dose (100 IU hCG and 0.8 micrograms of LHRHa) of the same hormones, or primed only with an anovulatory dose, spawned after then receiving an ovulatory dose. Higher total egg numbers were produced with two primings than with one priming. Moreover, two primings produced significantly more eggs from each individual female than one priming. The cleavage rate of eggs was not found to differ between one or two primings. Nevertheless, embryo development with eggs from two primings gave a significantly greater percentage neurulation and swim-up than those from one priming. Of the male toads receiving a single dose of 300 IU hCG, 80% produced spermic urine with the greatest sperm concentration 7 hours post-administration (PA). However, peak sperm motility (95%) was achieved at 5 hours PA and remained relatively constant until declining 20 hours PA. In conclusion, Bufo baxteri

  10. Inhibitors of serine proteases decrease sperm penetration during porcine fertilization in vitro by inhibiting sperm binding to the zona pellucida and acrosome reaction.

    PubMed

    Beek, J; Nauwynck, H; Appeltant, R; Maes, D; Van Soom, A

    2015-11-01

    Serine proteases are involved in mammalian fertilization. Inhibitors of serine proteases can be applied to investigate at which point these enzymes exert their action. We selected two serine protease inhibitors, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF, 100 μM) and soybean trypsin inhibitor (STI, 5 μM) from Glycine max, via previous dose-response IVF experiments and sperm toxicity tests. In the present study, we evaluated how these inhibitors affect porcine fertilization in vitro as calculated on total fertilization rate, polyspermy rate, and the sperm number per fertilized oocyte of cumulus-intact, cumulus-free, and zona-free oocytes. In the control group (no inhibitor), these parameters were 86%, 49%, and 2.2 for cumulus-intact oocytes and 77%, 43%, and 2.2 for cumulus-free oocytes (6-hour gamete incubation period, 1.25 × 10(5) spermatozoa/mL). 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride and STI significantly reduced total fertilization and polyspermy rate in cumulus-intact and cumulus-free oocytes (P < 0.05). Total fertilization rates were respectively 65% and 53% (AEBSF) and 36% and 17% (STI). Inhibition rates were higher in cumulus-free oocytes than in cumulus-intact oocytes, indicating that inhibitors exerted their action after sperm passage through the cumulus. 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride but not STI reduced sperm binding to the ZP. The acrosome reaction was significantly inhibited by both inhibitors. Only 40.4% (AEBSF) and 11.4% (STI) of spermatozoa completed a calcium-induced acrosome reaction compared to 86.7% of spermatozoa in the control group. There was no effect on sperm binding or fertilization parameters in zona-free oocytes. In conclusion, sperm-zona binding and acrosome reaction were inhibited by serine protease inhibitors during porcine IVF.

  11. Oxidative stress biomarkers in endometrial secretions: A comparison between successful and unsuccessful in vitro fertilization cycles.

    PubMed

    Rahiminejad, Mohammad Ehsan; Moaddab, Amirhossein; Ganji, Maziar; Eskandari, Nika; Yepez, Mayel; Rabiee, Soghra; Wise, Meredith; Ruano, Rodrigo; Ranjbar, Akram

    2016-08-01

    A potential role of oxidative stress has been implicated in the outcome of various steps of assisted reproductive technology (ART). In a prospective cohort study, a total of 100 patients undergoing IVF/ICSI procedure due to male factor infertility were recruited based on the inclusion criteria. In all patients, 1-2ml of endometrial secretions was aspirated prior to embryo transfer. The oxidative stress markers in endometrial secretions, including superoxide dismutase (SOD), catalase (CAT) activities, lipid peroxidation (LPO), total thiol groups (TTG), and total antioxidant power (TAP) were investigated and compared among study groups including term pregnancy, failed IVF cycle, and miscarriage. P<0.05 was considered statistically different. Of the 100 patients, 28 cases (28%) resulted in ongoing pregnancy (biochemical pregnancy followed by clinical pregnancy), 11 cases (11%) resulted in miscarriage, and 61 cases (61%), resulted in failed IVF cycle. SOD, LPO, CAT, and TAP levels in the endometrial secretions of the three groups were statistically different (P-value <0.01, <0.001, <0.001, and <0.001, respectively). TTG levels in endometrial secretion of three groups were not statistically different (P-value=0.837). Our results indicated that higher levels of antioxidants such as SOD, CAT, or TAP, and lower levels of oxidative stress markers such as LPO in the endometrial secretions were associated with successful IVF outcome. PMID:27232354

  12. Effects of high-dose and multiple-dose gonadotropin stimulation on mouse oocyte quality as assessed by preimplantation development following in vitro fertilization.

    PubMed

    Edgar, D H; Whalley, K M; Mills, J A

    1987-10-01

    The effects of increasing the level of ovarian stimulation on preimplantation embryonic development were assessed using a mouse in vitro fertilization system. When F1 hybrid (C57BL/6 X CBA/Ca) mice received a single injection of 5 IU pregnant mare's serum gonadotropin (PMSG) followed 60 hr later by 5 IU human chorionic gonadotropin (hCG) approximately 50% of the resultant postovulatory oocytes developed to the blastocyst stage following in vitro fertilization. Increasing the single dose of PMSG to 10 or 15 IU resulted in significant reductions in the frequency of development to the blastocyst stage. When one or two additional doses of 5 IU PMSG were administered 24 and 48 hr after an initial injection of 5 IU, lower frequencies of oocytes with the potential for full preimplantation development were again observed. This reduction in gamete quality was significantly greater when the final dose of PMSG was administered only 12 hr prior to hCG. The results suggest that excessive gonadotropin stimulation may compromise the quality of the preimplantation embryos obtained following in vitro fertilization and that the timing of gonadotropin administration may also be critical.

  13. On teaching biopolicy and values in selected reproductive technologies: abortion, in vitro fertilization, and surrogate motherhood.

    PubMed

    Moreland, L B

    1986-08-01

    This article is divided into three parts: "Faculty Preparation: Immersion in the Literature," "Conceptualizing and Organizing the Course," and "Course Design." The first part is addressed particularly to readers without access to a computer search of the literature. It suggests resource materials for each of the technologies. It also speaks to the meanings of and approaches which may be used to study biopolitics. Biopolicy is the approach used in the course. The second part addresses the problem of finding a unifying concept which would bring cohesion to the multiple and diverse materials and issues. It also includes the statement of purpose and course objectives. It is here that the development of identifying and demonstrating the nexus between the reproductive technologies and public policy begins. It also discusses methodologies used in the course. The third part lists the activities that served as a guide in designing the course and cites remaining caveats in course development. The paper concludes with statements about required reading materials, review materials, audio-visual aids, and a broad outline of the course. PMID:17153268

  14. Protective Effect of Royal Jelly on In Vitro Fertilization (IVF) in Male Mice Treated with Oxymetholone

    PubMed Central

    Zahmatkesh, Ensieh; Najafi, Gholamreza; Nejati, Vahid

    2015-01-01

    Objective This study aimed to investigate the effects of royal jelly (RJ) on catalase, total antioxidant capacity and embryo development in adult mice treated with oxymetholone (OXM). Materials and Methods In this exprimental study, 32 male and 96 female adult Naval Medical Research Institute (NMRI) mice (7-9 weeks of age) with a ratio of 1:3 for fertili- zation purposes were randomly divided into 4 groups as follows: i. Control group (n=8) receiving 0.1 ml/mice saline daily by gavage for 30 day, ii. RJ group (n=8) treated with RJ at a dose of 100 mg/kg daily by gavage for 30 days, iii. OXM group (n=8) receiving OXM at the dose of 5 mg/kg daily by gavage for 30 days and iv. RJ+OXM group (n=8) receiving RJ at the dose of 100 mg/kg daily by gavage concomitant with 100 mg/kg OXM adminis- tration for 30 days. Results Analysis revealed a significant reduction in catalase, total antioxidant, as well as embryo development in OXM group (P<0.05). However, RJ group showed a salient recovery in the all of the above mentioned parameters and embryo toxicity. Conclusion The results of this study indicated a partially protective effect of RJ against OXM-induced embryo toxicity. PMID:26464831

  15. The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI.

    PubMed

    Johnson, L A; Welch, G R; Rens, W

    1999-01-01

    The Beltsville sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow-cytometric separation of X- and Y-chromosome-bearing sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow-cytometrically sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep-uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted sperm populations and by fluorescence in situ hybridization and PCR of individual sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for sperm sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y sperm from about .35 million/h to 5 or 6 million sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted sperm. PMID:15526798

  16. Laminin-111 Inhibits Bovine Fertilization but Improves Embryonic Development in vitro, and Receptor Integrin-β1 is Involved in Sperm-Oocyte Binding.

    PubMed

    Lin, F; Huang, C-J; Liu, C-S; Guo, L-L; Liu, G; Liu, H-J

    2016-10-01

    This study detected the distribution of laminin during embryonic formation by immunofluorescence. To determine the possible function of laminin on developmental ability of in vitro fertilized embryos, the presumptive zygotes were divided and transferred to CR1aa medium supplemented with different concentrations (0 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml) of laminin. To explore the association with sperm-oocyte fusion, oocytes and/or sperm were pre-incubated with laminin or anti-β1 antibody before insemination. Laminin was absent in mature oocytes and could be detected first at the 8-cell stage and then displayed an increasing tendency. Adding 10 μg/ml laminin to the culture medium improved embryonic development including cleavage rate, blastocyst rate, total cell numbers in the blastocyst and cell numbers in the inner cell mass. Laminin inhibited sperm-oocyte fusion when incubated with oocytes and/or sperm before in vitro fertilization, and only integrin-β1 of sperm was involved in sperm-oocyte binding. Inhibition may be caused by blocking β1, but why laminin inhibits fertilization is still unknown. The results suggest that laminin plays an important role during embryonic formation and has a negative function in sperm-oocyte fusion, but improves embryonic development. However, only integrin-β1 is involved in sperm-oocyte binding. PMID:27491353

  17. Ultrastructural comparison of porcine putative embryonic stem cells derived by in vitro fertilization and somatic cell nuclear transfer

    PubMed Central

    YOO, Hyunju; KIM, Eunhye; HWANG, Seon-Ung; YOON, Junchul David; JEON, Yubyeol; PARK, Kyu-Mi; KIM, Kyu-Jun; JIN, Minghui; LEE, Chang-Kyu; LEE, Eunsong; KIM, Hyunggee; KIM, Gonhyung; HYUN, Sang-Hwan

    2016-01-01

    The ultrastructure of porcine putative embryonic stem cells and porcine fetal fibroblasts (PFFs) was analyzed by transmission electron microscopy. The aim of this study was to compare the features of organelles in in vitro fertilization (IVF) derived porcine embryonic stem cells (IVF-pESCs) and somatic cell nuclear transfer (SCNT) derived pESCs (SCNT-pESCs). Also, the features of organelles in high-passage IVF-pESCs were compared with those in low-passage cells. The ultrastructure of PFFs showed rare microvilli on the cell surfaces, polygonal or irregular nuclei with one to two reticular-shaped nucleoli and euchromatin, low cytoplasm-to-nucleus ratios, rare ribosomes, rare rough endoplasmic reticulum, elongated mitochondria, rich lysosomes and rich phagocytic vacuoles. IVF-pESCs showed rare microvilli on the cell surfaces, round or irregular nuclei with one to two reticular-shaped nucleoli and euchromatin, low cytoplasm-to-nucleus ratios, rich ribosomes, long stacks of rough endoplasmic reticulum, elongated mitochondria, rare lysosomes and rare autophagic vacuoles. By contrast, SCNT-pESCs showed rich microvilli with various lengths and frequencies on the cell surfaces, polygonal nuclei with one reticular shaped nucleoli and heterochromatin, high cytoplasm-to-nucleus ratios, rare ribosomes, rare rough endoplasmic reticulum, round mitochondria, rich lysosomes and rich phagocytic vacuoles with clear intercellular junctions. Furthermore, high-passage IVF-pESCs showed irregularly shaped colonies, pyknosis and numerous lysosomes associated with autophagic vacuoles showing signs of apoptosis. In conclusion, this study confirms that the ultrastructural characteristics of pESCs differ depending on their origin. These ultrastructural characteristics might be useful in biomedical research using pESCs, leading to new insights regarding regenerative medicine and tissue repair. PMID:26821870

  18. The use of synthetic culture medium and patient serum for human in vitro fertilization and embryo replacement.

    PubMed

    Feichtinger, W; Kemeter, P; Menezo, Y

    1986-04-01

    The use of heat-inactivated patient serum as both fertilization and embryo replacement medium was compared in a prospective randomized study with a fully synthetic culture medium containing human serum albumin without serum addition (B3 INRA Menezo). Another series of the author's IVF program was analyzed retrospectively when a commercially available synthetic medium with bovine serum albumin (B2 INRA Menezo) or B3, as mentioned above, was used without serum addition for fertilization but with 50-100% patient serum as embryo replacement medium. Fertilization rates were significantly higher in the synthetic culture media (70%) than in serum (57%). The rate of polyploid fertilization was significantly lowest in B3 medium. There was a clear trend toward better pregnancy rates when high-percentage or 100% patient serum was used for embryo replacement, no matter if one, two, three, four or more embryos were replaced. We conclude that there should be no need for any kind of serum addition to fertilization media. The present study proves our previous observation that the use of serum seems to be beneficial as embryo replacement medium. This might well be explained by a "protein stick effect" due to the high macromolecular contents of serum rather than by viscosity measurements, since no significant increase in viscosity was observed when a high percentage of serum was added to culture media.

  19. Cell-free DNA in Human Follicular Microenvironment: New Prognostic Biomarker to Predict in vitro Fertilization Outcomes.

    PubMed

    Traver, Sabine; Scalici, Elodie; Mullet, Tiffany; Molinari, Nicolas; Vincens, Claire; Anahory, Tal; Hamamah, Samir

    2015-01-01

    Cell-free DNA (cfDNA) fragments, detected in blood and in other biological fluids, are released from apoptotic and/or necrotic cells. CfDNA is currently used as biomarker for the detection of many diseases such as some cancers and gynecological and obstetrics disorders. In this study, we investigated if cfDNA levels in follicular fluid (FF) samples from in vitro fertilization (IVF) patients, could be related to their ovarian reserve status, controlled ovarian stimulation (COS) protocols and IVF outcomes. Therefore, 117 FF samples were collected from women (n = 117) undergoing IVF/Intra-cytoplasmic sperm injection (ICSI) procedure and cfDNA concentration was quantified by ALU-quantitative PCR. We found that cfDNA level was significantly higher in FF samples from patients with ovarian reserve disorders (low functional ovarian reserve or polycystic ovary syndrome) than from patients with normal ovarian reserve (2.7 ± 2.7 ng/μl versus 1.7 ± 2.3 ng/μl, respectively, p = 0.03). Likewise, FF cfDNA levels were significant more elevated in women who received long ovarian stimulation (> 10 days) or high total dose of gonadotropins (≥ 3000 IU/l) than in women who received short stimulation duration (7-10 days) or total dose of gonadotropins < 3000 IU/l (2.4 ± 2.8 ng/μl versus 1.5 ± 1.9 ng/μl, p = 0.008; 2.2 ± 2.3 ng/μl versus 1.5 ± 2.1 ng/μl, p = 0.01, respectively). Finally, FF cfDNA level was an independent and significant predictive factor for pregnancy outcome (adjusted odds ratio = 0.69 [0.5; 0.96], p = 0.03). In multivariate analysis, the Receiving Operator Curve (ROC) analysis showed that the performance of FF cfDNA in predicting clinical pregnancy reached 0.73 [0.66-0.87] with 88% specificity and 60% sensitivity. CfDNA might constitute a promising biomarker of follicular micro-environment quality which could be used to predict IVF prognosis and to enhance female infertility management. PMID:26288130

  20. Assisted ejaculation and in-vitro fertilization in the treatment of infertile spinal cord-injured men: the role of intracytoplasmic sperm injection.

    PubMed

    Hultling, C; Rosenlund, B; Levi, R; Fridström, M; Sjöblom, P; Hillensjö, T

    1997-03-01

    The objective of the present longitudinal descriptive study was to extend previous observations on the benefit of in-vitro fertilization (IVF) in cases of anejaculatory infertility due to spinal cord injuries (SCI) and to report results achieved by intracytoplasmic sperm injection (ICSI). The study was performed in a national referral unit for SCI, Spinalis SCI Research Unit, the Karolinska Institute. The patient material consisted of couples with SCI men seeking treatment for their infertility. The inclusion criteria were: stable relationship, motile spermatozoa in a diagnostic sample and no female contraindications. Spermatozoa were retrieved through electroejaculation or vibratory stimulation. If the sperm quality was judged to be sufficient, standard IVF was performed. ICSI was employed if the semen quality was extremely poor. We have treated 25 couples in 52 cycles, leading to 81 ovum retrievals and 47 embryo transfers. Total sperm counts were very variable (0.01-978 x 10(6)). Before the introduction of ICSI the fertilization rate was 30%. ICSI increased the fertilization rate to 88%. There was no association between the pregnancy rate and the sperm count, level of injury or fertilization technique. A total of 16 clinical pregnancies was established, leading to 11 deliveries. This gives a cumulative pregnancy rate per couple of 56%.

  1. Effect of recombinant human follicle-stimulating hormone and luteinizing hormone on in vitro maturation of porcine oocytes evaluated by the subsequent in vitro development of embryos obtained by in vitro fertilization, intracytoplasmic sperm injection, or parthenogenetic activation.

    PubMed

    Silvestre, M A; Alfonso, J; García-Mengual, E; Salvador, I; Duque, C C; Molina, I

    2007-05-01

    The aim of this work was to study the effect of recombinant human (rh) FSH and LH on in vitro maturation of pig oocytes compared with a conventional hormonal supplement based on equine (PMSG) and human chorionic gonadotropins (hCG), as evaluated by the developmental ability of 3 types of pig embryos obtained by in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), or artificial activation (ATA). In Exp. 1, one cumulus-oocyte complex group (A group) was supplemented with rh-FSH and rh-LH (0.1 IU/mL each), and the other group (B group) was supplemented with PMSG and hCG (10 IU/mL each). No differences in nuclear maturation between the A and B groups were observed (68.5 vs. 71.4%, respectively). No differences were detected between hormonal treatments in the rates of cleavage or blastocyst formation of ATA, IVF, and ICSI embryos. Total cell number of the embryos was not significantly different in any experimental group (A: 31.1, 28.5, and 19.8 vs. B: 25.2, 25.5, and 20.6 for ATA, IVF, and ICSI embryos, respectively). In Exp. 2, the effects of different concentrations of rh-FSH and rh-LH (0.5, 0.1, or 0.05 IU/mL) in maturation medium on nuclear maturation and in vitro development of embryos obtained by IVF were studied. No effect of different hormonal concentrations on blastocyst formation rates was observed (8.5, 13.0, and 5.7%, respectively). Blastocyst cell number was not different in any experimental group. In conclusion, the results obtained here permit us to substitute PMSG and hCG with rh-FSH and rh-LH and to produce pig embryos obtained by IVF, ICSI, or ATA.

  2. Transabdominal follicular aspiration in an in vitro fertilization cycle: experiences with an unusual but necessary intervention in a resource-limited setting

    PubMed Central

    Osemwenkha, Abieyuwa

    2016-01-01

    Controlled ovarian hyperstimulation is one of the major steps of in vitro fertilization. The inaccessibility or non-visualization of developing follicles on transvaginal sonography (the preferred imaging method) may be misjudged as a poor response, resulting in cycle cancellation. It is necessary to scrupulously appraise proxy indicators for ovarian response, such as estradiol levels, endometrial thickness, and other individual clinical characteristics. This can prompt meticulous transabdominal ultrasound follicular monitoring and oocyte retrieval with the goal of averting cycle cancellation and improving treatment outcomes. PMID:27104159

  3. Egg recovery completed with a “manually-created” negative pressure is still an option in cases of emergency or “low-cost” in vitro fertilization?

    PubMed Central

    Kalampokas, Theodoros; Maheswari, Abha

    2015-01-01

    This is a report of a case of an egg recovery procedure completed with manual suction instead of the automated negative pressure suction. A 35–year-old woman who was undergoing in vitro fertilization (IVF) treatment had to undergo oocyte recovery using manual suction through a syringe instead of the automated negative pressure suction systems because of the failure of both the initial and replacement systems. The treatment cycle ended in a positive pregnancy test and a clinical pregnancy with acceptable oocyte fertilization rates and no complications during procedure. The case presented may be an implication for an alternative implementation in cases of emergency, particularly when low-cost IVF is a target. PMID:26692778

  4. TP53 PIN3 and PEX4 polymorphisms and infertility associated with endometriosis or with post-in vitro fertilization implantation failure

    PubMed Central

    Paskulin, D D; Cunha-Filho, J S L; Souza, C A B; Bortolini, M C; Hainaut, P; Ashton-Prolla, P

    2012-01-01

    p53 has a crucial role in human fertility by regulating the expression of leukemia inhibitory factor (LIF), a secreted cytokine critical for blastocyst implantation. To examine whether TP53 polymorphisms may be involved with in vitro fertilization (IVF) failure and endometriosis (END), we have assessed the associations between TP53 polymorphism in intron 2 (PIN2; G/C, intron 2), PIN3 (one (N, non-duplicated) or two (D, duplicated) repeats of a 16-bp motif, intron 3) and polymorphism in exon 4 (PEX4; C/G, p.P72R, exon 4) in 98 women with END and 115 women with post-IVF failure. In addition, 134 fertile women and 300 women unselected with respect to fertility-related features were assessed. TP53 polymorphisms and haplotypes were identified by amplification refractory mutation system polymerase chain reaction. TP53 PIN3 and PEX4 were associated with both END (P=0.042 and P=0.007, respectively) and IVF (P=0.004 and P=0.009, respectively) when compared with women both selected and unselected for fertility-related features. Haplotypes D-C and N-C were related to higher risk for END (P=0.002, P=0.001, respectively) and failure of IVF (P=0.018 and P=0.002, respectively) when compared with the Fertile group. These results support that specific TP53 haplotypes are associated with an increased risk of END-associated infertility and with post-IVF failure. PMID:23013791

  5. TP53 PIN3 and PEX4 polymorphisms and infertility associated with endometriosis or with post-in vitro fertilization implantation failure.

    PubMed

    Paskulin, D D; Cunha-Filho, J S L; Souza, C A B; Bortolini, M C; Hainaut, P; Ashton-Prolla, P

    2012-01-01

    p53 has a crucial role in human fertility by regulating the expression of leukemia inhibitory factor (LIF), a secreted cytokine critical for blastocyst implantation. To examine whether TP53 polymorphisms may be involved with in vitro fertilization (IVF) failure and endometriosis (END), we have assessed the associations between TP53 polymorphism in intron 2 (PIN2; G/C, intron 2), PIN3 (one (N, non-duplicated) or two (D, duplicated) repeats of a 16-bp motif, intron 3) and polymorphism in exon 4 (PEX4; C/G, p.P72R, exon 4) in 98 women with END and 115 women with post-IVF failure. In addition, 134 fertile women and 300 women unselected with respect to fertility-related features were assessed. TP53 polymorphisms and haplotypes were identified by amplification refractory mutation system polymerase chain reaction. TP53 PIN3 and PEX4 were associated with both END (P=0.042 and P=0.007, respectively) and IVF (P=0.004 and P=0.009, respectively) when compared with women both selected and unselected for fertility-related features. Haplotypes D-C and N-C were related to higher risk for END (P=0.002, P=0.001, respectively) and failure of IVF (P=0.018 and P=0.002, respectively) when compared with the Fertile group. These results support that specific TP53 haplotypes are associated with an increased risk of END-associated infertility and with post-IVF failure. PMID:23013791

  6. Birth of a western lowland gorilla (Gorilla gorilla gorilla) following in vitro fertilization and embryo transfer.

    PubMed

    Pope, C E; Dresser, B L; Chin, N W; Liu, J H; Loskutoff, N M; Behnke, E J; Brown, C; McRae, M A; Sinoway, C E; Campbell, M K; Cameron, K N; Owens, O M; Johnson, C A; Evans, R R; Cedars, M I

    1997-01-01

    A 21-year-old multiparous female exhibiting 31-41 day menstrual cycles was given hFSH (225 IU/day, Metrodin 75, from cycle day 3 through 9 (menses = day 1) and hCG (10,000 IU, Profasi, on day 10 to stimulate follicular development. At 35 h after hCG, under isoflurane (AErrane) anesthesia, follicles were aspirated by controlled suction under transvaginal ultrasound guidance. Metaphase II oocytes (n = 11) were placed in modified human tubal fluid (mHTF, 100 microliters) medium under oil at 37 degrees C in humidified 5% CO2. Frozen semen, collected by voluntary ejaculation, was thawed (70 degrees C H2O bath, 6 sec), diluted slowly, centrifuged, and resuspended in mHTF, and 160,000 motile spermatozoa/ml were added at 6 h after oocyte recovery. At 21 h postinsemination (p.i.) eight oocytes were at the two-cell stage, five were cryopreserved, and three were cultured to the six- to eight-cell stage in mHTF with granulosa cells before transcervical uterine transfer at 47 h p.i. using a Teflon catheter. Micronized progesterone (400 mg/d) was orally administered for 10 weeks posttransfer (p.t.). Ultrasound examination revealed a single fetus at 15 weeks p.t., and unassisted delivery of a live 1.37 kg female infant occurred at 29 weeks. Am. J. Primatol. 41:247-260, 1997.

  7. Cryopreservation and in vitro fertilization at the Zebrafish International Resource Center

    PubMed Central

    Carmichael, Carrie; Westerfield, Monte; Varga, Zoltán M.

    2009-01-01

    In recent decades, laboratories throughout the world generated several thousand mutant, transgenic, and wild-type zebrafish lines and more lines continue to be produced. At the same time, relatively little effort has been expended to develop reliable, high-throughput, standardized, long-term cryopreservation storage methods, even though laboratories and the research community as a whole struggle to maintain the large number of lines alive. Safe and reliable methods for maintaining these valuable genetic resources are vital for future biomedical research. Cryopreservation is the most efficient method for large-scale, long-term storage of important genetic materials. It extends the time offspring can be produced from individual fish, reduces the need to maintain live populations, and can prevent catastrophic loss of irreplaceable research lines. Cryopreservation is also the most cost-effective alternative for maintaining genetic resources because it reduces costs for animal and facility maintenance, personnel, and space. In addition, it provides novel opportunities to develop new types of research using large numbers of lines. For example, several genetic strategies, such as TILLING or enhancer and gene trapping, depend on the use of cryopreservation to bypass generations of live organisms until a strain is revived for research. In this chapter, we describe and discuss the current cryopreservation method used at the Zebrafish International Resource Center. This method is derived from the initial protocol developed for zebrafish over 20 years ago that has recently been refined (1). PMID:19378097

  8. Impact of Whole Systems Traditional Chinese Medicine on In Vitro Fertilization Outcomes

    PubMed Central

    Hullender Rubin, Lee E.; Opsahl, Michael S.; Wiemer, Klaus; Mist, Scott D.; Caughey, Aaron B.

    2015-01-01

    Patients undergoing IVF patients may receive either acupuncture or whole-systems traditional Chinese medicine (WS-TCM) as an adjuvant IVF treatment. WS-TCM is a complex intervention that can include acupuncture, Chinese herbal medicine, dietary, lifestyle recommendations, or both. In this retrospective cohort study, 1231 IVF patient records were reviewed to assess the effect of adjuvant WS-TCM on IVF outcomes compared among three groups: IVF with no additional treatment; IVF and elective acupuncture on day of embryo transfer; or IVF and elective WS-TCM. The primary outcome was live birth. Of 1069 non-donor cycles, WS-TCM was associated with greater odds of live birth compared with IVF alone (adjusted odds ratio [AOR] 2.09; 95% confidence interval [CI] 1.36 to 3.21), or embryo transfer with acupuncture only (AOR 1.62; 95% CI 1.04 to 2.52). Of 162 donor cycles, WS-TCM was associated with increased live births compared with all groups (odds Ratio [OR] 3.72; 95% CI 1.05 to 13.24, unadjusted] or embryo transfer with acupuncture only (OR 4.09; 95% CI: 1.02 to 16.38, unadjusted). Overall, IVF with adjuvant WS-TCM was associated with greater odds of live birth in donor and non-donor cycles. These results should be taken cautiously as more rigorous research is needed. PMID:25911598

  9. Milder is better? advantages and disadvantages of "mild" ovarian stimulation for human in vitro fertilization

    PubMed Central

    2011-01-01

    In the last decades, several steps have been made aiming at rendering human IVF more successful on one side, more tolerable on the other side. The "mild" ovarian stimulation approach, in which a lower-than-average dose of exogenous gonadotropins is given and gonadotropin treatment is started from day 2 to 7 of the cycle, represents a significant step toward a more patient's friendly IVF. However, a clear view of its virtues and defects is still lacking, because only a few prospective randomized trials comparing "mild" vs. conventional stimulation exist, and they do not consider some important aspects, such as, e.g., thawing cycles. This review gives a complete panorama of the "mild" stimulation philosophy, showing its advantages vs. conventional ovarian stimulation, but also discussing its disadvantages. Both patients with a normal ovarian responsiveness to exogenous gonadotropins and women with a poor ovarian reserve are considered. Overall, we conclude that the level of evidence supporting the use of "mild" stimulation protocols is still rather poor, and further, properly powered prospective studies about "mild" treatment regimens are required. PMID:21324155

  10. Impact of whole systems traditional Chinese medicine on in-vitro fertilization outcomes.

    PubMed

    Hullender Rubin, Lee E; Opsahl, Michael S; Wiemer, Klaus E; Mist, Scott D; Caughey, Aaron B

    2015-06-01

    Patients undergoing IVF may receive either acupuncture or whole-systems traditional Chinese medicine (WS-TCM) as an adjuvant IVF treatment. WS-TCM is a complex intervention that can include acupuncture, Chinese herbal medicine, dietary, lifestyle recommendations. In this retrospective cohort study, 1231 IVF patient records were reviewed to assess the effect of adjuvant WS-TCM on IVF outcomes compared among three groups: IVF with no additional treatment; IVF and elective acupuncture on day of embryo transfer; or IVF and elective WS-TCM. The primary outcome was live birth. Of 1069 non-donor cycles, WS-TCM was associated with greater odds of live birth compared with IVF alone (adjusted odds ratio [AOR] 2.09; 95% confidence interval [CI] 1.36 to 3.21), or embryo transfer with acupuncture only (AOR 1.62; 95% CI 1.04 to 2.52). Of 162 donor cycles, WS-TCM was associated with increased live births compared with all groups (odds Ratio [OR] 3.72; 95% CI 1.05 to 13.24, unadjusted) or embryo transfer with acupuncture only (OR 4.09; 95% CI: 1.02 to 16.38, unadjusted). Overall, IVF with adjuvant WS-TCM was associated with greater odds of live birth in donor and non-donor cycles. These results should be taken cautiously as more rigorous research is needed.

  11. Successful twin pregnancy in a dual-transplant couple resulting from in-vitro fertilization and intracytoplasmic sperm injection: case report.

    PubMed

    Case, A M; Weissman, A; Sermer, M; Greenblatt, E M

    2000-03-01

    There are numerous reports of successful pregnancy following liver transplantation. Little information is available regarding the incidence and management of infertility in transplant recipients, particularly the use of artificial reproductive technologies. We present a case of a successful twin pregnancy resulting from in-vitro fertilization with intracytoplasmic sperm injection (IVF/ICSI) in a liver transplant recipient, whose partner was a renal transplant recipient with severe oligozoospermia. With careful evaluation and monitoring, and the involvement of appropriate consultants, artificial reproductive technologies can be safely used in transplant recipient couples experiencing infertility.

  12. An 18-month survey of infertility treatment by in vitro fertilization, gamete and zygote intrafallopian transfer, and replacement of frozen-thawed embryos.

    PubMed

    Staessen, C; Camus, M; Khan, I; Smitz, J; Van Waesberghe, L; Wisanto, A; Devroey, P; Van Steirteghem, A C

    1989-02-01

    An 18-month survey of infertility treatment by in vitro fertilization (IVF) and related procedures at the Centre for Reproductive Medicine of the Vrije Universiteit Brussel is described. During this period, 1326 treatment cycles were started in patients with long-standing infertility and 1135 oocyte retrievals were performed in 771 different patients. IVF and embryo transfer (ET) after laparoscopic (N = 793) or ultrasonically guided (N = 342) ovum pickup, gamete intrafallopian transfer (GIFT; N = 284), or zygote intrafallopian transfer (ZIFT; N = 15) combined with IVF as well as the replacement of cryopreserved embryos yielded an overall pregnancy rate of 21.8% per started cycle. Echographic and laparoscopic oocyte retrieval gave similar results except for a higher fertilization rate after echographic-guided retrieval. For in vitro fertilization and embryo transfer an overall pregnancy rate of 26% per transfer was obtained. For GIFT and ZIFT the pregnancy rates were, respectively, 27.8 and 46.7% per replacement. For each procedure one-third of the pregnancies aborted. After the replacement of frozen and thawed embryos, during a natural cycle, a significantly lower fetal loss was observed.

  13. Global gene transcription patterns in in vitro-cultured fertilized embryos and diploid and haploid murine parthenotes

    SciTech Connect

    Cui Xiangshun; Li Xingyu; Kim, Nam-Hyung . E-mail: nhkim@chungbuk.ac.kr

    2007-01-19

    To gain insights into the roles the paternal genome and chromosome number play in pre-implantation development, we cultured fertilized embryos and diploid and haploid parthenotes (DPs and HPs, respectively), and compared their development and gene expression patterns. The DPs and fertilized embryos did not differ in developmental ability but HPs development was slower and characterized by impaired compaction and blastocoel formation. Microarray analysis revealed that fertilized blastocysts expressed several genes at higher levels than DP blastocysts; these included the Y-chromosome-specific gene eukaryotic translation initiation factor 2, subunit 3, structural gene Y-linked (Eif2s3y) and the imprinting gene U2 small nuclear ribonucleoprotein auxiliary factor 1, related sequence 1 (U2af1-rs1). We also found that when DPs and HPs were both harvested at 44 and 58 h of culture, they differed in the expression of 38 and 665 genes, respectively. However, when DPs and HPs were harvested at the midpoints of 4-cell stage (44 and 49 h, respectively), no differences in expression was observed. Similarly, when the DPs and HPs were harvested when they became blastocysts (102 and 138 h, respectively), only 15 genes showed disparate expression. These results suggest that while transcripts needed for early development are delayed in HPs, it does progress sufficiently for the generation of the various developmental stages despite the lack of genetic components.

  14. Fertility differentials in Nigeria.

    PubMed

    Ogum, G E

    1980-01-01

    A review of the numerous fertility studies carried out in Nigeria during the last 3 decades. Evidence of differences in fertility among subgroups of the population is gleaned from these studies, as very few have addressed this issue specifically. Results still need to be evaluated. The only strong indication of possible geographic fertility differentials remains Van de Walle's analysis of age data from the census of 1952-53. 2 studies among populations of southwestern Nigeria yielded conflicting results with regard to urban-rural differentials, 1 finding lower fertility among urban residents while the other found no differences. Similarly, 2 studies with regard to religious differences yielded somewhat conflicting results, 1 showing Moslems and traditional worshippers having higher fertility than Christians, the other showing Moslems with lower fertility than traditional believers. There is some consensus among researchers that education has some depressing effect on fertility.

  15. Separation of motile sperm for in vitro fertilization from frozen-thawed bull semen using progesterone induction on a microchip.

    PubMed

    Li, Jingchun; Ning, Bolin; Cao, Xinyan; Luo, Yinghua; Guo, Li; Wei, Guosheng; Liu, Shengjun; Zhang, Ying; Zhang, Aizhong; Wu, Rui; Li, Yanbing

    2016-09-01

    This study presents a novel method for the separation of motile sperm from non-progressive motile and immotile sperm and in vitro Fertilization (IVF). This separation of bull sperm was accomplished by inducing chemotaxis along a progesterone release agent in a 7.5-mm microchannel microchip composed of a biocompatible polydimethysiloxane layer and a glass gradient. The selected sperm was applied directly for IVF. In the first experiment, we tested the effect of different lengths of microchannnel (5mm, 7.5mm and 10mm) on quality parameter of separated sperm. The results showed that separated sperm using 7.5-mm microchannel chip were improved in sperm motility, swimming velocity, and beat frequency compared with other groups. In the second experiment, a medium containing sperm from swim-up method and outlet reservoir of our 7.5-mm microchannel chip was collected and mitochondrial activity of the sperm was determined by fluorescence microscopy. The sperm from the microchip had higher mitochondria activity (47.6%±6.0%) than the sperm from the swim-up method (23.6%±4.7%) (P<0.05). There were significant differences in rate of acrosome intactness between the swim-up method and the microchip (36.0%±4.1% vs. 66.8±2.1%, respectively, P<0.05). In the third experiment, we compared sperm penetration in the microchip-IVF system with a standard IVF method (droplet-IVF). The microchip-IVF group had the highest percentages of oocytes penetrated (82.2%±1.6% vs. 63.5%±2.4%) and monospermic oocytes (67.8%±3.4% vs. 42.4%±1.5%). In addition, early developmental competence of oocytes to the blastocyst stage was higher when the oocytes were inseminated in the microchip-IVF system compared with those inseminated in a standard droplet-IVF system. These results demonstrate that our microchip based on a sperm chemotaxis system is useful for motile sperm separation from frozen-thawed bull semen for IVF. Therefore, the optimized microchip system provides a good opportunity to sort

  16. Efficacy of growth hormone supplementation with gonadotrophins in vitro fertilization for poor ovarian responders: an updated meta-analysis.

    PubMed

    Yu, Xiaoying; Ruan, Jian; He, Lian-Ping; Hu, Weihua; Xu, Qinyang; Tang, Jingwen; Jiang, Jian; Han, Jun; Peng, Yi-Feng

    2015-01-01

    Growth hormone (GH) is involved in the regulation of male and female infertility. Several clinical studies reveal that adjuvant GH treatment has a possible role in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), especially in poor ovarian responders (POR) undergoing IVF/ICSI. Recent studies suggest that GH addition in POR patients significantly improves the rate of clinical pregnancy and live birth. Databases including PubMed, Embase, the Cochrane Central China National Knowledge Infrastructure (CNKI) and Google Scholar were searched for randomized controlled trials (RCTs) or controlled clinical trials (CCTs) on the effectiveness of GH supplementation with gonadotrophins in IVF/ICSI for POR. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data, and assessed methodological quality. Meta Analyst Beta 3.13 software was used to meta-analysis. Eleven studies (six RCTs and five CCTs) and 3788 subjects (613 subjects in cases group and 3175 subjects in controls group) were included in our study. The results of meta-analysis showed that GH addition significantly increased serum E2 level on the day of HCG (OR = 0.55; 95% CI = 0.127-0.973) and MII oocyte number (OR = 0.827; 95% CI = 0.470-1.184). Furthermore, GH addition significantly improved the number of 2PN (OR = 0.934; 95% CI = 0.206-1.661) and obtained embryos (OR = 0.934; 95% CI = 0.206-1.661). However, no significant difference was found for the overall implantation rate was 8.8% (95% CI = -0.062-0.237) and clinical pregnancy rate was 5.1% (95% CI = -0.033-0.134). The present result revel that GH supplementation for IVF/ICSI in POR increases the probability of serum E2 level on the day of HCG, the number of MII oocyte, 2PN and obtained embryos. However, GH addition does not increase implantation rate and clinical pregnancy rates. Due to the limited quantity and quality of the included studies as well as the difference in methodology, we

  17. Birth after human chorionic gonadotropin-primed oocyte in vitro maturation and fertilization with testicular sperm in a normo-ovulatory patient

    PubMed Central

    González-Ortega, Claudia; Piña-Aguilar, Raul Eduardo; Cancino-Villareal, Patricia; Gutiérrez-Gutiérrez, Antonio Martin

    2016-01-01

    In this report, we present a case of in vitro maturation (IVM) with surgical retrieved testicular sperm in a normo-ovulatory female. Human chorionic gonadotropin-primed IVM, testicular biopsy for sperm retrieval and intracytoplasmic sperm injection with fresh sperm were performed. Fourteen cumulus-oocyte complexes were obtained in germinal vesicle or metaphase I stage, eight oocytes reached metaphase II, seven presumptive zygotes were obtained, and three cleavage stages embryos in day 2 were transferred producing a singleton pregnancy. A single healthy newborn was obtained. Our results suggest that IVM may be an alternative for in vitro fertilization in normo-ovulatory women even if surgical retrieval of sperm is needed. Further research is required to depict contributing factors to the success of IVM in indications different from polycystic ovaries syndrome and the role of male gamete. PMID:27803591

  18. Cycle scheduling for in vitro fertilization with oral contraceptive pills versus oral estradiol valerate: a randomized, controlled trial

    PubMed Central

    2013-01-01

    Background Both oral contraceptive pills (OCPs) and estradiol (E2) valerate have been used to schedule gonadotropin-releasing hormone (GnRH) antagonist in vitro fertilization (IVF) cycles and, consequently, laboratory activities. However, there are no studies comparing treatment outcomes directly between these two pretreatment methods. This randomized controlled trial was aimed at finding differences in ongoing pregnancy rates between GnRH antagonist IVF cycles scheduled with OCPs or E2 valerate. Methods Between January and May 2012, one hundred consecutive patients (nonobese, regularly cycling women 18–38 years with normal day 3 hormone levels and <3 previous IVF/ICSI attempts) undergoing IVF with the GnRH antagonist protocol were randomized to either the OCP or E2 pretreatment arms, with no restrictions such as blocking or stratification. Authors involved in data collection and analysis were blinded to group assignment. Fifty patients received OCP (30 μg ethinyl E2/150 μg levonorgestrel) for 12–16 days from day 1 or 2, and stimulation was started 5 days after stopping OCP. Similarly, 50 patients received 4 mg/day oral E2 valerate from day 20 for 5–12 days, until the day before starting stimulation. Results Pretreatment with OCP (mean±SD, 14.5±1.7 days) was significantly longer than with E2 (7.8±1.9 days). Stimulation and embryological characteristics were similar. Ongoing pregnancy rates (46.0% vs. 44.0%; risk difference, –2.0% [95% CI –21.2% to 17.3%]), as well as implantation (43.5% vs. 47.4%), clinical pregnancy (50.0% vs. 48.0%), clinical miscarriage (7.1% vs. 7.7%), and live birth (42.0% vs. 40.0%) rates were comparable between groups. Conclusions This is the first study to directly compare these two methods of cycle scheduling in GnRH antagonist cycles. Our results fail to show statistically significant differences in ongoing pregnancy rates between pretreatment with OCP and E2 for IVF with the GnRH antagonist protocol. Although the

  19. Preterm birth and low birth weight among in vitro fertilization singletons: a systematic review and meta-analyses.

    PubMed

    McDonald, Sarah D; Han, Zhen; Mulla, Sohail; Murphy, Kellie E; Beyene, Joseph; Ohlsson, Arne

    2009-10-01

    Our objective was to determine the risks of preterm birth (PTB) and low birth weight (LBW) in singletons conceived through in vitro fertilization (IVF)+/-intracytoplasmic sperm injection (ICSI) compared to spontaneously conceived singletons after matching or controlling for at least maternal age. The MOOSE guidelines for meta-analysis of observational studies were followed. Medline and Embase were searched using comprehensive search strategies. Bibliographies of identified articles were reviewed. English language studies examining LBW or PTB in singletons conceived by IVF or IVF/intracytoplasmic sperm injection, compared with spontaneously conceived singletons, that matched or controlled for at least maternal age. Two reviewers independently assessed titles, abstracts, full articles and study quality and extracted data. Dichotomous data were meta-analyzed using relative risks (RR) as measures of effect size with a random effects model and for continuous data weighted mean difference was calculated. Seventeen studies were included with 31,032 singletons conceived through IVF (+/-ICSI) and 81,119 spontaneously conceived singletons. After matching or controlling for maternal age and often other factors, compared to spontaneously conceived singletons, IVF singletons had increased risks of our two primary outcomes, PTB (RR 1.84, 95% CI 1.54, 2.21) and LBW (<2500 g, RR 1.60, 95% CI 1.29, 1.98). Singletons conceived through IVF or IVF/ICSI were at increased risk for late PTB (32-36 weeks, RR 1.52, 95% CI 1.01, 2.30), moderate PTB <32-33 weeks (RR 2.27, 95% CI 1.73, 2.97), very LBW (<1500 g, RR 2.65, 95% CI 1.83, 3.84), and intrauterine growth restriction (RR 1.45, 95% CI 1.04, 2.00), lower birth weights (-97 g, 95% CI -161 g, -33 g) and shorter mean gestations (-0.6 weeks, 95% CI -0.9 weeks, -0.4 weeks). In conclusion, IVF singletons have significantly increased risks of PTB, LBW and other adverse perinatal outcomes compared to spontaneously conceived singletons after

  20. Roles of the zona pellucida and functional exposure of the sperm-egg fusion factor 'IZUMO' during in vitro fertilization in pigs.

    PubMed

    Tanihara, Fuminori; Nakai, Michiko; Men, Nguyen Thi; Kato, Noriko; Kaneko, Hiroyuki; Noguchi, Junko; Otoi, Takeshige; Kikuchi, Kazuhiro

    2014-04-01

    The zona pellucida (ZP) is considered to play important roles in the prevention of polyspermy in mammalian oocytes. However, in pigs we have shown that the presence of the ZP accelerates sperm penetration into the ooplasm during in vitro fertilization (IVF). In the present study, we investigated the effects of the ZP on sperm binding, acrosomal status, and functional exposure of IZUMO, a critical factor involved in sperm-egg fusion, during IVF in pigs. We evaluated the numbers and acrosomal statuses of sperm binding to the ZP and oolemma, and being present in the ZP and perivitelline space (PVS) using ZP-intact and ZP-free oocytes. More sperm bound to the ZP than to the oolemma. The average number of sperm present in the PVS was 0.44-0.51 per oocyte, and all sperm had lost their acrosomes. The proportion of sperm that were immunopositive for anti-IZUMO antibody was significantly higher after they were passing or had passed through the ZP. Furthermore, addition of anti-IZUMO antibody to the fertilization medium significantly inhibited the penetration of sperm into ZP-free oocytes. These results suggest that, in pigs, the ZP induces the acrosome reaction, which is associated with the functional exposure of IZUMO, resulting in completion of fertilization.

  1. Estimation of the optimal timing of fertilization for embryo development of in vitro-matured bovine oocytes based on the times of nuclear maturation and sperm penetration.

    PubMed

    Koyama, Keisuke; Kang, Sung-Sik; Huang, Weiping; Yanagawa, Yojiro; Takahashi, Yoshiyuki; Nagano, Masashi

    2014-05-01

    The objective of this research was to estimate the optimal timing for fertilization to achieve proper embryonic development of in vitro-matured bovine oocytes. First, cumulus-oocyte complexes were subjected to in vitro maturation (IVM) for 14-22 hr. The timing when 50% of oocytes reached metaphase II stage was estimated to be 17.5 hr after IVM start. Next, using oocytes subjected to IVM for 12-30 hr, sperm penetration was examined after 4-18 hr of in vitro fertilization (IVF). A significant negative correlation between IVM duration and the timing when 50% of oocytes were penetrated by sperm after IVF start was observed (P<0.01). Finally, oocytes subjected to 12-30 hr of IVM were inseminated and cultured for 6 days to examine embryonic development. In the group with 22 hr of IVM, the percentages of cleaved embryos and blastocysts were the highest values in all groups. According to the regression equation describing the time from nuclear maturation to sperm penetration (x) and the percentage of blastocysts (y) (y=7.23x - 0.297x(2), P<0.01), the blastocyst rate peaked when sperm penetration occurred at 12.2 hr after achieving nuclear maturation. In conclusion, under the present IVM/IVF conditions, it was estimated that oocytes acquired their highest developmental competence at about 30 hr after IVM start, and thus, the optimal IVM duration was calculated to be about 21 hr.

  2. Absence of heat treatment of serum for culture medium supplementation does not adversely affect the outcome of in-vitro fertilization.

    PubMed

    Imoedemhe, D A; Sigue, A B; Pacpaco, E L; Olazo, A B; Luciano, E C

    1994-09-01

    This study was carried out to determine if not heat-treating serum prior to use for medium supplementation adversely affected in-vitro fertilization (IVF) of human oocytes. Morphologically mature human oocytes derived from 135 patients undergoing IVF treatment were studied. A total of 504 oocytes were incubated, inseminated and the resulting pronuclear oocytes cultured further in Earle's balanced salt solution (EBSS) supplemented with 10% non-heat-treated serum. Comparisons of fertilization rate and embryonic development were made between these and 687 control oocytes derived from the same patients but incubated, inseminated and resulting pronuclear oocytes cultured further in EBSS supplemented with 10% heat-treated serum. The fertilization rate of 74.4% (375/504) of oocytes handled in serum-supplemented medium that had not been heat-treated was significantly better than the rate of 67.7% (465/687) for controls (P < 0.0125). The proportion of pronucleate oocytes that cleaved was also significantly better in the non-heat-treated serum group: 270/300 (90%) versus 307/375 (81.8%) (P < 0.0025). There was no significant difference in the proportion of embryos with four or more cells at the time of embryo transfer. The results show that the absence of heat treatment of serum used to supplement culture medium has no adverse effect on the fertilization rate and short-term embryo development in vitro; hence we suggest that serum heat treatment is an unnecessary procedure and could be abandoned. PMID:7836531

  3. Sperm-egg interaction and functional assessment of springbok, impala and blesbok cauda epididymal spermatozoa using a domestic cattle in vitro fertilization system.

    PubMed

    Chatiza, F P; Bartels, P; Nedambale, T L; Wagenaar, G M

    2013-12-01

    The study assesses the possibility to estimate the potential fertility of post-thawed antelope (Antidorcas marsupialis), impala (Aepyceros melampus) and blesbok (Damaliscus dorcus phillipsi) epididymal sperm using homologous and heterologous IVF and the functioning of cattle IVF system to produce antelope embryos. Cauda epididymal sperm were collected from the antelope and cryopreserved under field conditions. In vitro matured domestic cow, blesbok and springbok oocytes were co-incubated in modified-Tyrode Lactate (m-TL) IVF media with springbok, impala and blesbok sperm for heterologous IVF and springbok and blesbok sperm for homologous IVF. A group of presumptive zygotes from each treatment were examined for sperm penetration and male pronuclear formation after 18h and the remainder were cultured and evaluated for embryo cleavage 22h later. The study shows that Modified Tyrode Lactate in vitro fertilization media supports survivability, capacitation and hyperactivation of springbok, impala and blesbok sperm. Springbok, impala and blesbok post-thawed epididymal spermatozoa are capable of fertilizing domestic cow oocytes under conditions that support domestic cattle IVF. Penetration, male pronuclear formation and embryo cleavage did not differ (p>0.05) between cow oocytes inseminated with sperm from springbok, impala or blesbok however these parameters were higher (p<0.05) for oocytes inseminated with bull sperm. Modified Tyrode Lactate IVF media supported homologous fertilization and embryo development in springbok and blesbok however did not support blastocyst development. These findings suggest that cattle provide a useful model for evaluating springbok, impala and blesbok post-thawed cauda epididymal sperm functionality. Domestic cattle embryo culture conditions need to be modified to promote blastosyst development in these antelope species. Such research provides an important tool in assisted reproductive technology development when high biological value

  4. Sperm-egg interaction and functional assessment of springbok, impala and blesbok cauda epididymal spermatozoa using a domestic cattle in vitro fertilization system.

    PubMed

    Chatiza, F P; Bartels, P; Nedambale, T L; Wagenaar, G M

    2013-12-01

    The study assesses the possibility to estimate the potential fertility of post-thawed antelope (Antidorcas marsupialis), impala (Aepyceros melampus) and blesbok (Damaliscus dorcus phillipsi) epididymal sperm using homologous and heterologous IVF and the functioning of cattle IVF system to produce antelope embryos. Cauda epididymal sperm were collected from the antelope and cryopreserved under field conditions. In vitro matured domestic cow, blesbok and springbok oocytes were co-incubated in modified-Tyrode Lactate (m-TL) IVF media with springbok, impala and blesbok sperm for heterologous IVF and springbok and blesbok sperm for homologous IVF. A group of presumptive zygotes from each treatment were examined for sperm penetration and male pronuclear formation after 18h and the remainder were cultured and evaluated for embryo cleavage 22h later. The study shows that Modified Tyrode Lactate in vitro fertilization media supports survivability, capacitation and hyperactivation of springbok, impala and blesbok sperm. Springbok, impala and blesbok post-thawed epididymal spermatozoa are capable of fertilizing domestic cow oocytes under conditions that support domestic cattle IVF. Penetration, male pronuclear formation and embryo cleavage did not differ (p>0.05) between cow oocytes inseminated with sperm from springbok, impala or blesbok however these parameters were higher (p<0.05) for oocytes inseminated with bull sperm. Modified Tyrode Lactate IVF media supported homologous fertilization and embryo development in springbok and blesbok however did not support blastocyst development. These findings suggest that cattle provide a useful model for evaluating springbok, impala and blesbok post-thawed cauda epididymal sperm functionality. Domestic cattle embryo culture conditions need to be modified to promote blastosyst development in these antelope species. Such research provides an important tool in assisted reproductive technology development when high biological value

  5. Cryosurvival and in vitro fertilizing capacity postthaw is improved when boar spermatozoa are frozen in the presence of seminal plasma from good freezer boars.

    PubMed

    Hernández, Marta; Roca, Jordi; Calvete, Juan J; Sanz, Libia; Muiño-Blanco, Teresa; Cebrián-Pérez, José A; Vázquez, Juan M; Martínez, Emilio A

    2007-01-01

    The study evaluated the protective effect of seminal plasma (SP) added to freezing extender against cryopreservation injuries to boar spermatozoa. Pooled sperm-rich fractions collected from 9 fertile boars were frozen in 0.5-mL straws after being extended in a conventional freezing extender either alone or supplemented with 5% of SPs (SP1-SP4) collected from the sperm-rich fractions (diluted 1:1, vol/vol, in Beltsville Thawing Solution extender) from 4 boars (1-4) with known sperm cryosurvival (poor, moderate, and good sperm freezers). Cryopreservation injuries were assessed in terms of postthaw sperm motility (assessed by computer-assisted sperm analysis), viability (plasma membrane and acrosome integrity assessed simultaneously by flow cytometry), membrane lipid peroxidation (malondialdehyde [MDA] production), and the ability of thawed spermatozoa to fertilize in vitro-matured homologous oocytes. The addition of SP from good sperm freezers (SP3 and SP4) improved (P < .01) the motility and viability of thawed spermatozoa without any influence on MDA production. Moreover, SP from good sperm freezers also increased (P < .05) the percentage of penetrated (SP3) and polyspermic oocytes (SP4) with respect to the control. Neither the total amount of SP proteins, protein profiles, nor antioxidant capacity of the different SPs were related to the various cryosurvival/fertilizing capacities of the processed spermatozoa. PMID:17460094

  6. Cryosurvival and in vitro fertilizing capacity postthaw is improved when boar spermatozoa are frozen in the presence of seminal plasma from good freezer boars.

    PubMed

    Hernández, Marta; Roca, Jordi; Calvete, Juan J; Sanz, Libia; Muiño-Blanco, Teresa; Cebrián-Pérez, José A; Vázquez, Juan M; Martínez, Emilio A

    2007-01-01

    The study evaluated the protective effect of seminal plasma (SP) added to freezing extender against cryopreservation injuries to boar spermatozoa. Pooled sperm-rich fractions collected from 9 fertile boars were frozen in 0.5-mL straws after being extended in a conventional freezing extender either alone or supplemented with 5% of SPs (SP1-SP4) collected from the sperm-rich fractions (diluted 1:1, vol/vol, in Beltsville Thawing Solution extender) from 4 boars (1-4) with known sperm cryosurvival (poor, moderate, and good sperm freezers). Cryopreservation injuries were assessed in terms of postthaw sperm motility (assessed by computer-assisted sperm analysis), viability (plasma membrane and acrosome integrity assessed simultaneously by flow cytometry), membrane lipid peroxidation (malondialdehyde [MDA] production), and the ability of thawed spermatozoa to fertilize in vitro-matured homologous oocytes. The addition of SP from good sperm freezers (SP3 and SP4) improved (P < .01) the motility and viability of thawed spermatozoa without any influence on MDA production. Moreover, SP from good sperm freezers also increased (P < .05) the percentage of penetrated (SP3) and polyspermic oocytes (SP4) with respect to the control. Neither the total amount of SP proteins, protein profiles, nor antioxidant capacity of the different SPs were related to the various cryosurvival/fertilizing capacities of the processed spermatozoa.

  7. [Fertility decline in Spain].

    PubMed

    Arango, J

    1987-01-01

    The historical processes of secular fertility decline in Spain and Portugal are not well understood. Very few microdemographic studies of small geographic regions or particular social strata have been done. A contribution by David Reher to the First Spanish-Portuguese-Italian Historical Demography Conference on the fertility decline in the interior province of Cuenca, Spain, uses the own-children method to analyze changes in marital fertility in the 19th and 20th centuries. Reher discovered a slight fertility decline of perhaps 15% which occurred between the end of the 18th century and 1860-75. The fertility decline did not resume until after the Spanish Civil War, and then it was a very gradual and continuous process. When instead of the total female population, women aged 35-39 were studied, unequivocal signs of fertility control appeared. Conscious fertility control thus appears to have begun among older women limiting rather than spacing births. Reher's analysis by social groups demonstrates that fertility declined first and more rapidly in the nonagricultural and urban populations and among the higher income groups. The fertility decline in Cuenca was certainly not identical to that in most of Spain, but may have been fairly typical of a large part of the interior. Another contribution to the Historical Demography Conference, by Anna Cabre and Isabel Pujadas, analyzes fertility trends and cyclical fluctuations in 20th century Cataluna, arguing that they must be placed in historical perspective if recent changes are to be understood and plausible projections made. Their work demonstrates the value of selecting a relatively homogeneous geographic unit for analysis. The contribution of Margarita Delgado to the conference analyzed interregional fertility differences in contemporary Spain. The high legitimate fertility of the south of Spain is accentuated by high nuptiality rates. In central Spain, the combination of high legitimate fertility rates and low

  8. The presence of HBV mRNA in the fertilized in vitro embryo of HBV patients confirms vertical transmission of HBV via the ovum.

    PubMed

    Ye, F; Jin, Y; Kong, Y; Shi, J Z; Qiu, H T; Zhang, X; Zhang, S L; Lin, S M

    2013-05-01

    This study aimed to confirm that vertical transmission of hepatitis B virus (HBV) can occur via the infected ovum. Specimens studied were obtained from discarded test-tube embryos from mothers with chronic HBV infection who had received in vitro fertilization treatment. Single-cell reverse transcriptase-polymerase chain reaction was used to detect HBV mRNA in the embryos. HBV mRNA was detected in the cleavage embryos of patients with chronic HBV infection, with a detection rate of 13.2% (5/38). The level of serum HBV DNA was not related to the HBV mRNA positivity rates in embryos. In this study, HBV mRNA was detected in test-tube embryos from HBV-infected mothers who had received in vitro fertilization treatment. This confirms the theory of vertical transmission of HBV via the ovum, thereby providing an important theoretical basis for further study on the mechanism of HBV vertical transmission, influencing factors and blocking measures.

  9. [[Fertility in Bangladesh villages

    PubMed

    Tsutsui, T; Igarashi, T; Tsubouchi, Y

    1990-12-01

    The authors analyze the high fertility in two Bangladesh villages based on survey and genealogical data. "Compared with nation-wide data, [the] two villages showed comparatively high fertility. There was a difference in fertility between the two villages, which appeared to result from a difference in mean age at first marriage. No other factors were identified.... As for the government's family planning program, neither village showed much effect." (SUMMARY IN ENG)

  10. In vitro maturation, fertilization, embryo development & clinical outcome of human metaphase-I oocytes retrieved from stimulated intracytoplasmic sperm injection cycles

    PubMed Central

    Álvarez, Cristina; García-Garrido, Carmen; Taronger, Roser; de Merlo, Gaspar González

    2013-01-01

    Background & objectives: The major cause of fertilisation failure after ICSI is failure of the oocyte to initiate the biochemical processes necessary for activation. This inability could be ascribed to cytoplasmic immaturity of those gametes even if they had reached nuclear maturity. The activation of a mature oocyte is characterised by release from metaphase II (MII) arrest and extrusion of the second polar body, followed by pro-nuclear formation. The aim of this study was to evaluate the fate of in vitro matured (IVM) metaphase I (MI) oocytes subjected to intracytoplasmic sperm injection (ICSI) at different time intervals after extrusion of the first polar body (1PB) in in vitro fertilization (IVF) cycles. Methods: A total of 8030 oocytes were collected from 1400 ICSI cycles, 5504 MII at the time of cumulus retrieval. Four hundred eight metaphase II (MII) (27.1%) matured to MII after in vitro culture for 2-26 h and 5389 sibling MII in the moment of oocyte denudation were injected. On the other hand, 49 ICSI cycles containing only MI oocytes at retrieval were injected at three different time intervals after reaching the MII. The intervals were as follows: 2-6 h (n=10), 8-11 h (n=4) and 23-26 h (n=10). Fertilization and development potential were evaluated in both studies. Results: Fertilization, embryo cleavage and quality were significantly lower in IVM MI compared to MII at time of denudation. Pregnancy rate was higher in group MII. Pregnancy was achieved in three embryo transfers when ICSI was performed within 2-6 h (group I) and 8-11 h (group II) after PB extrusion. One pregnancy was obtained in group I and a healthy neonate was born. Interpretation & conclusions: Immature oocytes from women whose ovaries have been stimulated could be matured, fertilized by ICSI, cleaved in vitro and to give rise to a live birth. However, the developmental competence of embryos derived from immature oocytes is reduced, compared with sibling in vivo matured oocytes. Further

  11. A Case of Unicornuate Uterus with Atypical Located Hyperstimulated Ovary after in Vitro Fertilization Pre-Embryo Transfer (IVF-ET)

    PubMed Central

    Angelova, Mariya Angelova; Kovachev, Emil Georgiev; Kisyov, Stefan Vasilev; Ivanova, Vilislava Robert

    2015-01-01

    The authors describe a case of a congenital Mullerian anomaly, uterus unicornis with missing right fallopian tube. An in Vitro Fertilization Pre-Embryo Transfer (IVF-ET) procedure was done and presently is known that the patient has left fallopian tube and left ovary, two kidneys, and right ovary is missing. No diagnostic laparoscopy and hysteroscopy were done, only hysterosalpingography (HSG) before the IVF procedure. Several days after the follicular puncture of the left ovary the patient was urgently admitted to the hospital for specialized gynaecology in Varna. Transabdominal ultrasonography showed right ovary atypically located immediately next to the liver and with emerging theca-lutein cysts. PMID:27275261

  12. Effect of different thawing temperatures on the viability, in vitro fertilizing capacity and chromatin condensation of frozen boar semen packaged in 5 ml straws.

    PubMed

    Córdova-Izquierdo, A; Oliva, J H; Lleó, B; García-Artiga, C; Corcuera, B D; Pérez-Gutiérrez, J F

    2006-03-01

    The effect of two different thawing temperatures on frozen boar semen viability, in vitro fertilizing capacity and chromatin condensation and stability was studied. Freeze-thaw motility, normal apical ridge (NAR), in vitro fertilizing (IVF) capacity and chromatin condensation and stability were evaluated after thawing at 42 degrees C, 40s and 50 degrees C, 40s. Chromatin condensation degree was determined by flow cytometry, using propidium iodide as fluorochrome intercalating agent, and chromatin stability was evaluated by the same procedure after inducing sperm chromatin decondensation with ethylene diamine tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS). The results showed that thawing straws at 42 degrees C, 40s significantly reduced motility compared to straws thawed at 50 degrees C, 40s. NAR, penetration, monospermy and polyspermy were not different between the two groups of samples thawed at different temperatures. Chromatin was significantly more compact when thawing was performed at 50 degrees C, but its stability did not show any difference relative to thawing at 42 degrees C. It is suggested that the interactions involved in chromatin overcondensation had a non-covalent nature. PMID:15975744

  13. Antagonism at combined effects of chemical fertilizers and carbamate insecticides on the rice-field N2-fixing cyanobacterium Cylindrospermum sp. in vitro

    PubMed Central

    Nayak, Nabakishore; Rath, Shakti

    2014-01-01

    Effects of chemical fertilizers (urea, super phosphate and potash) on toxicities of two carbamate insecticides, carbaryl and carbofuran, individually to the N2-fixing cyanobacterium, Cylindrospermum sp. were studied in vitro at partially lethal levels (below highest permissive concentrations) of each insecticide. The average number of vegetative cells between two polar heterocysts was 16.3 in control cultures, while the mean value of filament length increased in the presence of chemical fertilizers, individually. Urea at the 10 ppm level was growth stimulatory and at the 50 ppm level it was growth inhibitory in control cultures, while at 100 ppm it was antagonistic, i.e. toxicity-enhancing along with carbaryl, individually to the cyanobacterium, antagonism was recorded. Urea at 50 ppm had toxicity reducing effect with carbaryl or carbofuran. At 100 and 250 ppm carbofuran levels, 50 ppm urea only had a progressive growth enhancing effect, which was marked well at 250 ppm carbofuran level, a situation of synergism. Super phosphate at the 10 ppm level only was growth promoting in control cultures, but it was antagonistic at its higher levels (50 and 100 ppm) along with both insecticides, individually. Potash (100, 200, 300 and 400 ppm) reduced toxicity due to carbaryl 20 and carbofuran 250 ppm levels, but potash was antagonistic at the other insecticide levels. The data clearly showed that the chemical fertilizers used were antagonistic with both the insecticides during toxicity to Cylindrospermum sp. PMID:26038669

  14. Ovarian stimulation with gonadotropin-releasing hormone (GnRH) analogue improves the in vitro fertilization (IVF) pregnancy rate with both transvaginal and laparoscopic oocyte recovery.

    PubMed

    Dor, J; Ben-Shlomo, I; Lipitz, S; Levran, D; Etchin, A; Rudak, E; Mashiach, S

    1990-12-01

    The relative impact of ovarian stimulation protocol and oocyte retrieval technique on success rates of in vitro fertilization program was studied in 200 patients. Sixty-three patients received gonadotropin-releasing hormone analogue (GnRHa) with human menopausal gonadotropin (hMG), and 137 received hMG only. The GnRHa + hMG protocol resulted in higher pregnancy rates than the hMG-only protocol (19.0 vs 9.5%, respectively; P less than 0.01) despite a lower cleavage rate. Oocyte retrieval was performed via laparoscopy in 100 patients and transvaginally in 100 patients. The number of oocytes recovered per cycle was 6.1 +/- 3.9 with laparoscopy and 7.0 +/- 3.1 transvaginally. Pregnancy rates were similar for both retrieval techniques (13 and 12%, respectively). A breakdown of these results showed that the advantage for the GnRHa + hMG protocol was not affected by the oocyte retrieval technique. A comparison of simultaneous blood and follicular fluid pH measured every 10 min during laparoscopy and transvaginal oocyte recovery revealed a constant decline in follicular fluid pH during laparoscopy, while no changes were observed during the vaginal procedure. We conclude that the improvement in in vitro fertilization results during the period of our study is due primarily to the introduction of GnRHa + hMG protocol rather than the method of oocyte retrieval.

  15. Is the sperm-binding capability of the zona pellucida linked to its surface structure? A scanning electron microscopic study of human in vitro fertilization.

    PubMed

    Familiari, G; Nottola, S A; Micara, G; Aragona, C; Motta, P M

    1988-06-01

    The structure of the zona pellucida and the early interactions between human oocytes and spermatozoa were investigated in an in vitro fertilization program. Thirty-five mature (preovulatory) oocytes, 10 immature oocytes lacking a germinal vesicle, and 11 atretic oocytes which had not undergone fertilization at 10-20 hr after insemination were studied by light and scanning electron microscopy. Observed through employment of these techniques, the zona pellucida showed two basically different patterns: a mesh-like, spongy structure having wide and/or close meshes; and a compact, smooth surface. The smooth-surfaced zona was most commonly seen in the cultured oocytes belonging to the immature and atretic groups. These observations seem to show that the spongy appearance of the zona pellucida is related mainly to oocyte development and maturity. In this study, greater numbers of penetrating spermatozoa were noted on oocytes showing the mesh-like zona, in contrast to the presence of a few sperm flattened against its surface or the frank absence of sperm associated with oocytes having the more compact, smooth zona. It is likely that the condensation of the outer aspect of the zona pellucida causes a disorientation of sperm-binding sites, which would probably result in markedly reduced binding and penetration capacity with spermatozoa. These changes might ultimately lead to impairment of in vitro oocyte fertilizability.

  16. Mouse early embryos obtained by natural breeding or in vitro fertilization display a differential sensitivity to extremely low-frequency electromagnetic fields.

    PubMed

    Beraldi, Rosanna; Sciamanna, Ilaria; Mangiacasale, Rosamaria; Lorenzini, Rodolfo; Spadafora, Corrado

    2003-07-01

    We have investigated the sensitivity of pre-implantation embryos obtained by natural breeding (NB) or in vitro fertilization (IVF) to extremely low-frequency magnetic fields (ELF-MF). Fertilized eggs obtained by NB were removed from mothers 12h after mating and cultured in vitro for 5 days under continuous ELF-MF exposure (constant strength of 50Hz and various intensities, i.e. 60, 120 and 220 microT). Alternatively, zygotes obtained by IVF were subjected to ELF-MF exposure (50Hz, 60 microT), starting 12h after IVF for 5 days. We found that ELF-MF exposure causes a small yet significant (P<0.05) decrease in the survival rate of NB-derived embryos at the latest stages of pre-implantation development, i.e. the eight cell-to-blastocyst transition. In embryos exposed to the highest field intensity (220 microT), the effect became apparent somewhat earlier. When IVF-derived embryos were exposed to ELF-MF, the reduction in the rate of embryo survival was more pronounced and the difference from controls was more significant (P<0.01). Moreover, the decreased survival rate in IVF embryos became apparent as early as the first cleavage and persisted throughout pre-implantation. These results suggest that IVF-derived embryos are more sensitive than NB-generated embryos to ELF-MF, and that this sensitivity occurs earlier in development.

  17. Fertility decline in Paraguay.

    PubMed

    Ishida, Kanako; Stupp, Paul; Melian, Mercedes

    2009-09-01

    Recent reproductive health surveys show that the fertility rate in Paraguay decreased precipitously from 4.3 lifetime births per woman in 1995-98 to 2.9 births in 2001-04. In this study, we establish data consistency between the 1998 and 2004 surveys by comparing a series of cohort-specific period rates and use the Bongaarts framework of proximate determinants of fertility to demonstrate that an increase in the contraceptive prevalence rate (CPR) between 1998 and 2004 fully accounts for the fertility decline. Decomposition of rates shows that changes in group-specific CPRs explain a greater proportion of the change in the overall CPR than do changes in population composition by educational attainment, urban residence, region, and language spoken at home. Finally, we show that younger cohorts of women in 2004 reported ideal completed fertility desires of less than 2.9 births, suggesting that the fertility rate is likely to continue to decrease.

  18. Effect of nitrogen fixation, nitrogen fertilization and viral infection on yield, tannin and protein contents and in vitro protein digestibility of faba bean.

    PubMed

    Babiker, E E; el Sheikh, E A; Osman, A J; el Tinay, A H

    1995-04-01

    A field investigation of two faba bean cultivars (cv.), Agabat and Silaim, showed that bean yellow mosaic virus (BYMV) infection reduced (p < or = 0.001) yield (Kg/ha), protein content and in vitro protein digestibility (IVPD) but increased (p < or = 0.05) tannin content (mg/100 ml). Nitrogen fertilization with viral infection significantly reduced yield and IVPD for cv. Silaim and increased (p < or = 0.05) protein and tannin contents. Nitrogen fertilization alone was found to increase (p < or = 0.05) yield, protein and tannin contents but slightly reduced IVPD. Rhizobium inoculation with viral infection significantly decreased yield per unit area, protein content and IVPD, but increased (p < or = 0.05) tannin content. Rhizobium inoculation alone significantly increased (p < or = 0.001) yield and tannin content and slightly increased protein content but decreased IVPD. The results indicated that nitrogen fertilization or nitrogen fixation increased yield, protein and tannin contents and decreased IVPD. Viral infection had an adverse effect on yield, protein content and IVPD but had no effect on tannin content.

  19. A formula for scoring human embryo growth rates in in vitro fertilization: its value in predicting pregnancy and in comparison with visual estimates of embryo quality.

    PubMed

    Cummins, J M; Breen, T M; Harrison, K L; Shaw, J M; Wilson, L M; Hennessey, J F

    1986-10-01

    Two systems for measuring embryo development in vitro were evaluated. One was a 1-4 scale based on a subjective evaluation of embryo quality (EQ) from microscopic appearance. In addition, a formula for scoring embryo growth rate in vitro was developed. The embryo development rating (EDR) was based on the ratio between the time at which embryos were observed at a particular stage after insemination and the time at which they would be expected to reach that stage in a hypothetical "ideal" growth rate with a cell cycle length of 11.9 hr. Using this scoring system, "normally" growing embryos scored 100. This approach was aimed at partially normalizing the data and allowed all embryos to be analyzed similarly regardless of the time of observation. Analysis of 1539 embryo replacements resulting in 232 clinical pregnancies showed that both EDR and embryo-quality scores were of value in predicting success, with clinical pregnancy most likely to eventuate from a combination of moderate to good EQ scores (2-4) coupled with average or above-average growth rates (EDR scores from 90 to 129). Poor-quality and very slowly or very rapidly growing embryos were underrepresented in cycles that proceeded to pregnancy. These inferences were based on all embryos transferred (mean, 2.73 per transfer cycle), and they were substantiated by an analysis of 33 pregnancies resulting from replacement of a single embryo and from 18 pregnancies in which all embryos scored the same with both systems. EQ and EDR were significantly associated with each other and together provide a valuable guide in predicting pregnancy, in selecting embryos for freezing, and in monitoring day-to-day performance in the in vitro fertilization (IVF) program.

  20. In vitro maturation and fertilization of oocytes recovered from free-ranging Burchell's zebra (Equus burchelli) and Hartmann's zebra (Equus zebra hartmannae).

    PubMed

    Meintjes, M; Bezuidenhout, C; Bartels, P; Visser, D S; Meintjes, J; Loskutoff, N M; Fourie, F L; Barry, D M; Godke, R A

    1997-09-01

    A noninvasive repeatable method to harvest oocytes for in vitro fertilization (IVF) could potentially be used to assist reproduction in endangered equid species. The objectives of this study were to evaluate a specific transvaginal ultrasound-guided oocyte recovery procedure for use in zebra mares and the general applicability of IVF procedures in zebra. Ovaries were collected from Burchell's zebra (Equus burchelli) and Hartmann's zebra (Equus zebra hartmannae) mares at routine culling for Expt. I. Of the 144 oocytes recovered from these ovaries, 70% were of excellent quality. No significant difference in oocyte quality was found between the two zebra species. Zona drilling was performed on in vitro-matured oocytes prior to IVF. Epididymal sperm from culled Burchell's zebra stallions were used for IVF. The sperm either were exposed to calcium ionophore or were not treated and served as a control. In vitro fertilized oocytes were then co-cultured with zebra granulosa cells (ZGC) or with bovine oviduct cells (BOC) for up to 8 days. Overall, a 38% cleavage rate was obtained with 16% of sperm-exposed oocytes developing to the morula or blastocyst stage. All of the embryos that developed to at least the morula stage were cultured on BOC; whereas, none of those cultured on ZGC reached the morula stage during the same interval. Cleavage rates of oocytes inseminated with ionophore-treated or with control sperm were not significantly different, suggesting that ionophore treatment of epididymal sperm for IVF in these zebra species may be redundant. In Expt. II, 10 transvaginal ultrasound-guided oocyte aspiration procedures on five captive Burchell's zebra mares recovered a total of 33 oocytes (6.6 oocytes/female) of which 94% were considered viable. This approach may be an attractive means of producing gametes for assisted reproduction in endangered species. Furthermore, results from this study indicate that IVF may become a means of producing offspring from zebra and other

  1. 22q11.2 Deletion Syndrome due to a Translocation t(6;22) in a Patient Conceived via in vitro Fertilization

    PubMed Central

    Gollo Dantas, Anelisa; Bortolai, Adriana; Moysés-Oliveira, Mariana; Takeno Herrero, Sylvia; Azoubel Antunes, Adriana; Tavares Costa-Carvalho, Beatriz; Ayres Meloni, Vera; Melaragno, Maria Isabel

    2016-01-01

    We report on a patient conceived via in vitro fertilization (IVF) with a 22q11.2 deletion due to an unusual unbalanced translocation involving chromosomes 6 and 22 in a karyotype with 45 chromosomes. Cytogenomic studies showed that the patient has a 3.3-Mb deletion of chromosome 22q and a 0.4-Mb deletion of chromosome 6p, which resulted in haploinsufficiency of the genes responsible for the 22q11.2 deletion syndrome and also of the IRF4 gene, a member of the interferon regulatory factor family of transcription factors, which is expressed in the immune system cells. The rearrangement could be due to the manipulation of the embryo or as a sporadic event unrelated to IVF. Translocation involving chromosome 22 in a karyotype with 45 chromosomes is a rare event, with no previous reports involving chromosomes 6p and 22q. PMID:26997945

  2. Reproductive Performance of Holstein Dairy Cows Grazing in Dry-summer Subtropical Climatic Conditions: Effect of Heat Stress and Heat Shock on Meiotic Competence and In vitro Fertilization.

    PubMed

    Pavani, Krishna; Carvalhais, Isabel; Faheem, Marwa; Chaveiro, Antonio; Reis, Francisco Vieira; da Silva, Fernando Moreira

    2015-03-01

    The present study was designed to evaluate how environmental factors in a dry-summer subtropical climate in Terceira-Azores (situated in the North Atlantic Ocean: 38° 43' N 27° 12' W) can affect dairy cow (Holstein) fertility, as well as seasonal influence on in vitro oocytes maturation and embryos development. Impact of heat shock (HS) effects on in vitro oocyte's maturation and further embryo development after in vitro fertilization (IVF) was also evaluated. For such purpose the result of the first artificial insemination (AI) performed 60 to 90 days after calving of 6,300 cows were recorded for one year. In parallel, climatic data was obtained at different elevation points (n = 5) from 0 to 1,000 m and grazing points from 0 to 500 m, in Terceira island, and the temperature humidity index (THI) was calculated. For in vitro experiments, oocytes (n = 706) were collected weekly during all year, for meiotic maturation and IVF. Further, to evaluate HS effect, 891 oocytes were collected in the cold moths (December, January, February and March) and divided in three groups treated to HS for 24 h during in vitro maturation at: C (Control = 38.5°C), HS1 (39.5°C) and HS2 (40.5°C). Oocytes from each group were used for meiotic assessment and IVF. Cleavage, morula and blastocyst development were evaluated respectively on day 2, 6, and 9 after IVF. A negative correlation between cow's conception rate (CR) and THI in grazing points (-91.3%; p<0.001) was observed. Mean THI in warmer months (June, July, August and September) was 71.7±0.7 and the CR (40.2±1.5%) while in cold months THI was 62.8±0.2 and CR was 63.8±0.4%. A similar impact was obtained with in vitro results in which nuclear maturation rate (NMR) ranged from 78.4% (±8.0) to 44.3% (±8.1), while embryos development ranged from 53.8% (±5.8) to 36.3% (±3.3) in cold and warmer months respectively. In vitro HS results showed a significant decline (p<0.05) on NMR of oocytes for every 1°C rising temperature (78

  3. Reproductive Performance of Holstein Dairy Cows Grazing in Dry-summer Subtropical Climatic Conditions: Effect of Heat Stress and Heat Shock on Meiotic Competence and In vitro Fertilization

    PubMed Central

    Pavani, Krishna; Carvalhais, Isabel; Faheem, Marwa; Chaveiro, Antonio; Reis, Francisco Vieira; da Silva, Fernando Moreira

    2015-01-01

    The present study was designed to evaluate how environmental factors in a dry-summer subtropical climate in Terceira-Azores (situated in the North Atlantic Ocean: 38° 43′ N 27° 12′ W) can affect dairy cow (Holstein) fertility, as well as seasonal influence on in vitro oocytes maturation and embryos development. Impact of heat shock (HS) effects on in vitro oocyte’s maturation and further embryo development after in vitro fertilization (IVF) was also evaluated. For such purpose the result of the first artificial insemination (AI) performed 60 to 90 days after calving of 6,300 cows were recorded for one year. In parallel, climatic data was obtained at different elevation points (n = 5) from 0 to 1,000 m and grazing points from 0 to 500 m, in Terceira island, and the temperature humidity index (THI) was calculated. For in vitro experiments, oocytes (n = 706) were collected weekly during all year, for meiotic maturation and IVF. Further, to evaluate HS effect, 891 oocytes were collected in the cold moths (December, January, February and March) and divided in three groups treated to HS for 24 h during in vitro maturation at: C (Control = 38.5°C), HS1 (39.5°C) and HS2 (40.5°C). Oocytes from each group were used for meiotic assessment and IVF. Cleavage, morula and blastocyst development were evaluated respectively on day 2, 6, and 9 after IVF. A negative correlation between cow’s conception rate (CR) and THI in grazing points (−91.3%; p<0.001) was observed. Mean THI in warmer months (June, July, August and September) was 71.7±0.7 and the CR (40.2±1.5%) while in cold months THI was 62.8±0.2 and CR was 63.8±0.4%. A similar impact was obtained with in vitro results in which nuclear maturation rate (NMR) ranged from 78.4% (±8.0) to 44.3% (±8.1), while embryos development ranged from 53.8% (±5.8) to 36.3% (±3.3) in cold and warmer months respectively. In vitro HS results showed a significant decline (p<0.05) on NMR of oocytes for every 1°C rising

  4. Maintenance of meiotic arrest in bovine oocytes using the S-enantiomer of roscovitine: effects on maturation, fertilization and subsequent embryo development in vitro.

    PubMed

    Coy, Pilar; Romar, Raquel; Payton, Rebecca R; McCann, Lisa; Saxton, Arnold M; Edwards, J Lannett

    2005-01-01

    The overall objective was to evaluate the effectiveness of the S-enantiomer of roscovitine (inhibitor of p34cdc2/cyclin B kinase) to maintain bovine cumulus-oocyte complexes at the germinal vesicle (GV) stage for extended times after removal from antral follicles without compromising subsequent maturation, fertilization and embryo development. Oocytes were cultured in 0, 12.5, 25 or 50 micromol/l S-roscovitine for 24 h. Hoechst staining showed that 50 micromol/l S-roscovitine maintained >90% of oocytes at the GV stage and inhibited gonadotropin-induced cumulus expansion. Fewer oocytes underwent nuclear maturation after in vitro maturation (Hoechst staining) when cultured in 50 micromol/l S-roscovitine for 66 versus 21 or 42 h. Zona pellucida (ZP) hardening (pronase resistance), cortical granule types (lens culinaris agglutinin-fluorescein isothiocyanate), nuclear maturation and fertilization with frozen-thawed spermatozoa (Hoechst staining) were assessed after culture of oocytes in 50 micromol/l S-roscovitine for 0, 24 or 48 h. Neither ZP hardening, nor nuclear maturation nor fertilization were altered by roscovitine culture for 48 h. A higher proportion of oocytes had a type III cortical granule pattern (premature translocation to the oolemma) after roscovitine culture for 48 h. However, embryo development was not compromised as cleavage, development to 8-16 cell and blastocyst stages were at least comparable in control and roscovitine-treated oocytes. In conclusion, the studies have shown that S-roscovitine reversibly maintained bovine oocytes at the GV stage for 48 h. However, maintenance of oocytes in static culture for 48 h was not sufficient to improve development above non-treated controls. PMID:15615895

  5. Improved embryo development in Japanese black cattle by in vitro fertilization using ovum pick-up plus intracytoplasmic sperm injection with dithiothreitol.

    PubMed

    Oikawa, Toshinori; Itahashi, Tomoko; Numabe, Takashi

    2016-01-01

    The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse, ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with low embryo production compared with the standard OPU-IVF.

  6. Improved embryo development in Japanese black cattle by in vitro fertilization using ovum pick-up plus intracytoplasmic sperm injection with dithiothreitol

    PubMed Central

    OIKAWA, Toshinori; ITAHASHI, Tomoko; NUMABE, Takashi

    2015-01-01

    The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse, ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with low embryo production compared with the standard OPU-IVF. PMID:26460690

  7. Anti-mullerian hormone cut-off values for predicting poor ovarian response to exogenous ovarian stimulation in in-vitro fertilization

    PubMed Central

    Satwik, Ruma; Kochhar, Mohinder; Gupta, Shweta M; Majumdar, Abha

    2012-01-01

    OBJECTIVES: (a) To establish the cut-off levels for anti-Mullerian hormone (AMH) in a population of Indian women that would determine poor response. (b) To determine which among the three ie.,: age, follicle stimulating hormone (FSH), or AMH, is the better determinant of ovarian reserve. STUDY DESIGN: Prospective observational study. SETTING: In vitro fertilization (IVF) unit of a tertiary hospital. MATERIALS AND METHODS: The inclusion criterion was all women who presented to the center for in-vitro fertilization/Intracytoplasmic sperm injection (IVF/ICSI). The exclusion criteria were age >45 years, major medical illnesses precluding IVF or pregnancy, FSH more than 20 IU/L, and failure to obtain consent. The interventions including baseline pelvic scan, day 2/3 FSH, luteinizing hormone (LH), estradiol estimations, and AMH measurement on any random day of cycle were done. Subjects underwent IVF according to long agonist or antagonist protocol regimen. Oocyte recovery was correlated with studied variables. The primary outcome measure was the number of oocytes aspirated (OCR). Three categories of ovarian response were defined: poor response, OCR ≤ 3; average response, OCR between 4 and 15; hyperresponse, OCR > 15. RESULTS: Of the 198 patients enrolled, poor, average, and hyperresponse were observed in 23%, 63%, and 14% respectively. Correlation coefficient for AMH with ovarian response was r = 0.591. Area under the curve (AUCs) for poor response for AMH, subject's age, and FSH were 0.768, 0.624, and 0.635, respectively. The discriminatory level of AMH for prediction of absolute poor response was 2 pmoL/l, with 98% specificity and 20% sensitivity. CONCLUSIONS: AMH fares better than age and FSH in predicting the overall ovarian response and poor response, though it cannot be the absolute predictor of non-responder status. A level of 2 pmol/l is discriminatory for poor response. PMID:23162361

  8. Improvement in in vitro fertilization rate, decrease in reactive oxygen species and spermatozoa death incidence in rams by dietary fish oil.

    PubMed

    Matini Behzad, A; Ebrahimi, B; Alizadeh, A R; Esmaeili, V; Dalman, A; Rashki, L; Shahverdi, A H

    2014-08-01

    Our aim was to evaluate the effects of fish oil feeding on sperm classical parameters, level of reactive oxygen spices (ROS), spermatozoa death incidence and in vitro fertilization (IVF) rate in rams. We randomly assigned nine rams, into two experimental groups (isoenergetic and isonitrogenous rations with constant level of vitamin E supplement): control (CTR; n = 5) and fish oil (FO; n = 4, 35 g/day/ram). Diets were fed for 70 days during the physiological breeding season. After a 21-day dietary adaptation period, semen was collected weekly from each ram by an artificial vagina. Sperm classical parameters were determined by the computer-assisted sperm analyzer system (CASA), and it was prepared for IVF process by swim-up technique. These evaluations were performed during the first and last weeks of sampling. Intracellular ROS level and spermatozoa death incidence were detected by flow cytometry on a weekly basis after adaptation. Data were analysed with SPSS 15. The volume, concentration (3.6 and 2.7 × 10(9) /ml) and sperm progressive motility (60 and 48%) were significantly improved in the FO group compared with the CTR (p < 0.05). A comparison of two-cell stage embryos following IVF in the two groups showed a significantly higher fertilization rate in the FO group (56%) compared with the CTR (49%). Superoxide anion (O2 (-) ) rate was significantly lower (p < 0.05) at the third week of sampling in the FO. Although the H2 O2 rate was numerically lower in the FO group compared with the CTR, this difference was not significant. In addition, apoptosis showed a significant difference in the third week of sampling (15 and 30% for FO and CTR, respectively; p < 0.05). Overall, adding fish oil to the ram diet not only improved sperm quality and IVF results, it also could reduce oxygen-free radicals and the incidence of spermatozoa death.

  9. Creating the ‘ethics industry': Mary Warnock, in vitro fertilization and the history of bioethics in Britain

    PubMed Central

    Wilson, Duncan

    2011-01-01

    Recent decades have seen a shift in the management and discussion of biomedicine. Issues once considered by doctors and scientists are now handled by a diverse array of participants, including philosophers, lawyers, theologians and lay representatives. This new approach, known as ‘bioethics', has become the norm in regulatory committees and public debate. In this article, I argue that bioethics emerged as a valued enterprise in Britain during the 1980s because it fulfilled, and linked, the concerns of several groups. My analysis centres on the moral philosopher Mary Warnock, who chaired a government inquiry into human fertilization and embryology between 1982 and 1984, and became a strong advocate of bioethics. I detail how Warnock's promotion of bioethics tallied with the Conservative government's desire for increased surveillance of hitherto autonomous professions – while fulfilling her own belief that philosophers should engage in public affairs. And I also show that Warnock simultaneously promoted bioethics to doctors and scientists as an essential safeguard against declining political and public trust. This stance, I argue, framed bioethics as a vital intermediary between politics, the public, and biomedicine, and explains the growth and endurance of what the Guardian identified as an ethics industry. PMID:22563348

  10. Superovulation of mice with human menopausal gonadotropin or pure follicle-stimulating hormone in combination with human chorionic gonadotropin and the effects of oocyte aging on in vitro fertilization.

    PubMed

    Edirisinghe, W R; Law, H Y; NG, S C; Chia, C M; Ratnam, S S

    1986-10-01

    The response of female mice of F1 hybrids (CBA x C57/BL) to superovulatory doses of human menopausal gonadotropin (hMG) or pure follicle-stimulating hormone (FSH) in combination with human chorionic gonadotropin (hCG) was studied. Furthermore, the effect of oocyte aging in vivo on the subsequent rate of fertilization in vitro was also investigated. The oocytes were collected at 12, 18, and 24 hr after hCG injection and in vitro fertilization (IVF) was carried out in T6 medium. A higher proportion of animals responded to hMG stimulation (32/70) compared to pure FSH (15/66). Furthermore, hMG gave a higher oocyte recovery (454/32) than pure FSH (77/15). Fertilization rates of 57.8, 51.5, and 53.5% were obtained for the 12-, 18-, and 24-hr groups, respectively, after correction for parthenogenetic division of oocytes in the controls. No significant differences in fertilization rates were observed among the three time intervals used in recovering oocytes. However, as the degeneration and parthenogenetic division increased with the delay in collection of oocytes, 12 hr post-hCG injection was the best time to collect oocytes to obtain optimum results in in vitro fertilization.

  11. Effect of growth differentiation factor‑9 C447T and G546A polymorphisms on the outcomes of in vitro fertilization.

    PubMed

    Serdyńska-Szuster, Monika; Jędrzejczak, Piotr; Ożegowska, Katarzyna Ewa; Hołysz, Hanna; Pawelczyk, Leszek; Jagodziński, Paweł Piotr

    2016-05-01

    Single nucleotide polymorphisms (SNPs) in the growth differentiation factor (GDF)‑9 gene are associated with premature ovarian failure, insufficient ovarian stimulation and a poor in vitro fertilization (IVF) score in women with diminished ovarian reserve. The aim of the present study was to assess the effect of C447T (rs254286) and G546A (rs10491279) SNPs on ovary stimulation response, oocyte quality, fertilization rate and outcome of clinical pregnancy in an infertile population of Polish females (n=86) treated with IVF. The present study also included a group of fertile women (n=202). The P‑trend value, calculated for the GDF‑9 C447T transition in infertile women, was statistically significant and were equal to 0.0195. A significant association of the GDF‑9 C447T SNP was observed with infertility for the C/C vs. T/T + C/T model (OR= 2.140; 95% CI=1.043‑4.393; P=0.0349). The GDF‑9 G546A SNP was significantly associated with the G/A vs. G/G model with poor ovarian stimulation (OR=9.303; 95% CI=2.568‑33.745; P=0.0008) and poor fertilization rate (OR=2.981; 95% CI=1.033‑8.607; P=0.0385). For the GDF‑9 C447T SNP, a significant association was observed between the C/C + C/T vs. T/T model and a poor ovarian stimulation response (OR=15.309; 95% CI=0.875‑267.83; P=0.0078), and a poor fertilization rate (OR=4.842; 95% CI=1.310‑17.901; P=0.0121). The present genetic evaluation revealed associations between IVF outcomes and the GDF‑9 A546G and C447T SNPs. Additionally, these results indicated that the GDF‑9 C447T SNP is a possible candidate genetic risk factor for female infertility in the Polish population.

  12. The effect of extender, method of thawing, and duration of storage on in vitro fertility measures of frozen-thawed boar sperm.

    PubMed

    Knox, R V; Ringwelski, J M; McNamara, K A; Aardsma, M; Bojko, M

    2015-08-01

    Frozen-thawed boar sperm (FTS) has reduced in vitro and in vivo life span compared to liquid semen. Experiments tested whether extenders, thawing procedures, and storage temperatures could extend the fertile life span of FTS. Experiment 1 tested the effect of six extenders on postthaw motility (MOT) and viability (VIA). Straws from boars (n = 6) were thawed, diluted into each extender, and evaluated at 20, 60, and 120 minutes. There was a trend (P = 0.08) for an extender-by-time interaction for MOT and effect of extender and time for MOT (P < 0.0001) and extender (P = 0.10) and time (P < 0.0001) for VIA. Experiment 2 evaluated the effect of temperature and time of thawing on in vitro fertility at intervals after thawing. Straws (0.5 mL) from different boar ejaculates (n = 15) were thawed at 50 °C for 10, 20, or 30 seconds or at 70 °C for 5, 10, or 20 seconds and evaluated at 5, 30, and 60 minutes. There was an effect of thawing treatment on MOT, VIA, and ACR (viable sperm with intact acrosomes, P < 0.0001) and an effect of time of evaluation (P < 0.0001) on MOT and ACR. Thawing at 70 °C for 20 seconds reduced (P < 0.05) MOT, VIA, and ACR compared to other treatments. Experiment 3 tested the effects of storage temperature and time after thawing using 20 ejaculates. Samples were thawed, diluted, and allotted to storage at 17 °C, 26 °C, or 37 °C with evaluation at 2, 6, 12, and 24 hours. There was a storage temperature and time effect and an interaction for MOT and VIA (P < 0.0001). Storage at 17 °C and 26 °C increased (P < 0.05) MOT over all times (38.5%) compared to 37 °C (26%), whereas MOT was reduced at intervals. Viability was also greatest with 17 °C and 26 °C compared to 37 °C and was also affected by time and decreased with time. These results indicate that FTS can be held at 17 °C or 26 °C for up to 2 hours before use and would allow for preparation of multiple doses. These data suggest in vitro fertility of FTS is affected by extenders, thawing

  13. Comparison of microscopic epididymal sperm aspiration and intracytoplasmic sperm injection/in-vitro fertilization with repeat microscopic reconstruction following vasectomy: is second attempt vas reversal worth the effort?

    PubMed

    Donovan, J F; DiBaise, M; Sparks, A E; Kessler, J; Sandlow, J I

    1998-02-01

    Since 1986, we have performed microscopic reconstruction in 18 men following failed microscopic vasectomy reversal. Between 1994 and 1996, nine couples have undergone microscopic epididymal sperm aspiration (MESA)/ intracytoplasmic sperm injection (ICSI) treatment for male infertility due either to congenital absence of the vas deferens (CAVD) or inoperable excurrent duct obstruction. We compared the cost efficiency of repeat vasectomy reversal to that for MESA combined with ICSI/in-vitro fertilization (ICSI/IVF). The cost of male partner procedures (vasectomy reversal, MESA) was based on physician and hospital charges, while the cost of ICSI/IVF included preparation of the female partner (medications and physician charges) and procedures (physician and hospital charges including oocyte retrieval, micromanipulation, and embryo transfer). Our cost examination does not include charges related to follow-up visits, prenatal monitoring, complications of pregnancy (i.e. miscarriage) or delivery in either group. Overall patency and pregnancy rates in the repeat vasectomy reversal group were 78 and 44% respectively. The cost per delivered baby (including multiple metachronos deliveries per couple) was $14892. Fertilization of oocytes has been achieved in 37/72 (51%) and pregnancies have occurred in 6/9 (67%) attempts and 5/9 (56%) report delivery. The average cost per pregnancy was $25637 and the average cost per delivered baby (or ongoing pregnancy) was $35570. The cost per delivery by MESA/ ICSI/IVF is 2.4 times the charges per delivery obtained through repeat vasectomy repair. Couples attempting to overcome infertility caused by vasal obstruction should be informed that vas reconstruction remains a cost effective means of re-establishing fertility even in men who have previously failed vasectomy reversal.

  14. Metabolomic profiling of human follicular fluid from patients with repeated failure of in vitro fertilization using gas chromatography/mass spectrometry

    PubMed Central

    Xia, Lan; Zhao, Xiaoming; Sun, Yun; Hong, Yan; Gao, Yuping; Hu, Shuanggang

    2014-01-01

    Objective: To establish a gas chromatography/mass spectrometry (GC/MS)-based metabolomics method to compare the metabolites in the follicular fluid (FF) from patients with in vitro fertilization (IVF) and repeated IVF failure (RIF). Methods: A prospective study was employed in Center for Reprodutive Medcine, Renji Hospital, Shanghai, China, between January and October 2010. FF samples were collected from 13 patients with RIF and 15 patients who achieved pregnancy after the first IVF cycle. Results: Partial least squares (PLS) discriminant analysis of the PCA data revealed that the samples were scattered into two different regions. FF from the two groups differed with respect to 20 metabolites. FF from RIF group showed elevated levels of several amino acids (valine, threonine, isoleucine, cysteine, serine, proline, alanine, phenylalanine, lysine, methionine and ornithine), and reduced levels of dicarboxylic acids, cholesterol and some organic acids. Conclusions: The studies corroborated successful determination of the levels of metabolite in the FF. PMID:25400819

  15. Factors affecting efficiency of introducing foreign DNA and RNA into parthenogenetic or in vitro-fertilized porcine eggs by cytoplasmic microinjection.

    PubMed

    Liu, Shuai; Liu, XiaoQun; Huang, HaiYan; Liu, QingYou; Su, XiaoPing; Zhu, Peng; Li, HongLi; Cui, KuiQing; Xie, BingKun; Shi, DeShun

    2016-08-01

    Cytoplasmic microinjection (CI) of foreign gene into in vivo fertilized zygotes has emerged as a useful tool for transgenic pig production. In the current study, we investigated factors affecting transgenic efficiency and developmental potential of parthenogenetic (PA) and in vitro-fertilized (IVF) porcine embryos produced by CI. These factors included adding of RNase inhibitor, DNA or RNA concentration, injection time, and different structures of plasmids. Our results showed that adding of 1-4 U/μL of RNase inhibitor did not have negative effect on development potential of CI-PA embryos, and RNase inhibitor injection significantly increased EGFP expressing rate of CI-PA embryos. High injection DNA concentration and long injection interval after PA significantly reduced blastocyst formation. Different molecular structures such as DNA or RNA affected CI-PA embryos development, and RNA had little harmful effect on pig's early embryonic development. EGFP expression rate of CI-IVF embryos was improved following the increase of foreign DNA concentration, but blastocyst formation rate was decreased. Injection time after IVF did not show any significant difference on embryonic development, but longer interval resulted in a significantly lower EGFP expressing rate. Cas9 mRNA and myostatin (GDF-8) sgRNA co-injection indicated that the mutation rate of CI-IVF group was significantly higher than that of CI-PA. The CI-IVF-generated embryos were then transferred to six recipient pigs, but no live piglets were obtained. The following pronuclear formation assessment showed more than 76.1% IVF zygotes were polyspermy. These results demonstrate that CI-PA and CI-IVF were effective methods for production of transgenic pig embryos. However, polyspermic fertilization and poor quality of porcine IVF blastocysts are still the main problem of resulting in pregnancy failure.

  16. Factors affecting efficiency of introducing foreign DNA and RNA into parthenogenetic or in vitro-fertilized porcine eggs by cytoplasmic microinjection.

    PubMed

    Liu, Shuai; Liu, XiaoQun; Huang, HaiYan; Liu, QingYou; Su, XiaoPing; Zhu, Peng; Li, HongLi; Cui, KuiQing; Xie, BingKun; Shi, DeShun

    2016-08-01

    Cytoplasmic microinjection (CI) of foreign gene into in vivo fertilized zygotes has emerged as a useful tool for transgenic pig production. In the current study, we investigated factors affecting transgenic efficiency and developmental potential of parthenogenetic (PA) and in vitro-fertilized (IVF) porcine embryos produced by CI. These factors included adding of RNase inhibitor, DNA or RNA concentration, injection time, and different structures of plasmids. Our results showed that adding of 1-4 U/μL of RNase inhibitor did not have negative effect on development potential of CI-PA embryos, and RNase inhibitor injection significantly increased EGFP expressing rate of CI-PA embryos. High injection DNA concentration and long injection interval after PA significantly reduced blastocyst formation. Different molecular structures such as DNA or RNA affected CI-PA embryos development, and RNA had little harmful effect on pig's early embryonic development. EGFP expression rate of CI-IVF embryos was improved following the increase of foreign DNA concentration, but blastocyst formation rate was decreased. Injection time after IVF did not show any significant difference on embryonic development, but longer interval resulted in a significantly lower EGFP expressing rate. Cas9 mRNA and myostatin (GDF-8) sgRNA co-injection indicated that the mutation rate of CI-IVF group was significantly higher than that of CI-PA. The CI-IVF-generated embryos were then transferred to six recipient pigs, but no live piglets were obtained. The following pronuclear formation assessment showed more than 76.1% IVF zygotes were polyspermy. These results demonstrate that CI-PA and CI-IVF were effective methods for production of transgenic pig embryos. However, polyspermic fertilization and poor quality of porcine IVF blastocysts are still the main problem of resulting in pregnancy failure. PMID:27130683

  17. Fertility preservation in young patients with cancer

    PubMed Central

    Suhag, Virender; Sunita, B. S.; Sarin, Arti; Singh, A. K.; Dashottar, S.

    2015-01-01

    Infertility can arise as a consequence of treatment of oncological conditions. The parallel and continued improvement in both the management of oncology and fertility cases in recent times has brought to the forefront the potential for fertility preservation in patients being treated for cancer. Many survivors will maintain their reproductive potential after the successful completion of treatment for cancer. However total body irradiation, radiation to the gonads, and certain high dose chemotherapy regimens can place women at risk for acute ovarian failure or premature menopause and men at risk for temporary or permanent azoospermia. Providing information about risk of infertility and possible interventions to maintain reproductive potential are critical for the adolescent and young adult population at the time of diagnosis. There are established means of preserving fertility before cancer treatment; specifically, sperm cryopreservation for men and in vitro fertilization and embryo cryopreservation for women. Several innovative techniques are being actively investigated, including oocyte and ovarian follicle cryopreservation, ovarian tissue transplantation, and in vitro follicle maturation, which may expand the number of fertility preservation choices for young cancer patients. Fertility preservation may also require some modification of cancer therapy; thus, patients’ wishes regarding future fertility and available fertility preservation alternatives should be discussed before initiation of therapy. PMID:26942145

  18. Efficient genetic manipulation of the NOD-Rag1-/-IL2RgammaC-null mouse by combining in vitro fertilization and CRISPR/Cas9 technology.

    PubMed

    Li, Feng; Cowley, Dale O; Banner, Debra; Holle, Eric; Zhang, Liguo; Su, Lishan

    2014-06-17

    Humanized mouse models have become increasingly important and widely used in modeling human diseases in biomedical research. Immunodeficient mice such as NOD-Rag1-/-IL2RgammaC-null (NRG) or NOD-SCID-IL2RgammaC-null (NSG) mice are critical for efficient engraftment of human cells or tissues. However, their genetic modification remains challenging due to a lack of embryonic stem cells and difficulty in the collection of timed embryos after superovulation. Here, we report the generation of gene knockout NRG mice by combining in vitro fertilization (IVF) and CRISPR/Cas9 technology. Sufficient numbers of fertilized embryos were produced through IVF, and a high rate of Fah gene targeting was achieved with microinjection of Cas9 mRNA, gRNA and single strand oligonucleotide DNA (ssDNA) into the embryos. The technology paves the way to construct NRG or NSG mutant mice to facilitate new humanized mouse models. The technology can also be readily adapted to introduce mutations in other species such as swine and non-human primates.

  19. Fertilization rates and in vitro embryo production using sexed or non-sexed semen selected with a silane-coated silica colloid or Percoll.

    PubMed

    Rodríguez Villamil, P; Wei, H; Moreira, G; Caccia, M; Fernandez Taranco, M; Bó, G A

    2012-07-01

    The aim of this study was to evaluate sperm fertilization rates and in vitro embryo development rates for sexed and non-sexed semen selected using a silane-coated silica colloid method (Isolate) or Percoll. Frozen/thawed, sexed and unsexed semen samples from four Holstein bulls were randomly allocated to one of two different density gradient selection methods. Sperm quality (motility, concentration, morphology and membrane integrity) were evaluated and compared before and after sperm selection. Sperm motility and morphology improved (P < 0.005) after the sperm selection process with no differences between the two methods. For non-sexed semen, Percoll gradient increased the mean (± SEM) percentage of sperm recovered (57.3 ± 2.8) compared to Isolate (46.0 ± 1.8; P < 0.01). However, membrane integrity was higher after Isolate than Percoll (sexed semen: 41.0 ± 0.6 vs. 38.8 ± 0.8 and non-sexed semen 60.8 ± 1.6 vs. 58.8 ± 0.5; P < 0.05). The percentage of blastocysts produced was higher when either sexed or non-sexed semen was selected by Isolate (14.0 ± 1.0; 22.0 ± 1.1) than by Percoll (10.5 ± 1.5; 17.0 ± 2.1, respectively; P < 0.05). In summary, Isolate was a more effective method for the recovery of high quality sperm for in vitro fertilization embryo production.

  20. Evaluation of human sperm chromatin status after selection using a modified Diff-Quik stain indicates embryo quality and pregnancy outcomes following in vitro fertilization.

    PubMed

    Tavares, R S; Silva, A F; Lourenço, B; Almeida-Santos, T; Sousa, A P; Ramalho-Santos, J

    2013-11-01

    Sperm chromatin/DNA damage can be measured by a variety of assays. However, it has been reported that these tests may lose prognostic value in Assisted Reproductive Technology (ART) cycles when assessed