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Stable transfection of fatty acid translocase (CD36) in a rat heart muscle cell line (H9c2)  

Microsoft Academic Search

Fatty acid translocase (FAT\\/CD36) is a mem- brane protein putatively involved in the transmembrane transport of long-chain fatty acids. We tested the hypothesis that expression of this protein in H9c2, a rat heart cell line normally not expressing FAT, would increase cellular palmi- tate uptake. We were able to stably transfect H9c2 cells with FAT, yielding 15 cell lines showing

Frans A. Van Nieuwenhoven; Joost J. F. P. Luiken; Yvonne F. De Jong; Paul A. Grimaldi; Ger J. Van der Vusse; Jan F. C. Glatz


The H9C2 cell line and primary neonatal cardiomyocyte cells show similar hypertrophic responses in vitro  

Microsoft Academic Search

Cardiac hypertrophy is a major risk factor for heart failure and associated patient morbidity and mortality. Research investigating\\u000a the aberrant molecular processes that occur during cardiac hypertrophy uses primary cardiomyocytes from neonatal rat hearts\\u000a as the standard experimental in vitro system. In addition, some studies make use of the H9C2 rat cardiomyoblast cell line,\\u000a which has the advantage of being

Sarah J. Watkins; Gillian M. Borthwick; Helen M. Arthur



Translocation of HSP27 to Cytoskeleton by Repetitive Hypoxia-Reoxygenation in the Rat Myoblast Cell Line, H9c2  

Microsoft Academic Search

We investigated the possible changes in the distribution of HSP27 after a brief hypoxia-reoxygenation stress in the rat myoblast cell line, H9c2, as a model of ischemic preconditioning. Cells were exposed to 4 cycles of 5 min. of hypoxia and 5 min. of reoxygenation. In the normoxic condition, HSP27 was exclusively found in the cytosolic fraction. After the hypoxia-reoxygenation cycle,

Kenji Sakamoto; Tetsuro Urushidani; Taku Nagao



Phosphoinositide 3-kinase accelerates autophagic cell death during glucose deprivation in the rat cardiomyocyte-derived cell line H9c2  

Microsoft Academic Search

We investigated cell death during glucose deprivation in rat cardiomyocyte-derived H9c2 cells. Electron microscopic analysis revealed accumulation of autophagic vacuoles during glucose deprivation. The addition of 3-methyladenine or LY294002, which are known to inhibit autophagosome formation, reduced cell death while Z-VAD-FMK, a caspase inhibitor, slightly affected cell death. Thus, cell death during glucose deprivation is not type I programmed cell

Toshihiko Aki; Kazuhito Yamaguchi; Tatsuya Fujimiya; Yoichi Mizukami



Activation of Rac1 Increases c-Jun NH 2Terminal Kinase Activity and DNA Fragmentation in a Calcium-Dependent Manner in Rat Myoblast Cell Line H9c2  

Microsoft Academic Search

We examined the role of intracellular Ca2+ in c-Jun NH2-terminal kinase (JNK) activation and DNA fragmentation in the rat myoblast cell line H9c2 using small GTP-binding protein Rac1. A constitutively active mutant of Rac1 (V12-Rac1) increased JNK-responsive gene expression 6-fold, although this increase was attenuated by the intracellular Ca2+ chelator BAPTA-AM. V12-Rac1 also increased the number of DNA fragmentated cells.

Motohiro Nishida; Taku Nagao; Hitoshi Kurose



Leptin affects adenylate cyclase activity in H9c2 cardiac cell line: effects of short- and long-term exposure  

Microsoft Academic Search

Leptin has been hypothesized to be a pathophysiologic link between obesity and cardiovascular diseases. Because the adenylate cyclase (AC) system is a main effector of ?-adrenergic receptors and leptin has been shown to modulate AC activity in other cell lines, a leptin impact on cardiac AC activity was hypothesized. Therefore, acute and chronic effects of leptin on a rat cardiac

Gennaro Illiano; Silvio Naviglio; Mario Pagano; Annamaria Spina; Emilio Chiosi; Michelangela Barbieri; Giuseppe Paolisso



Lysophosphatidylcholine induces arachidonic acid release and calcium overload in cardiac myoblastic H9c2 cells  

Microsoft Academic Search

Lysophosphatidylcholine (lyso-PC) and arachido- nate are products of phosphatidylcholine hydrolysis by phospholipase A 2 . In this study, the modulation of arachido- nate release by exogenous lyso-PC in rat heart myoblastic H9c2 cells was examined. Incubation of H9c2 cells with lyso-PC resulted in an enhanced release of arachidonate in both a time- and dose-dependent fashion. Lyso-PC species containing palmitoyl (C

Leonard S. Golfman; Norman J. Haughey; Jason T. Wong; Jenny Y. Jiang; Douglas Lee; Jonathan D. Geiger; Patrick C. Choy


Mitochondrial Nitric Oxide Localization in H9c2 Cells Revealed by Confocal Microscopy  

Microsoft Academic Search

This study shows the presence of all three nitric oxide synthases (NOSs) and NOS activity in H9c2 cells cultured under non-stimulated conditions. By using the 4,5 diaminofluoresceindiacetate (DAF-2DA) fluorimetric nitric oxide (NO•) detection system we observed NO• production in H9c2 cells. As revealed by confocal microscopy, NO• fluorescence colocalizes in mitochondria labeled with Mito-Tracker Red CM-H2Xros. Upon stimulation with acetylcholine

B. Zanella; N. Calonghi; E. Pagnotta; L. Masotti; C. Guarnieri



Arsenite retards the cardiac differentiation of rat cardiac myoblast H9c2 cells.  


It is well known that exposure to inorganic arsenic through groundwater leads not only to cancer and cardiovascular disease, but also to detrimental effects on development. In this study, we investigated the effects of arsenite on the cardiac differentiation of rat myoblast H9c2 cells. The cardiac differentiation of H9c2 cells cultured in media containing 1% fetal bovine serum and all-trans retinoic acid was confirmed by enhanced expression of cardiac troponin T (cTnT), the appearance of multinucleated cells, and cell cycle arrest at G0/G1 phase. Exposure of H9c2 cells to inorganic arsenite (As(III)) during cardiac differentiation suppressed the appearance of the morphological and biological characteristics observed in the cardiac phenotype of H9c2 cells. In addition, As(III) inhibited PKC? phosphorylation, which is detected in early-stage differentiation. These results suggest that As(III) retards the cardiac differentiation of H9c2 cells, at least partly, via the inhibition of PKC? phosphorylation. PMID:23727579

Sumi, Daigo; Abe, Kazusa; Himeno, Seiichiro



The enhancement of phosphatidylcholine biosynthesis by angiotensin II in H9c2 cells  

Microsoft Academic Search

The effect of angiotensin II on the biosynthesis of phosphatidylcholine in rat heart myoblastic (H9c2) cells was investigated. Cells were incubated with [methyl-3H]choline, and the labelling of phosphatidylcholine at different time intervals was examined. When cells were pretreated with angiotensin II, a significant increase in the labelling of phosphatidylcholine was observed. Analysis of the labelled phosphatidylcholine precursors indicated that the

Khai Tran; Ricky Y. K. Man; Patrick C. Choy



Chronic-alcohol exposure alters IGF1 signaling in H9c2 cells via changes in PKC delta  

Microsoft Academic Search

Previously, we have demonstrated that chronic-alcohol exposure alters insulin-like growth factor 1 (IGF1) signaling in adult rat heart cells. This report examines the effects of alcohol in vitro on the expression of protein kinase C (PKC) alpha, delta, and epsilon using the embryonic heart cell line, H9c2, and how this may be linked to changes in IGF1 signal transduction. Western

Richard Ila; Michele Solem



Vital imaging of H9c2 myoblasts exposed to tert-butylhydroperoxide – characterization of morphological features of cell death  

Microsoft Academic Search

BACKGROUND: When exposed to oxidative conditions, cells suffer not only biochemical alterations, but also morphologic changes. Oxidative stress is a condition induced by some pro-oxidant compounds, such as by tert-butylhydroperoxide (tBHP) and can also be induced in vivo by ischemia\\/reperfusion conditions, which is very common in cardiac tissue. The cell line H9c2 has been used as an in vitro cellular

Vilma A Sardão; Paulo J Oliveira; Jon Holy; Catarina R Oliveira; Kendall B Wallace



Icariin Protects Rat Cardiac H9c2 Cells from Apoptosis by Inhibiting Endoplasmic Reticulum Stress  

PubMed Central

Endoplasmic reticulum stress (ERS) is one of the mechanisms of apoptotic cell death. Inhibiting the apoptosis induced by ERS may be a novel therapeutic target in cardiovascular diseases. Icariin, a flavonoid isolated from Epimedium brevicornum Maxim, has been demonstrated to have cardiovascular protective effects, but its effects on ERS are unknown. In the present study, we focused on icariin and investigated whether it might protect the cardiac cell from apoptosis via inhibition of ERS. In H9c2 rat cardiomyoblast cells, pretreatment of icariin significantly inhibited cell apoptosis by tunicamycin, an ERS inducer. Icariin also decreased generation of reactive oxygen species (ROS), loss of mitochondrial membrane potential and activation of caspase-3. Moreover, icariin inhibited upregulation of endoplasmic reticulum markers, GRP78, GRP94 and CHOP, elicited by tunicamycin. These results indicated that icariin could protect H9c2 cardiomyoblast cells from ERS-mitochondrial apoptosis in vitro, the mechanisms may be associated with its inhibiting of GRP78, GRP94 and CHOP and decreasing ROS generation directly. It may be a potential agent for treating cardiovascular disease.

Zhang, Qiufang; Li, Hongliang; Wang, Shanshan; Liu, Ming; Feng, Yibin; Wang, Xuanbin



Icariin protects rat cardiac H9c2 cells from apoptosis by inhibiting endoplasmic reticulum stress.  


Endoplasmic reticulum stress (ERS) is one of the mechanisms of apoptotic cell death. Inhibiting the apoptosis induced by ERS may be a novel therapeutic target in cardiovascular diseases. Icariin, a flavonoid isolated from Epimedium brevicornum Maxim, has been demonstrated to have cardiovascular protective effects, but its effects on ERS are unknown. In the present study, we focused on icariin and investigated whether it might protect the cardiac cell from apoptosis via inhibition of ERS. In H9c2 rat cardiomyoblast cells, pretreatment of icariin significantly inhibited cell apoptosis by tunicamycin, an ERS inducer. Icariin also decreased generation of reactive oxygen species (ROS), loss of mitochondrial membrane potential and activation of caspase-3. Moreover, icariin inhibited upregulation of endoplasmic reticulum markers, GRP78, GRP94 and CHOP, elicited by tunicamycin. These results indicated that icariin could protect H9c2 cardiomyoblast cells from ERS-mitochondrial apoptosis in vitro, the mechanisms may be associated with its inhibiting of GRP78, GRP94 and CHOP and decreasing ROS generation directly. It may be a potential agent for treating cardiovascular disease. PMID:23999590

Zhang, Qiufang; Li, Hongliang; Wang, Shanshan; Liu, Ming; Feng, Yibin; Wang, Xuanbin



Antiadrenergic effect of adenosine involves connexin 43 turn-over in H9c2 cells.  


Connexin 43 (Cx43) is the major protein of cardiac ventricular gap junctions and is crucial to cell-cell communication and cardiac function. Several authors report that adrenergic stimulation affects Cx43 expression via protein kinase A (PKA) and MAPK-regulated pathways. Adenosine has been shown to exert direct antiadrenergic effects on the heart, protecting it from toxic effects of overstimulation. The aim of our study was to understand the involvement of Cx43 in the anti-adrenergic effect of adenosine on cardiomyocytes. H9c2 cardiomyoblast cells were treated with isoproterenol alone or in association with adenosine. Isoproterenol and adenosine co-treated H9c2 cells showed an increased amount of Cx43 phosphorylated on Ser368. This effect was adenosine A1 receptor-dependent via the activation of the protein kinase C (PKC). Interestingly, the phosphorylation of Cx43 facilitated the degradation of Cx43 through the ubiquitin-proteasome system, as demonstrated by the immunoprecipitation of pCx43 with ubiquitin. On the basis of these results we can hypothesize that the activation of PKC after adenosine A1 receptor stimulation increases Cx43 phosphorylation that is necessary for its ubiquitination and then degradation via the proteasome system. These data better underline new mechanism at the basis of antiadrenergic effects of adenosine. PMID:23834776

Popolo, Ada; Morello, Silvana; Sorrentino, Rosalinda; Pinto, Aldo



Mitochondrial nitric oxide localization in H9c2 cells revealed by confocal microscopy.  


This study shows the presence of all three nitric oxide synthases (NOSs) and NOS activity in H9c2 cells cultured under non-stimulated conditions. By using the 4,5 diaminofluoresceindiacetate (DAF-2DA) fluorimetric nitric oxide (NO(*)) detection system we observed NO(*) production in H9c2 cells. As revealed by confocal microscopy, NO(*) fluorescence colocalizes in mitochondria labeled with Mito-Tracker Red CM-H(2)Xros. Upon stimulation with acetylcholine (Ach), which increased NOS activity by 75%, the colocalization coefficient C(green) value, calculated as Pearson's correlation, increased from 0.07 to 0.10, demonstrating an augmented presence of NO(*) in mitochondria. Conversely, the presence of NO(*) in mitochondria decreased following cells pretreatment with l-MonoMethylArginine (L-NMMA), a competitive inhibitor of NOS activity, as indicated by the reduction of the C(green) value to 0.02. This work confirms that the presence of NO(*) in mitochondria can be modulated in response to different fluxes of NO(*). PMID:11798175

Zanella, B; Calonghi, N; Pagnotta, E; Masotti, L; Guarnieri, C



Autophagy plays an important role in Sunitinib-mediated cell death in H9c2 cardiac muscle cells  

SciTech Connect

Sunitinib, which is a multitargeted tyrosine-kinase inhibitor, exhibits antiangiogenic and antitumor activity, and extends survival of patients with metastatic renal-cell carcinoma (mRCC) and gastrointestinal stromal tumors (GIST). This molecule has also been reported to be associated with cardiotoxicity at a high frequency, but the mechanism is still unknown. In the present study, we observed that Sunitinib showed high anti-proliferative effect on H9c2 cardiac muscle cells measured by PI staining and the MTT assay. But apoptotic markers (PARP cleavage, caspase 3 cleavage and chromatin condensation) were uniformly negative in H9c2 cells after Sunitinib treatment for 48 h, indicating that another cell death pathway may be involved in Sunitinib-induced cardiotoxicity. Here we found Sunitinib dramatically increased autophagic flux in H9c2 cells. Acidic vesicle fluorescence and high expression of LC3-II in H9c2 cells identified autophagy as a Sunitinib-induced process that might be associated with cytotoxicity. Furthermore, knocking down Beclin 1 by RNA-interference to block autophagy in H9c2 cells revealed that the death rate was decreased when treated with Sunitinib in comparison to control cells. These results confirmed that autophagy plays an important role in Sunitinib-mediated H9c2 cells cytotoxicity. Taken together, the data presented here strongly suggest that autophagy is associated with Sunitinib-induced cardiotoxicity, and that inhibition of autophagy constitutes a viable strategy for reducing Sunitinib-induced cardiomyocyte death thereby alleviating Sunitinib cardiotoxicity.

Zhao Yuqin; Xue Tao; Yang Xiaochun; Zhu Hong; Ding Xiaofei; Lou Liming [Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058 (China); Lu Wei [Department of Chemistry and Institute of Medicinal Chemistry, East China Normal University, Shanghai, 200062 (China); Yang Bo, E-mail: [Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058 (China); He Qiaojun, E-mail: [Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058 (China); Center for Drug Safety Evaluation and Research of Zhejiang University, Hangzhou, 310058 (China)



C-reactive protein induces p53-mediated cell cycle arrest in H9c2 cardiac myocytes  

Microsoft Academic Search

C-reactive protein (CRP) is one of the most important biomarker for cardiovascular diseases. Recent studies have shown that CRP affects cell survival, differentiation and apoptosis. However, the effect of CRP on the cell cycle has not been studied yet. We investigated the cell cycle alterations and cellular mechanisms induced by CRP in H9c2 cardiac myocytes. Flow cytometry analysis showed that

Ji-Won Choi; Kyung Hye Lee; Soo Hyuk Kim; Taewon Jin; Beom Seob Lee; Jaewon Oh; Ho-Yeon Won; Soo Young Kim; Seok-Min Kang; Ji Hyung Chung



Silibinin protects H9c2 cardiac cells from oxidative stress and inhibits phenylephrine-induced hypertrophy: potential mechanisms.  


Cardiac hypertrophy is the main response of the heart to various extrinsic and intrinsic stimuli, and it is characterized by specific molecular and phenotypic changes. Recent in vitro and in vivo studies indicate the involvement of reactive oxygen species in the hypertrophic response. In this study, silibinin, a plant flavonolignan extracted from milk thistle with potent antioxidant activity, was evaluated for its effects in (a) preventing hydrogen peroxide (H2O2)-induced cellular damage and (b) blocking the phenylephrine-induced hypertrophic response. Using the in vitro model of embryonic rat heart-derived H9c2 cells, we showed that silibinin has a rather safe profile as concentrations up to 200?M did not affect cell viability. Pretreatment of H9c2 cells with silibinin resulted in better protection of H9c2 cells under conditions of H2O2-induced cellular stress compared to untreated cells as indicated by cell viability and DNA fragmentation assays. Furthermore, silibinin attenuated the phenylephrine-induced hypertrophic response as evidenced by the measurement of cell surface, up-regulation of atrial natriuretic peptide and increase of cellular protein levels. Moreover, silibinin repressed the phenylephrine-induced phosphorylation of ERK1/2 kinases, while it appeared to inhibit the weakly activated by phenylephrine phosphorylation of Akt. Based on our results, silibinin may attenuate the phenylephrine-induced hypertrophic response of H9c2 cells via antioxidant mechanisms involving mainly the inhibition of the intracellular signaling pathways mediated by ERK1/2 MAPKs and Akt. PMID:22818713

Anestopoulos, Ioannis; Kavo, Anthula; Tentes, Ioannis; Kortsaris, Alexandros; Panayiotidis, Mihalis; Lazou, Antigone; Pappa, Aglaia



Insulin-Like Growth Factor I Modulates Induction of Apoptotic Signaling in H9C2 Cardiac Muscle Cells  

Microsoft Academic Search

Insulin-like growth factor I (IGF-I) is an important survival growth factor that has been shown to inhibit apoptosis, but the effects of IGF-I on apoptotic signaling remain largely unknown. To investigate IGF-I actions on apoptosis of H9C2 cardiac muscle cells, we have defined the effects of IGF-I on Bcl-2, Bax, caspase 3, DNA fragmentation, and cell survival. The abundance of




Angiopoietin-1 protects H9c2 cells from H 2O 2-induced apoptosis through AKT signaling  

Microsoft Academic Search

Loss of cardiomyocytes by apoptosis is proposed to cause ventricular remodeling and heart failure. Reactive oxygen species-induced apoptosis of cardiomyocytes has been reported to play an important role in many types of pathological processes of the heart. We investigated whether angiopoietin-1 (Ang1) has direct cytoprotective effects on cardiomyocytes against oxidative stress.Cultured H9c2 cells (cardiomyocytes) were treated with hydrogen peroxide (H2O2).

Zuoyan Wang; Ming Cui; Lijie Sun; Zhuqing Jia; Yun Bai; Kangtao Ma; Fengrong Chen; Chunyan Zhou



Role of quercetin and its in vivo metabolites in protecting H9c2 cells against oxidative stress  

Microsoft Academic Search

The aim of this study was to investigate the potential of quercetin and two of its “in vivo“ metabolites, 3?-O-methyl quercetin and 4?-O-methyl quercetin, to protect H9c2 cardiomyoblasts against H2O2-induced oxidative stress. As limited data are available regarding the potential uptake and cellular effects of quercetin and its metabolites in cardiac cells, we have evaluated the cellular association\\/uptake of the

C. Angeloni; J. P. E. Spencer; E. Leoncini; P. L. Biagi; S. Hrelia



Etomoxir mediates differential metabolic channeling of fatty acid and glycerol precursors into cardiolipin in H9c2 cells  

Microsoft Academic Search

We examined the effect of etomoxir treatment on de novo cardiolipin (CL) biosynthesis in H9c2 cardiac myoblast cells. Etomoxir treatment did not affect the activi- ties of the CL biosynthetic and remodeling enzymes but caused a reduction in (1- 14 C)palmitic acid or (1- 14 C)oleic acid incorporation into CL. The mechanism was a decrease in fatty acid flux through

Fred Y. Xu; William A. Taylor; Jeffrey A. Hurd; Grant M. Hatch



Oxidative Stress Induces DNA Fragmentation and Caspase Activation Via the c-Jun NH 2-terminal Kinase Pathway in H9c2 Cardiac Muscle Cells  

Microsoft Academic Search

The aim of this study was to test the hypothesis that oxidative stress induces apoptosis in the H9c2 cardiac muscle cell line, and that signaling via mitogen-activated protein kinase (MAPK) pathways is involved. Three forms of oxidative stress were utilized: the superoxide generator menadione; hydrogen peroxide; or simulated ischemia followed by reperfusion. Relatively low concentrations of menadione (10?m) or H2O2(250?m)

Neil A. Turner; Fen Xia; Gohar Azhar; Xiaomin Zhang; Lixin Liu; Jeanne Y Wei



Preparation and Characterization of Selenium Incorporated Guar Gum Nanoparticle and Its Interaction with H9c2 Cells  

PubMed Central

This study deals with the preparation and characterization of selenium incorporated guar gum nanoparticle (SGG), and its effect on H9c2 cardiomyoblast. Herein, nanoprecipitation techniques had been employed for the preparation of SGG nanoparticle. The prepared nanoparticle had been subjected to various types of analytical techniques like transmission electron microscopy (TEM), X-ray diffraction (XRD) and particle size analysis to confirm the characteristics of nanoparticle as well as for selenium incorporation. Physical characterization of nanoparticle showed that the size of nanoparticles increase upto ?69–173 nm upon selenium incorporation from ?41–132 nm. Then the prepared nanoparticles were evaluated for its effect on H9c2 cells. In this regard, the effect of nanoparticle on various vital parameters of H9c2 cells was studied. Parameters like cell viability, uptake of selenium incorporated guar gum nanoparticle by the cells, effect of SGG on DNA integrity, apoptosis, reactive oxygen species generation, alteration in transmembrane potential of mitochondria and cytoskeletal integrity had been investigated. Viability results showed that up to 25 nM of SGG was safe (10.31%) but beyond that it induces cytotoxicity. Cellular uptake of selenium showed that cell permeability for SGG is significantly high compared to normal selenium (7.2 nM of selenium for 25 nM SGG compared with 5.2 nM selenium for 25 nM sodium selenite). There was no apoptosis with SGG and also it protects DNA from hydroxyl radical induced breakage. Likewise no adverse effect on mitochondria and cytoskeleton was observed for 25 nM of SGG. Overall results reveal that SGG is highly suitable for biomedical research application.

Soumya, Rema Sreenivasan; Vineetha, Vadavanath Prabhakaran; Reshma, Premachandran Latha; Raghu, Kozhiparambil Gopalan



Preparation and characterization of selenium incorporated guar gum nanoparticle and its interaction with h9c2 cells.  


This study deals with the preparation and characterization of selenium incorporated guar gum nanoparticle (SGG), and its effect on H9c2 cardiomyoblast. Herein, nanoprecipitation techniques had been employed for the preparation of SGG nanoparticle. The prepared nanoparticle had been subjected to various types of analytical techniques like transmission electron microscopy (TEM), X-ray diffraction (XRD) and particle size analysis to confirm the characteristics of nanoparticle as well as for selenium incorporation. Physical characterization of nanoparticle showed that the size of nanoparticles increase upto ?69-173 nm upon selenium incorporation from ?41-132 nm. Then the prepared nanoparticles were evaluated for its effect on H9c2 cells. In this regard, the effect of nanoparticle on various vital parameters of H9c2 cells was studied. Parameters like cell viability, uptake of selenium incorporated guar gum nanoparticle by the cells, effect of SGG on DNA integrity, apoptosis, reactive oxygen species generation, alteration in transmembrane potential of mitochondria and cytoskeletal integrity had been investigated. Viability results showed that up to 25 nM of SGG was safe (10.31%) but beyond that it induces cytotoxicity. Cellular uptake of selenium showed that cell permeability for SGG is significantly high compared to normal selenium (7.2 nM of selenium for 25 nM SGG compared with 5.2 nM selenium for 25 nM sodium selenite). There was no apoptosis with SGG and also it protects DNA from hydroxyl radical induced breakage. Likewise no adverse effect on mitochondria and cytoskeleton was observed for 25 nM of SGG. Overall results reveal that SGG is highly suitable for biomedical research application. PMID:24098647

Soumya, Rema Sreenivasan; Vineetha, Vadavanath Prabhakaran; Reshma, Premachandran Latha; Raghu, Kozhiparambil Gopalan



[Recombinant adenovirus overexpressing nkx2.5 protects H9c2 cells against H2O2-induced apoptosis].  


To study the function and potential application of nkx2.5, a critical gene for heart development, we constructed a recombinant adenovirus overexpressing nkx2.5 gene (Ad-Nkx2.5) with the AdEasy system. To evaluate the effect and mechanism of Ad-Nkx2.5 against oxidative injury, the H9c2 myocardial cells were infected with the recombinant adenoviruses Ad-Nkx2.5 or Ad-EGFP, and subsequently exposed to H2O2 to induce apoptosis. The anti-apoptotic potential of Ad-Nkx2.5 was validated by MTT assay for cell viability, Hoechst33342 staining for cellular morphology, and immunoblotting for caspase-3 activity. Ad-Nkx2.5 infection led to an increased survival rate of H9c2 cells and decreased the amount of caspase-3 in an active form. Additionally, overexpression of Nkx2.5 inhibited the release of cytochrome C from the mitochondria into the cytosol. Mechanismic studies showed that Nkx2.5 upregulated bcl-2 gene expression and significantly repressed H2O2-induced expression of bax detected by Real-time PCR. Additionally, H2O2 treatment did not affect the nuclear localization of Nkx2.5. These findings indicate that adenovirus-mediated nkx2.5 gene transfer exerted a protective effect on H9c2 cells against H2O2-induced apoptosis via mitochondrial pathway, and the Nkx2.5-mediated expression modulation of apoptosis-associated genes could be involved in this event. PMID:23311140

Li, Tao; Jiang, Kesheng; Ruan, Qin; Liu, Zhiqiang



TanshinoneIIA and Cryptotanshinone Protect against Hypoxia-Induced Mitochondrial Apoptosis in H9c2 Cells  

PubMed Central

Mitochondrial apoptosis pathway is an important target of cardioprotective signalling. Tanshinones, a group of major bioactive compounds isolated from Salvia miltiorrhiza, have been reported with actions against inflammation, oxidative stress, and myocardial ischemia reperfusion injury. However, the actions of these compounds on the chronic hypoxia-related mitochondrial apoptosis pathway have not been investigated. In this study, we examined the effects and molecular mechanisms of two major tanshonones, tanshinone IIA (TIIA) and cryptotanshinone (CT) on hypoxia induced apoptosis in H9c2 cells. Cultured H9c2 cells were treated with TIIA and CT (0.3 and 3 ??) 2 hr before and during an 8 hr hypoxic period. Chronic hypoxia caused a significant increase in hypoxia inducible factor 1? expression and the cell late apoptosis rate, which was accompanied with an increase in caspase 3 activity, cytochrome c release, mitochondria membrane potential and expression of pro-apoptosis proteins (Bax and Bak). TIIA and CT (0.3 and 3 ??), in concentrations without affecting the cell viability, significantly inhibited the late apoptosis and the changes of caspase 3 activity, cytochrome c release, and mitochondria membrane potential induced by chronic hypoxia. These compounds also suppressed the overexpression of Bax and reduced the ratio of Bax/Bcl-2. The results indicate that TIIA and CT protect against chronic hypoxia induced cell apoptosis by regulating the mitochondrial apoptosis signaling pathway, involving inhibitions of mitochondria hyperpolarization, cytochrome c release and caspase 3 activity, and balancing anti- and pro-apoptotic proteins in Bcl-2 family proteins.

Jin, Hyou-Ju; Xie, Xiao-Liang; Ye, Ji-Ming; Li, Chun-Guang



Natural (ghrelin) and synthetic (hexarelin) GH secretagogues stimulate H9c2 cardiomyocyte cell proliferation  

Microsoft Academic Search

Recent experimental data demonstrate cardiovascular effects of the GH secretagogues (GHSs) hexarelin and ghrelin, the proposed natural ligand for the GHS receptor. Moreover, specific cardiac binding sites for GHSs have been suggested. The aim of the present study was to investigate if the natural ligand ghrelin and synthetic GHS peptide hexarelin and analogues have direct effects on the cardiomyocyte cell

I Pettersson; G Muccioli; R Granata; R Deghenghi; E Ghigo; C Ohlsson; J Isgaard



Sodium nitroprusside induces apoptosis of H9C2 cardiac muscle cells in a c-Jun N-terminal kinase-dependent manner  

Microsoft Academic Search

Sodium nitroprusside (SNP) induces apoptosis in H9C2 cardiac muscle cells. Treatment with an exogenous NO donor SNP (2 mM) to H9C2 cells resulted in apoptotic morphological changes; a bright blue-fluorescent condensed nuclei and chromatin fragmentation by fluorescence microscope of Hoechst 33258-staining. The activity of caspase-3 like protease was increased during SNP-induced cell death. However, the activity of caspase-1 like protease

Han-jung Chae; Hong-seob So; Soo-wan Chae; Ji-sun Park; Myung-sun Kim; Jay-min Oh; Yeun-tai Chung; Sei-hoon Yang; Eun-taik Jeong; Hyung-min Kim; Rae-kil Park; Hyung-Ryong Kim



Quercetin protects the hydrogen peroxide-induced apoptosis via inhibition of mitochondrial dysfuntion in H9c2 cardiomyoblast cells  

Microsoft Academic Search

Quercetin possesses a broad range of pharmacological properties, including protection of LDL from oxidation. However, little is known about the mechanism by which quercetin rescues cardiomyoblasts from oxidative damage. This study was designed to investigate the protective mechanism of quercetin on H2O2-induced toxicity of H9c2 cardiomyoblasts. Oxidative stress, such as H2O2, ZnCl2, and menadione, significantly decreased the viability of H9c2

Channy Park; Hong-Seob So; Chang-Ho Shin; Seung-Hwa Baek; Byung-Soon Moon; Sun-Ho Shin; Ho-Seob Lee; Dong-Wook Lee



Role of NADPH oxidase in H9c2 cardiac muscle cells exposed to simulated ischaemia-reperfusion.  


Oxidative stress is associated with several cardiovascular pathologies, including hypertension, cardiac hypertrophy and heart failure. Although oxidative stress is also increased after ischaemia-reperfusion (I/R), little is known about the role and the activation mechanisms, in cardiac myocytes under these conditions, of NADPH oxidase, a superoxide-producing enzyme. We found that rat cardiac muscle cells (H9c2) subjected to an in vitro simulated ischaemia (substrate-free medium plus hypoxia) followed by 'reperfusion', displayed increased reactive oxygen species (ROS) production attributable to a parallel increase of NADPH oxidase activity. Our investigation on mechanisms responsible for NADPH oxidase activation showed a contribution of both the increase of NOX2 expression and p47(phox) translocation to the membrane. We also found that the increase of NADPH oxidase activity was associated with higher levels of lipid peroxidation, the activation of redox-sensitive kinases, in particular ERK and JNK, and with cell death. Diphenyleneiodonium (DPI), a flavoprotein inhibitor used as NADPH oxidase inhibitor, prevented I/R-induced ROS formation in treated cells, together with the related lipoperoxidative damage, and JNK phosphorylation without affecting ERK activation, resulting in protection against cell death. Our results provide evidence that NADPH oxidase is a key enzyme involved in I/R-induced oxidant generation and suggest it can be a possible target in cardioprotective strategies against I/R injury, a condition of great importance in human pathology. PMID:18754815

Borchi, Elisabetta; Parri, Matteo; Papucci, Laura; Becatti, Matteo; Nassi, Niccolò; Nassi, Paolo; Nediani, Chiara



Etomoxir mediates differential metabolic channeling of fatty acid and glycerol precursors into cardiolipin in H9c2 cells.  


We examined the effect of etomoxir treatment on de novo cardiolipin (CL) biosynthesis in H9c2 cardiac myoblast cells. Etomoxir treatment did not affect the activities of the CL biosynthetic and remodeling enzymes but caused a reduction in [1-14C]palmitic acid or [1-14C]oleic acid incorporation into CL. The mechanism was a decrease in fatty acid flux through the de novo pathway of CL biosynthesis via a redirection of lipid synthesis toward 1,2-diacyl-sn-glycerol utilizing reactions mediated by a 35% increase (P < 0.05) in membrane phosphatidate phosphohydrolase activity. In contrast, etomoxir treatment increased [1,3-3H]glycerol incorporation into CL. The mechanism was a 33% increase (P < 0.05) in glycerol kinase activity, which produced an increased glycerol flux through the de novo pathway of CL biosynthesis. Etomoxir treatment inhibited 1,2-diacyl-sn-glycerol acyltransferase activity by 81% (P < 0.05), thereby channeling both glycerol and fatty acid away from 1,2,3-triacyl-sn-glycerol utilization toward phosphatidylcholine and phosphatidylethanolamine biosynthesis. In contrast, etomoxir inhibited myo-[3H]inositol incorporation into phosphatidylinositol and the mechanism was an inhibition in inositol uptake. Etomoxir did not affect [3H]serine uptake but resulted in an increased formation of phosphatidylethanolamine derived from phosphatidylserine. The results indicate that etomoxir treatment has diverse effects on de novo glycerolipid biosynthesis from various metabolic precursors. In addition, etomoxir mediates a distinct and differential metabolic channeling of glycerol and fatty acid precursors into CL. PMID:12576524

Xu, Fred Y; Taylor, William A; Hurd, Jeffrey A; Hatch, Grant M



N-acetylcysteine prevents glucose\\/glucose oxidase-induced oxidative stress, mitochondrial damage and apoptosis in H9c2 cells  

Microsoft Academic Search

AimsHigh blood glucose may auto-oxidize and generate free radicals, which are proposed to induce apoptosis in cardiac cells. The aim of the present study was to investigate the cell damage induced by glucose\\/glucose oxidase-dependent oxidative stress and the protective effect of N-acetylcysteine (NAC) on H9c2 cardiac muscle cells.

Santosh Kumar; Sandhya L. Sitasawad



Doxorubicin-induced cardiotoxicity: direct correlation of cardiac fibroblast and H9c2 cell survival and aconitase activity with heat shock protein 27.  


The use of doxorubicin (Dox) and its derivatives as chemotherapeutic drugs to treat patients with cancer causes dilated cardiomyopathy and congestive heart failure due to Dox-induced cardiotoxicity. In this work, using heat shock factor-1 wild-type (HSF-1(+/+)) and HSF-1 knockout (HSF-1(-/-)) mouse fibroblasts and embryonic rat heart-derived cardiac H9c2 cells, we show that the magnitude of protection from Dox-induced toxicity directly correlates with the level of the heat shock protein 27 (HSP27). Western blot analysis of normal and heat-shocked cells showed the maximum expression of HSP27 in heat-shocked cardiac H9c2 cells and no HSP27 in HSF-1(-/-) cells (normal or heat-shocked). Correspondingly, the cell viability, measured [with (3,4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay] after treatment with various concentrations of Dox, was the highest in heat-shocked H9c2 cells and the lowest in HSF-1(-/-) cells. Depleting HSP27 in cardiac H9c2 cells by small interfering (si)RNA also reduced the viability against Dox, confirming that HSP27 does protect cardiac cells against the Dox-induced toxicity. The cells that have lower HSP27 levels such as HSF-1(-/-), were found to be more susceptible for aconitase inactivation. Based on these results we propose a novel mechanism that HSP27 plays an important role in protecting aconitase from Dox-generated O(2)*(-), by increasing SOD activity. Such a protection of aconitase by HSP27 eliminates the catalytic recycling of aconitase released Fe(II) and its deleterious effects in cardiac cells. PMID:17873025

Turakhia, Samir; Venkatakrishnan, C D; Dunsmore, Kathy; Wong, Hector; Kuppusamy, Periannan; Zweier, Jay L; Ilangovan, Govindasamy



Hyaluronic acid-dependent protection in H9C2 cardiomyocytes: a cell model of heart ischemia-reperfusion injury and treatment.  


Hyaluronic acid (HA), a glycosaminoglycan with high molecular weight, has been reported to promote cell proliferation and serves as an important extracellular matrix component. The aim of this study was to in vitro investigate whether HA is able to reduce reactive oxygen species (ROS)-induced heart ischemia-reperfusion injury and activate the cardiomyocyte's damage surveillance systems. Accordingly, rattus cardiomyocyte line, H9C2, was treated with H(2)O(2) as a heart ischemia-reperfusion model followed by incubation with low molecular weight hyaluronan (LMW-HA, 100 kDa) or high molecular weight hyaluronan (HMW-HA, 1000 kDa) and proteomic analysis was performed to investigate the physiologic protection of HA in H(2)O(2)-induced ischemia-reperfusion in cardiomyocyte. Our data demonstrated that HA treatment does protect cardiomyocyte in the ROS-induced ischemia-reperfusion model and the molecular weight of HA is a crucial factor. HMW-HA has been shown to significantly facilitate cell migration and wound healing via cytoskeletal rearrangement. Additionally, 2D-DIGE combined MALDI-TOF/TOF analysis showed that HMW-HA might modulate biosynthetic pathways, cell migration, cell outgrowth and protein folding to stimulate wound healing as well as prevent these ischemia-reperfusion-damaged cardiomyocytes from cell death. To our knowledge, we report for the first time the cell repair mechanism of HMW-HA against ischemia-reperfusion-damage in cardiomyocytes based on cell biology and proteomic analysis. PMID:23178681

Law, Ching-Hsuan; Li, Ji-Min; Chou, Hsiu-Chuan; Chen, Yu-Hua; Chan, Hong-Lin



[The up-regulation of hypoxia inducible factor-1? by hypoxic postconditioning reduces hypoxia/reoxygenation-induced injury in heart-derived H9c2 cells].  


The aim of the present study was to explore the effects of hypoxic postconditioning (PostC) on heart-derived H9c2 cells injury induced by hypoxia/reoxygenation (H/R) and the expression of hypoxia inducible factor-1? (HIF-1?), and to analyze the relationship between them. Cultured H9c2 cardiac muscle cells were subjected to 3-hour hypoxia and 2-hour reoxygenation to simulate ischemia and reperfusion, or underwent 3 cycles of 5-min reoxygenation and 5-min hypoxia preceding the long reoxygenation to simulate ischemic postconditioning. Cell viability, lactate dehydrogenase (LDH) activity, and caspase-3 activity were detected respectively to investigate the cell injury induced by H/R. The level of HIF-1? mRNA was measured by real-time PCR. Western blot was used to determine HIF-1? protein level. The results showed that postconditioning significantly increased H9c2 cell viability, reduced the activity of LDH and caspase-3. Simultaneously, postconditioning up-regulated the HIF-1? protein level. Moreover, after DMOG, an inhibitor of proline hydroxylase (PHD) which targeted to HIF-1? degradation, was used to stabilize HIF-1? protein level, the reduction of H9c2 cells injury was comparable to that by postconditioning. There was a significant linear positive relationship between HIF-1? protein level and cell viability (r = 0.743, P < 0.01). After HIF-1? gene was silenced by siRNA, the cardio-protective effects of postconditioning was significantly weakened. These data suggest that up-regulation of HIF-1? plays an important role in the cardio-protection of postconditioning. PMID:23788186

Zhao, Huan-Xin; Li, Xiao-Yu; Yao, Hong; Wu, Ye; Yang, Rong; Zhao, Rong-Rui; Liu, Hui-Rong



Protective effects of steroids from Allium chinense against H2O2-induced oxidative stress in rat cardiac H9C2 cells.  


Allium chinense, a traditional herbal medicine, has been used for the treatment of cardiovascular diseases for hundreds of years. In this study, A. chinense steroids (ACSs) including three steroidal glycosides and their parent aglycones were isolated from the bulbs of A. chinense. For the first time, their cardioprotective effects were evaluated in cultured rat cardiac H9C2 cells by pretreatment with ACSs for 24 h before exposure to 0.2 mm H(2)O(2). The results showed the cell viability decreased markedly when H9C2 cells were incubated with 0.2 mm H(2)O(2) alone for 2 h, while the cell lipid peroxidation (estimated by the excessive production of nitric oxide and malondialdehyde) and intracellular free calcium concentration ([Ca(2+)](i)) increased significantly. The addition of 20 microm (below the toxic concentration) of ACSs notably attenuated the cellular injury induced by H(2)O(2). The effects of ACSs in our experiments were similar to those of nimodipine, a clinically applied calcium channel blocker. Preliminary analysis of the structure-activity relationship indicated that ACSs with a spirostane-type skeleton exhibited stronger protection than that with a furostane-type skeleton, and glycosylation of the steroids could substantially lower the protective activities. The above results suggested the protective effects of steroids originated from A. chinense on the oxidative injury of H9C2 cells and ACSs may have potential for preventing cardiac injuries induced by oxidative stress. PMID:19653197

Ren, Gang; Qiao, Hong Xiang; Yang, Jun; Zhou, Chang Xin



SIRT2 is a negative regulator of anoxia–reoxygenation tolerance via regulation of 14-3-3 ? and BAD in H9c2 cells  

Microsoft Academic Search

Knockdown or inhibition of SIRT2 enhances biological stress-tolerance. We extend this phenotype showing that SIRT2 knockdown reduces anoxia–reoxygenation injury in H9c2 cells. Gene array analysis following SIRT2 siRNA knockdown identifies 14-3-3 ? as the most robustly induced gene. SIRT2 knockdown evokes induction of this chaperone, facilitating cytosolic sequestration of BAD with a corresponding reduction in mitochondrial BAD localization. Concurrent siRNA

Edward G. Lynn; Christopher J. McLeod; Jeffrey P. Gordon; Jianjun Bao; Michael N. Sack



ARC Inhibits Cytochrome c Release From Mitochondria and Protects Against Hypoxia-Induced Apoptosis in Heart-Derived H9c2 Cells  

Microsoft Academic Search

Ischemia induces apoptosis as well as necrosis of cardiac myocytes. We recently reported the cloning of a cDNA that encodes an apoptotic inhibitor, ARC, that is expressed predominantly in cardiac and skeletal muscle. In the present study, we examined the ability of ARC to protect rat embryonic heart- derived H9c2 cells from apoptosis induced by hypoxia, a component of ischemia.

Daryoush Ekhterae; Zhiwu Lin; Martha S. Lundberg; Michael T. Crow; Frank C. Brosius; Gabriel Nunez



Activation of the p38 MAPK/NF-?B pathway contributes to doxorubicin-induced inflammation and cytotoxicity in H9c2 cardiac cells.  


A number of studies have demonstrated that inflammation plays a role in doxorubicin (DOX)-induced cardiotoxicity. However, the molecular mechanism by which DOX induces cardiac inflammation has yet to be fully elucidated. The present study aimed to investigate the role of the p38 mitogen-activated protein kinase (MAPK)/nuclear factor-?B (NF-?B) pathway in DOX-induced inflammation and cytotoxicity. The results of our study demonstrated that the exposure of H9c2 cardiac cells to DOX reduced cell viability and stimulated an inflammatory response, as demonstrated by an increase in the levels of interleukin-1? (IL-1?) and IL-6, as well as tumor necrosis factor-? (TNF-?) production. Notably, DOX exposure induced the overexpression of phosphorylated p38 MAPK and phosphorylation of the NF-?B p65 subunit, which was markedly inhibited by SB203580, a specific inhibitor of p38 MAPK. The inhibition of NF-?B by pyrrolidine dithiocarbamate (PDTC), a selective inhibitor of NF-?B, significantly ameliorated DOX-induced inflammation, leading to a decrease in the levels of IL-1? and IL-6, as well as TNF-? production in H9c2 cells. The pretreatment of H9c2 cells with either SB203580 or PDTC before exposure to DOX significantly attenuated DOX-induced cytotoxicity. In conclusion, our study provides novel data demonstrating that the p38 MAPK/NF-?B pathway is important in the induction of DOX-induced inflammation and cytotoxicity in H9c2 cardiac myocytes. PMID:23807148

Guo, Run-Min; Xu, Wen-Ming; Lin, Jian-Cong; Mo, Li-Qiu; Hua, Xiao-Xiao; Chen, Pei-Xi; Wu, Keng; Zheng, Dong-Dan; Feng, Jian-Qiang



NADPH-oxidase-dependent Superoxide Production by Myocyte-derived H9c2 Cells: Influence of Ischemia, Heat Shock, Cycloheximide and Cytochalasin D  

Microsoft Academic Search

Extracellular oxygen radicals produced by H9c2 rat heart cells in monolayer cultures during ischemia and subsequent reoxygenation were monitored using the luminol-horseradish peroxidase-enhanced chemiluminescence technique. As expected, the photon count diminishes during ischemia but again rapidly attains normal values following reoxygenation. In the presence of superoxide dismutase, this photon emission is repressed, as is also the case in the presence

Jan E. M Souren; Cor Van Der Mast; Roeland Van Wijk



Propofol protects against hydrogen peroxide-induced injury in cardiac H9c2 cells via Akt activation and Bcl2 up-regulation  

Microsoft Academic Search

Propofol is a widely used intravenous anesthetic agent with antioxidant properties secondary to its phenol based chemical structure. Treatment with propofol has been found to attenuate oxidative stress and prevent ischemia\\/reperfusion injury in rat heart. Here, we report that propofol protects cardiac H9c2 cells from hydrogen peroxide (H2O2)-induced injury by triggering the activation of Akt and a parallel up-regulation of

Baohua Wang; Jayant Shravah; Honglin Luo; Koen Raedschelders; David D. Y. Chen; David M. Ansley



Resveratrol protects ROS-induced cell death by activating AMPK in H9c2 cardiac muscle cells  

Microsoft Academic Search

Resveratrol, one of polyphenols derived from red wine, has been shown to protect against cell death, possibly through the\\u000a association with several signaling pathways. Currently numerous studies indicate that cardiovascular diseases are linked to\\u000a the release of intracellular reactive oxygen species (ROS) often generated in states such as ischemia\\/reperfusion injury.\\u000a In the present study, we investigated whether resveratrol has the

Jin-Taek Hwang; Dae Young Kwon; Ock Jin Park; Myung Sunny Kim



Testosterone protects female embryonic heart H9c2 cells against severe metabolic stress by activating estrogen receptors and up-regulating IES SUR2B.  


A recent clinical study demonstrated that a testosterone supplementation improves functional capacity in elderly female patients suffering from heart failure. These findings prompted us to consider possible mechanisms of testosterone-induced cardioprotection in females. To address this question we have used a pure female population of rat heart embryonic H9c2 cells. Pre-treatment of cells with testosterone for 24h significantly increased survival of H9c2 cells exposed to 2,4-dinitrophenol (DNP), an inhibitor of oxidative phosphorylation. These cells expressed low level of androgen receptors and the effect of testosterone was not modified by hydroxyflutamide, an antagonist of androgen receptor. In contrast, cyclohexamide, an inhibitor of protein biosynthesis, and tamoxifene, a partial agonist of estrogen receptors, abolished cardioprotection afforded by testosterone. In addition, finasteride, an inhibitor of 5?-reductase, and anastrazole, an inhibitor of ?-aromatase, also blocked testosterone-induced cytoprotection. Real time RT-PCR revealed that testosterone did not regulate the expression of nine subunits and accessory proteins of sarcolemmal ATP-sensitive K(+) (K(ATP)) channels. On the other hand, testosterone, as well as 17?-estradiol, up-regulated a putative mitochondrial K(ATP) channel subunit, mitochondrial sulfonylurea receptor 2B intraexonics splice variant (IES SUR2B), without affecting expression of IES SUR2A. Tamoxifene inhibited testosterone-induced up-regulation of IES SUR2B without affecting IES SUR2A. In conclusion, this study has shown that testosterone protect female embryonic heart H9c2 cells against severe metabolic stress by its conversion into metabolites that activate estrogen receptors and up-regulate IES SUR2B. PMID:23085378

Ballantyne, Thomas; Du, Qingyou; Jovanovi?, Sofija; Neemo, Andrew; Holmes, Robert; Sinha, Sharabh; Jovanovi?, Aleksandar



NADPH oxidase/ROS-dependent PYK2 activation is involved in TNF-?-induced matrix metalloproteinase-9 expression in rat heart-derived H9c2 cells.  


TNF-? plays a mediator role in the pathogenesis of chronic heart failure contributing to cardiac remodeling and peripheral vascular disturbances. The implication of TNF-? in inflammatory responses has been shown to be mediated through up-regulation of matrix metalloproteinase-9 (MMP-9). However, the detailed mechanisms of TNF-?-induced MMP-9 expression in rat embryonic-heart derived H9c2 cells are largely not defined. We demonstrated that in H9c2 cells, TNF-? induced MMP-9 mRNA and protein expression associated with an increase in the secretion of pro-MMP-9. TNF-?-mediated responses were attenuated by pretreatment with the inhibitor of ROS (N-acetyl-l-cysteine, NAC), NADPH oxidase [apocynin (APO) or diphenyleneiodonium chloride (DPI)], MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), NF-?B (Bay11-7082), or PYK2 (PF-431396) and transfection with siRNA of TNFR1, p47(phox), p42, p38, JNK1, p65, or PYK2. Moreover, TNF-? markedly induced NADPH oxidase-derived ROS generation in these cells. TNF-?-enhanced p42/p44 MAPK, p38 MAPK, JNK1/2, and NF-?B (p65) phosphorylation and in vivo binding of p65 to the MMP-9 promoter were inhibited by U0126, SB202190, SP600125, NAC, DPI, or APO. In addition, TNF-?-mediated PYK2 phosphorylation was inhibited by NAC, DPI, or APO. PYK2 inhibition could reduce TNF-?-stimulated MAPKs and NF-?B activation. Thus, in H9c2 cells, we are the first to show that TNF-?-induced MMP-9 expression is mediated through a TNFR1/NADPH oxidase/ROS/PYK2/MAPKs/NF-?B cascade. We demonstrated that NADPH oxidase-derived ROS generation is involved in TNF-?-induced PYK2 activation in these cells. Understanding the regulation of MMP-9 expression and NADPH oxidase activation by TNF-? on H9c2 cells may provide potential therapeutic targets of chronic heart failure. PMID:23774252

Yang, Chuen-Mao; Lee, I-Ta; Hsu, Ru-Chun; Chi, Pei-Ling; Hsiao, Li-Der



ZAK re-programs atrial natriuretic factor expression and induces hypertrophic growth in H9c2 cardiomyoblast cells  

Microsoft Academic Search

Various intracellular or intercellular stimuli have been associated with the development of cardiac cell hypertrophy. However, the mechanisms underlying this association are not completely understood. In a previous study we determined that ZAK mRNA expression is abundant in heart. ZAK is a mitogen-activated protein kinase kinase kinase (MAP3K) that activates the stress-activated protein kinase\\/c-jun N-terminal kinase pathway and activates NF-?B.

Chih-Yang Huang; Pin Ju Chueh; Chien-Tang Tseng; Kuan-Yu Liu; Hsiao-Ya Tsai; Wei-Wen Kuo; Ming-Yung Chou; Jaw-Ji Yang



Ghrelin protects against cobalt chloride-induced hypoxic injury in cardiac H9c2 cells by inhibiting oxidative stress and inducing autophagy.  


Ghrelin is a multifunctional peptide that actively protects against cardiovascular ischemic diseases, but the underlying mechanisms are unclear. We used CoCl(2) to mimic hypoxic conditions in cardiac H9c2 cells in order to study the mechanism by which ghrelin protects cardiac myocytes against hypoxic injury by regulating the content of intracellular ROS and autophagy levels. Cell apoptosis and necrosis were evaluated by the flow cytometry assay, Hoechst staining, and LDH activity. Cell viability was detected by the WST-1 assay; ROS levels were assessed using DCFH2-DA; and Nox1, catalase and Mn-SOD were assayed by real-time PCR and activity assays. LC3II was measured by Western blot analysis. We observed that CoCl(2) induced apoptosis and death of H9c2 cells in a dose- and time-dependent manner. This was characterized by an increase in cell apoptosis, LDH activity, ROS content, Nox1 expression, and autophagy levels and a decrease in cell viability, catalase, and Mn-SOD activities. Ghrelin treatment significantly attenuated CoCl(2)-induced hypoxic injury by decreasing cell apoptosis, LDH activity, ROS content, and Nox1 expression and increasing cell viability, autophagy levels, catalase, and Mn-SOD mRNA levels and activities. Further experiments revealed that inhibiting autophagy using 3-MA or AMPK pathway with compound C almost abrogated the induction of ghrelin in autophagy. This was associated with a decrease in cell viability and an increase in LDH activity. Our results indicate that ghrelin protected cardiac myocytes against CoCl(2)-induced hypoxic injury by decreasing Nox1 expression, increasing the expression and activity of endogenous antioxidant enzymes, and inducing protective autophagy in an AMPK-dependent manner. PMID:23000094

Tong, Xin-Xin; Wu, Dan; Wang, Xue; Chen, Hua-Li; Chen, Jia-Xiang; Wang, Xiao-Xiao; Wang, Xu-Lei; Gan, Lu; Guo, Zhi-Yun; Shi, Gui-Xiu; Zhang, Yi-Zheng; Jiang, Wei



Pan-histone deacetylase inhibitors regulate signaling pathways involved in proliferative and pro-inflammatory mechanisms in H9c2 cells  

PubMed Central

Background We have shown previously that pan-HDAC inhibitors (HDACIs) m-carboxycinnamic acid bis-hydroxamide (CBHA) and trichostatin A (TSA) attenuated cardiac hypertrophy in BALB/c mice by inducing hyper-acetylation of cardiac chromatin that was accompanied by suppression of pro-inflammatory gene networks. However, it was not feasible to determine the precise contribution of the myocytes- and non-myocytes to HDACI-induced gene expression in the intact heart. Therefore, the current study was undertaken with a primary goal of elucidating temporal changes in the transcriptomes of cardiac myocytes exposed to CBHA and TSA. Results We incubated H9c2 cardiac myocytes in growth medium containing either of the two HDACIs for 6h and 24h and analyzed changes in gene expression using Illumina microarrays. H9c2 cells exposed to TSA for 6h and 24h led to differential expression of 468 and 231 genes, respectively. In contrast, cardiac myocytes incubated with CBHA for 6h and 24h elicited differential expression of 768 and 999 genes, respectively. We analyzed CBHA- and TSA-induced differentially expressed genes by Ingenuity Pathway (IPA), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Core_TF programs and discovered that CBHA and TSA impinged on several common gene networks. Thus, both HDACIs induced a repertoire of signaling kinases (PTEN-PI3K-AKT and MAPK) and transcription factors (Myc, p53, NFkB and HNF4A) representing canonical TGF?, TNF-?, IFN? and IL-6 specific networks. An overrepresentation of E2F, AP2, EGR1 and SP1 specific motifs was also found in the promoters of the differentially expressed genes. Apparently, TSA elicited predominantly TGF?- and TNF-?-intensive gene networks regardless of the duration of treatment. In contrast, CBHA elicited TNF-? and IFN? specific networks at 6 h, followed by elicitation of IL-6 and IFN?-centered gene networks at 24h. Conclusions Our data show that both CBHA and TSA induced similar, but not identical, time-dependent, gene networks in H9c2 cardiac myocytes. Initially, both HDACIs impinged on numerous genes associated with adipokine signaling, intracellular metabolism and energetics, and cell cycle. A continued exposure to either CBHA or TSA led to the emergence of a number of apoptosis- and inflammation-specific gene networks that were apparently suppressed by both HDACIs. Based on these data we posit that the anti-inflammatory and anti-proliferative actions of HDACIs are myocyte-intrinsic. These findings advance our understanding of the mechanisms of actions of HDACIs on cardiac myocytes and reveal potential signaling pathways that may be targeted therapeutically.



Effects of KR31378, a novel ATP-sensitive potassium channel activator, on hypertrophy of H9c2 cells and on cardiac dysfunction in rats with congestive heart failure  

Microsoft Academic Search

The present study was performed to evaluate the effects of (2S, 3S, 4R)-N?-cyano-N-(6-amino-3, 4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran-4yl)-N?-benzylguanidine (KR-31378), a novel mitochondrial ATP-sensitive potassium channel activator, on hypertrophy of H9c2 cells and on cardiac dysfunction in rats with congestive heart failure. In rat heart-derived H9c2 cells treated with hypertrophic agonists, such as angiotensin II, phenylephrine, isoproterenol, and urotensin II, cell size was significantly increased

Geum Shil Hwang; Kwang-Seok Oh; Hyun-Na Koo; Ho Won Seo; Kwan-Hee You; Byung Ho Lee



Inhibitory effects of rosmarinic acid on adriamycin-induced apoptosis in H9c2 cardiac muscle cells by inhibiting reactive oxygen species and the activations of c-Jun N-terminal kinase and extracellular signal-regulated kinase  

Microsoft Academic Search

Rosmarinic acid (RA) is a naturally occurring polyphenolic and is found in several herbs in the Lamiaceae family, such as, Perilla frutescens. ADR is a potent anti-tumor drug, but is unfortunately potently cardiotoxic. This study was undertaken to investigate the inhibitory effect of RA on ADR-induced apoptosis in H9c2 cardiac muscle cells at a mechanistic level. In vitro, ADR significantly

Do-Sung Kim; Hyung-Ryong Kim; Eun-Rhan Woo; Seong-Tshool Hong; Han-Jung Chae; Soo-Wan Chae



Differentiation-dependent doxorubicin toxicity on H9c2 cardiomyoblasts.  


A characteristic component of the anti-neoplastic doxorubicin (DOX)-induced cardiac toxicity is the delayed and persistent toxicity, with cancer childhood survivors developing cardiac failure later in life. The mechanisms behind this persistent toxicity are unknown, although one of the consequences of early childhood treatment with DOX is a specific removal of cardiac progenitor cells. DOX treatment may be more toxic to undifferentiated muscle cells, contributing to impaired cardiac development and toxicity persistence. H9c2 myoblasts, a rat embryonic cell line, which has the ability to differentiate into a skeletal or cardiac muscle phenotype, can be instrumental in understanding DOX cytotoxicity in different differentiation stages. H9c2 cell differentiation results in decreased cell proliferation and increased expression of a differentiated muscle marker. Differentiated H9c2 cells accumulated more DOX and were more susceptible to DOX-induced cytotoxicity. Differentiated cells had increased levels of mitochondrial superoxide dismutase and Bcl-xL, an anti-apoptotic protein. Of critical importance for the mechanisms of DOX toxicity, p53 appeared to be equally activated regardless of the differentiation state. We suggest that although more differentiated H9c2 muscle cells appear to have more basal mechanisms that would predict higher protection, DOX toxicity is higher in the differentiated population. The results are instrumental in the understanding of stress responses of this specific cell line in different differentiation stages to the cardiotoxicity caused by anthracyclines. PMID:22744233

Branco, Ana F; Sampaio, Susana F; Moreira, Ana C; Holy, Jon; Wallace, Kendall B; Baldeiras, Ines; Oliveira, Paulo J; Sardão, Vilma A



KR32570, a novel Na + \\/ H + exchanger-1 inhibitor, attenuates hypoxia-induced cell death through inhibition of intracellular Ca 2+ overload and mitochondrial death pathway in H9c2 cells  

Microsoft Academic Search

A novel Na+\\/H+ exchanger-1 (NHE-1) inhibitor [5-(2-methoxy-5-chloro-5-phenyl)furan-2-ylcarbonyl]guanidine (KR-32570) has been previously demonstrated to elicit cardioprotective effect against ischemic injury in rat heart. In the present study, we examined the effects of KR-32570 on cell death induced by hypoxic insult in heart-derived H9c2 cells. Treatment with KR-32570 (1–10 ?M) significantly reduced hypoxia-induced necrotic cell death (lactate dehydrogenase release) and apoptotic cell

Mi Jeong Kim; Chang-Hyun Moon; Mi-Young Kim; Sunkyung Lee; Kyu Yang Yi; Sung Eun Yoo; Soo Hwan Lee; Eun Joo Baik; Yi-Sook Jung



Exogenous hydrogen sulfide protects H9c2 cardiac cells against high glucose-induced injury by inhibiting the activities of the p38 MAPK and ERK1/2 pathways.  


Hyperglycemia is a risk factor for the development of diabetic cardiovascular complications, which are associated with the activation of the mitogen-activated protein kinase (MAPK) signaling pathway. In this study, we demonstrate the inhibitory effects of exogenous hydrogen sulfide (H2S) on the activation of the MAPK pathway. The aim of the present study was to determine whether exogenous H2S prevents high glucose (HG)-induced injury by inhibiting the activation of the p38 MAPK and extracellular signal-regulated kinase (ERK)1/2 (members of MAPK) pathways in cardiomyoblasts (H9c2 cells). The findings of the present study demonstrated that the treatment of H9c2 cells with HG (35 mM glucose) for 24 h not only significantly induced injury, including cytotoxicity, apoptosis, overproduction of reactive oxygen species (ROS) and the loss of mitochondrial membrane potential (MMP), but also upregulated the expression levels of phosphorylated (p)-p38 MAPK and p-ERK1/2. The increased expression levels of p-p38 MAPK and p-ERK1/2 were markedly reduced by pre-treatment of the H9c2 cells with 400 µM sodium hydrogen sulfide (NaHS; a donor of H2S) prior to exposure to 35 mM glucose. Importantly, pre-treatment of the cells with 400 µM NaHS or 3 µM SB203580 (a selective inhibitor of p38 MAPK) or 15 µM U0126 (a selective inhibitor of ERK1/2) attenuated the HG-induced cardiomyocyte injury, leading to an increase in cell viability and a decrease in the number of apoptotic cells, preventing ROS generation, as well as the loss of MMP. In addition, pre-treatment of the cells with 1,000 µM N?acetyl?L?cysteine (a ROS scavenger) prior to exposure to HG ameliorated the HG-induced cytotoxicity. Taken together, the data from the present study demonstrate for the first time, to our knowledge, that exogenous H2S exerts a protective effect against HG?induced injury by inhibiting the activation of the p38 MAPK and ERK1/2 pathways and preventing oxidative stress in H9c2 cells. PMID:23912965

Xu, Wenming; Wu, Wen; Chen, Jingfu; Guo, Runmin; Lin, Jiancong; Liao, Xinxue; Feng, Jianqiang



HSP27 regulates p53 transcriptional activity in doxorubicin-treated fibroblasts and cardiac H9c2 cells: p21 upregulation and G2/M phase cell cycle arrest.  


Treatment of cancer patients with anthracyclin-based chemotherapeutic drugs induces congestive heart failure by a mechanism involving p53. However, it is not known how p53 aggravates doxorubicin (Dox)-induced toxicity in the heart. On the basis of in vitro acute toxicity assay using heat shock factor-1 (HSF-1) wild-type (HSF-1(+/+)) and HSF-1-knockout (HSF-1(-/-)) mouse embryonic fibroblasts and neonatal rat cardiomyocyte-derived H9c2 cells, we demonstrate a novel mechanism whereby heat shock protein 27 (HSP27) regulates transcriptional activity of p53 in Dox-treated cells. Inhibition of p53 by pifithrin-alpha (PFT-alpha) provided different levels of protection from Dox that correlate with HSP27 levels in these cells. In HSF-1(+/+) cells, PFT-alpha attenuated Dox-induced toxicity. However, in HSF-1(-/-) cells (which express a very low level of HSP27 compared with HSF-1(+/+) cells), there was no such attenuation, indicating an important role of HSP27 in p53-dependent cell death. On the other hand, immunoprecipitation of p53 was found to coimmunoprecipitate HSP27 and vice versa (confirmed by Western blotting and matrix-assisted laser desorption/ionization time of flight), demonstrating HSP27 binding to p53 in Dox-treated cells. Moreover, upregulation of p21 was observed in HSF-1(+/+) and H9c2 cells, indicating that HSP27 binding transactivates p53 and enhances transcription of p21 in response to Dox treatment. Further analysis with flow cytometry showed that increased expression of p21 results in G(2)/M phase cell cycle arrest in Dox-treated cells. Overall, HSP27 binding to p53 attenuated the cellular toxicity by upregulating p21 and prevented cell death. PMID:18263706

Venkatakrishnan, C D; Dunsmore, Kathy; Wong, Hector; Roy, Sashwathi; Sen, Chandan K; Wani, Altaf; Zweier, Jay L; Ilangovan, Govindasamy



Arginine vasopressin enhances cell survival via a G protein-coupled receptor kinase 2/?-arrestin1/extracellular-regulated kinase 1/2-dependent pathway in H9c2 cells.  


Circulating levels of arginine vasopressin (AVP) are elevated during hypovolemia and during cardiac stress. AVP activates arginine vasopressin type 1A (V(1A))/G?(q)-coupled receptors in the heart and vasculature and V(2)/G?(s)-coupled receptors in the kidney. However, little is known regarding the signaling pathways that influence the effects of V(1A) receptor (V(1A)R) activation during cellular injury. Using hypoxia-reoxygenation (H/R) as a cell injury model, we evaluated cell survival and caspase 3/7 activity in H9c2 myoblasts after treatment with AVP. Pretreatment of H9c2 cells with AVP significantly reduced H/R-induced cell death and caspase 3/7 activity, effects that were blocked via both selective V(1A)R inhibition and mitogen-activated protein kinase (MEK1/2) inhibition. AVP increased extracellular-regulated kinase 1/2 (ERK1/2) phosphorylation in a concentration-dependent manner that was sensitive to MEK1/2 inhibition and V(1A)R inhibition, but not V(1B)R or V(2)R inhibition. Discrete elements of the V(1A)/G?(q)-protein kinase C (PKC) and V(1A)/G protein-coupled receptor kinase (GRK)/?-arrestin signaling cascades were inhibited to dissect the pathways responsible for the protective effects of V(1A)R signaling: G?(q) (overexpression of Gq-I-ires-green fluorescent protein), PKC (administration of Ro 31-82425; 2-[8-(aminomethyl)-6,7,8,9-tetrahydropyrido[1,2-a]indol-3-yl]-3-(1-methyl-1H-indol-3-yl)maleimide, HCl, bisindolylmaleimide X, HCl), GRK2 [C-terminal GRK2 peptide overexpression and small interfering RNA (siRNA) knockdown], GRK5 (siRNA knockdown), and ?-arrestin1 (siRNA knockdown). These studies demonstrated that both G?(q)/PKC- and GRK2/?-arrestin1-dependent V(1A)R signaling were capable of inducing ERK1/2 phosphorylation in response to AVP stimulation. However, AVP-mediated protection against H/R was elicited only via GRK2- and ?-arrestin1-dependent signaling. These results suggest that activation of the V(1A)R in H9c2 cells mediates protective signaling via a GRK2/?-arrestin1/ERK1/2-dependent mechanism that leads to decreased caspase 3/7 activity and enhanced survival under conditions of ischemic stress. PMID:23690069

Zhu, Weizhong; Tilley, Douglas G; Myers, Valerie D; Coleman, Ryan C; Feldman, Arthur M



Mitochondrial membrane potential measurement of H9c2 cells grown in high-glucose and galactose-containing media does not provide additional predictivity towards mitochondrial assessment.  


Drug-induced mitochondrial toxicity is a contributing factor to many organ toxicities. The fact that some, but not all members of a particular drug class can induce mitochondrial dysfunction has necessitated the need for predictive screens within the drug development process. One of these screens is a cell viability assay done in two types of media, one containing high-glucose, the other, galactose. Since galactose-grown cells are more susceptible to mitochondrial toxicants than high-glucose-grown cells, this assay distinguishes compounds that cause toxicity primarily through mitochondrial targets from those that cause multifactorial toxicity. However, the assay does not show if compounds that cause multifactorial toxicity cause impairment on mitochondria. To address this problem, we investigated if multiplexing the assay with mitochondrial membrane potential measurements using the fluorescent dye, JC-1, could provide further information. We tested 28 drugs in the multiplexed assay and found that, although mitochondrial toxicants could be detected, no additional information was revealed about compounds that caused multifactorial toxicity. Hence, measurements with JC-1 did not provide additional information beyond what was detected using the cell viability assay. We conclude that even though the multiplexed assay is useful for HTS applications, it provides no additional value over the high-glucose-galactose cell viability assay. PMID:21126567

Rana, Payal; Nadanaciva, Sashi; Will, Yvonne



Ursolic-Acid-Enriched Herba Cynomorii Extract Protects against Oxidant Injury in H9c2 Cells and Rat Myocardium by Increasing Mitochondrial ATP Generation Capacity and Enhancing Cellular Glutathione Redox Cycling, Possibly through Mitochondrial Uncoupling  

PubMed Central

Mitochondrial decay is considered to be a major contributor to aging-related diseases, including neurodegenerative diseases, cardiovascular disorders, and certain metabolic diseases. Therefore, the maintenance of mitochondrial functional capacity and antioxidant status should play an essential role in preventive health. Herba Cynomorii, which is one of the most potent “Yang-invigorating” Chinese tonic herbs, was found to increase mitochondrial ATP generation capacity (ATP-GC) in rat hearts ex vivo. In the present study, we demonstrated that HCY2, an active fraction of Herba Cynomorii, and its major ingredient ursolic acid (UA) could protect against hypoxia/reoxygenation-induced cell apoptosis in H9c2 cells in vitro and also against ischemia/reperfusion-induced injury in rat hearts ex vivo. The cardioprotection was associated with an increase in ATP-GC and an enhancement of glutathione redox cycling. The results suggest that UA may be one of the active ingredients responsible for the cardioprotection afforded by Herba Cynomorii, and this effect may be mediated, at least in part, by enhancement of mitochondrial functional capacity and antioxidant status, possibly through the induction of mitochondrial uncoupling.

Ko, Kam Ming



Vasopressin-induced hypertrophy in H9c2 heart-derived myocytes  

Microsoft Academic Search

Protein synthesis in H9c2 heart-derived myocytes responds biphasically to arginine vasopressin (1 ?M). An initial 50% inhibition attributable to Ca2+ mobilization from the sarcoplasmic\\/endoplasmic reticulum is followed by a recovery that subsequently converts to a 1.5-fold stimulation. This study was undertaken to ascertain whether vasopressin programs H9c2 cells to undergo hypertrophy or to proliferate and whether early translational inhibition is

Margaret A. Brostrom; Barbara A. Reilly; Frank J. Wilson; Charles O. Brostrom



Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets  

Microsoft Academic Search

Doxorubicin (Dox) is a very potent antineoplastic agent used against several types of cancer, despite a cumulative cardiomyopathy\\u000a that reduces the therapeutic index for treatment. H9c2 myoblast cells have been used as an in vitro model to study biochemical\\u000a alterations induced by Dox treatment on cardiomyocyte cells. Despite the extensive work already published, few data are available\\u000a regarding morphological alterations

Vilma A. Sardão; Paulo J. Oliveira; Jon Holy; Catarina R. Oliveira; Kendall B. Wallace



Involvement of JNKs and p38MAPK\\/MSK1 pathways in H 2O 2-induced upregulation of heme oxygenase-1 mRNA in H9c2 cells  

Microsoft Academic Search

One of the most important challenges that cardiomyocytes experience is an increase in the levels of reactive oxygen species (ROS), i.e., during ischemia, reperfusion as well as in the failing myocardium. HOX-1 has been found to protect cells and tissues against oxidative damage; therefore, we decided to study the signalling cascades involved in its transcriptional regulation. HOX-1 mRNA levels were

Ioanna-Katerina S. Aggeli; Catherine Gaitanaki; Isidoros Beis



Pro-survival effect of Dock180 overexpression on rat-derived H9C2 cardiomyocytes  

PubMed Central

Background Integrin ?1 subunit and its downstream molecule, focal adhesion kinase (FAK), have been demonstrated to be indispensible to the promotion of cell proliferation and survival and anti-apoptosis in cardiomyocytes via activation of their downstream pro-survival signaling molecule, AKT. As a component of the integrin pathway, Dock180 (dedicator of cytokinesis 1) protein is also thought to be involved in the promotion of cell proliferation and survival and anti-apoptosis in the H9C2 cardiomyocytes. Material/Methods Rat-derived H9C2 cardiomyocytes were transfected with pCXN2-flag-hDock180, a human Dock180 overexpression eukaryotic recombinant plasmid. The rat and human Dock180 mRNA and protein expression, apoptosis and cell proliferation and survival were analyzed in the H9C2 cardiomyocytes treated with either hypoxia/reoxygenation (H/R) or not, respectively. Results Human Dock180 mRNA overexpression could significantly increase the Dock180 protein expression in the H9C2 cardiomyocytes, no matter whether treated with H/R or not. Dock180 overexpression could promote the cell proliferation and survival and anti-apoptosis, and relieve the cell proliferative and survival inhibition and apoptosis induced by H/R in the H9C2 cardiomyocytes via activation of its downstream pro-survival signaling molecule AKT. Conclusions Dock180 could act as a pro-survival molecule in H9C2 cardiomyocytes via activation of its downstream pro-survival signaling molecule, AKT.

Yan, An; Li, Gang; Zhang, Xu; Zhu, Bingbao; Linghu, Hua



Curcumin Suppresses Gelatinase B Mediated Norepinephrine Induced Stress in H9c2 Cardiomyocytes  

PubMed Central

Background Extracellular matrix (ECM) remodeling facilitates biomechanical signals in response to abnormal physiological conditions. This process is witnessed as one of the major effects of the stress imposed by catecholamines, such as epinephrine and norepinephrine (NE), on cardiac muscle cells. Matrix metalloproteinases (MMPs) are the key proteases involved in degradation of the ECM in heart. Objectives The present study focuses on studying the effect of curcumin on Gelatinase B (MMP-9), an ECM remodeling regulatory enzyme, in NE-induced cardiac stress. Curcumin, a bioactive polyphenol found in the spice turmeric, has been studied for its multi-fold beneficial properties. This study focuses on investigating the role of curcumin as a cardio-protectant. Methods H9c2 cardiomyocytes were subjected to NE and curcumin treatments to study the response in stress conditions. Effect on total collagen content was studied using Picrosirus red staining. Gelatinase B activity was assessed through Gel-Diffusion Assay and Zymographic techniques. RT-PCR, Western Blotting and Immunocytochemistry were performed to study effect on expression of gelatinase B. Further, the effect of curcumin on the localization of NF-?B, known to regulate gelatinase B, was also examined. Results Curcumin suppressed the increase in the total collagen content under hypertrophic stress and was found to inhibit the in-gel and in-situ gelatinolytic activity of gelatinase B. Moreover, it was found to suppress the mRNA and protein expression of gelatinase B. Conclusions The study provides an evidence for an overall inhibitory effect of curcumin on Gelatinase B in NE-induced hypertrophic stress in H9c2 cardiomyocytes which may contribute in the prevention of ECM remodeling.

Kohli, Shrey; Chhabra, Aastha; Jaiswal, Astha; Rustagi, Yashika; Sharma, Manish; Rani, Vibha



Phosphatidylinositol 3Kinase Regulates Differentiation of H9c2 Cardiomyoblasts Mainly through the Protein Kinase B\\/Akt-Independent Pathway  

Microsoft Academic Search

Phosphatidylinositol 3-kinase (PI3-kinase) is known to be a crucial regulator of muscle differentiation. However, its downstream pathway for this function is quite obscure. In this experiment we demonstrated the regulatory mechanism of the differentiation of H9c2 cardiomyoblasts, focusing on PI3-kinase, protein kinase B\\/Akt (PKB\\/Akt) and p42\\/44 mitogen-activated protein kinase (p42\\/44 MAPK). When H9c2 cells stably transfected with a constitutively active

Joung Mok Kim; Moon-young Yoon; Jayoung Kim; Sam Soo Kim; Insug Kang; Joohun Ha; Sung Soo Kim



Modulation of the caveolin-3 and Akt status in caveolae by insulin resistance in H9c2 cardiomyoblasts  

Microsoft Academic Search

We investigated glucose uptake and the trans- location of Akt and caveolin-3 in response to insulin in H9c2 cardiomyoblasts exposed to an experimental insulin resistance condition of 100 nM insulin in a 25 mM glucose containing media for 24 h. The cells under the insulin resistance condition exhibited a decrease in insulin-sti- mulated 2-deoxy( 3 H)glucose uptake as compared to

Hyunil Ha; Yunbae Pak


Signal Transduction of Arginine Vasopressin-Induced Arachidonic Acid Release in H9c2 Cardiac Myoblasts: Role of Ca2+ and the Protein Kinase C-Dependent Activation of p42 Mitogen-Activated Protein Kinase  

Microsoft Academic Search

The mechanism of arginine vasopressin (AVP)-induced arachi- donic acid (AA) release was examined in the cardiac myoblast cell line, H9c2. Stimulation of cells with AVP induced dose-dependent AA release, and this effect was completely inhibited by the V1 receptor antagonist, d(CH)5(Tyr(Me) 2 )AVP. AVP also produced dose-depen- dent stimulation of inositol phosphate formation; this was not affected by pertussis toxin,




C3G overexpression promotes the survival of rat-derived H9C2 cardiomyocytes by p-ERK1/2.  


Integrin ?1 subunit and its downstream molecules, such as integrin-linked kinase and focal adhesion kinase, are imperative for promotion of cell proliferation, survival and anti-apoptosis in cardiomyocytes by activation of their downstream pro-survival signalling molecules, such as the phosphorylated extracellular signal-regulated kinase1/2 (p-ERK1/2). As a component of the integrin pathway, C3G (Crk-SH3 domain guanine nucleotide exchange factor) protein may be involved in the promotion of cell proliferation and survival and anti-apoptosis in the H9C2 cardiomyocytes. Rat-derived H9C2 cardiomyocytes were transfected with pCXN2-flag-hC3G, a human C3G overexpression eukaryotic recombinant plasmid. Apoptosis, cell proliferation and survival were analysed in the H9C2 cardiomyocytes either treated with hypoxia/reoxygenation (H/R). Human C3G mRNA overexpression significantly elevated C3G protein expression in H9C2 cardiomyocytes whether treated with H/R or not. C3G overexpression promoted proliferation and survival and anti-apoptosis, and attenuated the proliferative and survival inhibition, and apoptosis induced by H/R by activation of its downstream pro-survival signalling molecule, p-ERK1/2. The results suggest that C3G acts as a pro-survival molecule in H9C2 cardiomyocytes by activation of p-ERK1/2. PMID:23686869

Zhang, Xu; Li, Gang; Zhang, Lei; Yang, Dongyan; Zhang, Zhisheng; Yan, An; Linghu, Hua



Mitochondrial disruption occurs downstream from ?-adrenergic overactivation by isoproterenol in differentiated, but not undifferentiated H9c2 cardiomyoblasts: Differential activation of stress and survival pathways.  


?-Adrenergic receptor stimulation plays an important role in cardiomyocyte stress responses, which may result in apoptosis and cardiovascular degeneration. We previously demonstrated that toxicity of the ?-adrenergic agonist isoproterenol on H9c2 cardiomyoblasts depends on the stage of cell differentiation. We now investigate ?-adrenergic receptor downstream signaling pathways and stress responses that explain the impact of muscle cell differentiation on hyper-?-adrenergic stimulation-induced cytotoxicity. When incubated with isoproterenol, differentiated H9c2 muscle cells have increased cytosolic calcium, cyclic-adenosine monophosphate content and oxidative stress, as well as mitochondrial depolarization, increased superoxide anion, loss of subunits from the mitochondrial respiratory chain, decreased Bcl-xL content, increased p53 and phosphorylated-p66Shc as well as activated caspase-3. Undifferentiated H9c2 cells incubated with isoproterenol showed increased Bcl-xL protein and increased superoxide dismutase 2 which may act as protective mechanisms. We conclude that the differentiation of H9c2 is associated with differential regulation of stress responses, which impact the toxicity of several agents, namely those acting through ?-adrenergic receptors and resulting in mitochondrial disruption in differentiated cells only. PMID:23958426

Branco, Ana F; Sampaio, Susana F; Wieckowski, Mariusz R; Sardão, Vilma A; Oliveira, Paulo J



H9c2 cardiac myoblasts undergo apoptosis in a model of ischemia consisting of serum deprivation and hypoxia: inhibition by PMA  

Microsoft Academic Search

Cardiac myocytes undergo apoptosis under condition of ischemia. Little is known, however, about the molecular pathways that mediate this response. We show that serum deprivation and hypoxia, components of ischemia in vivo, resulted in apoptosis of rat ventricular myoblast cells H9c2. Hypoxia alone did not induce significant apoptosis for at least 48 h, but largely increased the proapoptotic action of

Francesca Bonavita; Claudio Stefanelli; Emanuele Giordano; Marta Columbaro; Annalisa Facchini; Francesca Bonafè; Claudio Marcello Caldarera; Carlo Guarnieri



Activin receptor-like kinase 7 mediates high glucose-induced H9c2 cardiomyoblast apoptosis through activation of Smad2/3.  


Cardiomyocyte apoptosis is an important pathological change of diabetic cardiomyopathy. How the elevated glucose levels cause cell apoptosis remains unknown. The aim of our study was to investigate whether activin receptor-like kinase 7 (ALK7)-Smad2/3 signaling pathway plays an important role in high glucose-induced cardiomyocyte apoptosis. H9c2 cardiomyoblasts and neonatal rat cardiomyocytes were treated with 33mmol/l glucose. The expression of ALK7, Smad2 and Smad3 were inhibited by small interfering RNA respectively. The level of ALK7, total Smad2/3, phosphorylated Smad2/3, B-cell lymphoma-2 (Bcl-2) and cleaved Caspase3 were evaluated using western blot. The apoptosis rate was detected by flow cytometer. High glucose treatment caused the apoptosis of H9c2 cardiomyocyte and the inhibition of Smad2 or Smad3 attenuated this apoptosis. ALK7 existed in both H9c2 cardiomyoblasts and neonatal rat cardiomyocytes and high ambient glucose upregulated its expression. The increased expression level of cleaved Caspase3 and apoptosis rate and decreased expression of Bcl-2 were reversed after ALK7 was inhibited. The expression of phosphorylated Smad2/3 also decreased after the knockdown of ALK7. Our findings suggest that ALK7 mediates high ambient glucose-induced H9c2 cardiomyoblasts apoptosis through the activation of Smad2/3. PMID:23830891

Liu, Lin; Ding, Wen-yuan; Zhao, Jing; Wang, Zhi-hao; Zhong, Ming; Zhang, Wei; Chen, Yu-guo; Zhang, Yun; Li, Li; Tang, Meng-xiong



Cardioprotective role of Syzygium cumini against glucose-induced oxidative stress in H9C2 cardiac myocytes.  


Diabetic patients are known to have an independent risk of cardiomyopathy. Hyperglycemia leads to upregulation of reactive oxygen species (ROS) that may contribute to diabetic cardiomyopathy. Thus, agents that suppress glucose-induced intracellular ROS levels can have therapeutic potential against diabetic cardiomyopathy. Syzygium cumini is well known for its anti-diabetic potential, but its cardioprotective properties have not been evaluated yet. The aim of the present study is to analyze cardioprotective properties of methanolic seed extract (MSE) of S. cumini in diabetic in vitro conditions. ROS scavenging activity of MSE was studied in glucose-stressed H9C2 cardiac myoblasts after optimizing the safe dose of glucose and MSE by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide. 2',7'-dichlorfluorescein diacetate staining and Fluorescence-activated cell sorting analysis confirmed the suppression of ROS production by MSE in glucose-induced cells. The intracellular NO and H2O2 radical-scavenging activity of MSE was found to be significantly high in glucose-induced cells. Exposure of glucose-stressed H9C2 cells to MSE showed decline in the activity of catalase and superoxide dismutase enzymes and collagen content. 4',6-diamidino-2-phenylindole, propidium iodide and 10-N-nonyl-3,6-bis (dimethylamino) acridine staining revealed that MSE protects myocardial cells from glucose-induced stress. Taken together, our findings revealed that the well-known anti-diabetic S. cumini can also protect the cardiac cells from glucose-induced stress. PMID:23512199

Atale, Neha; Chakraborty, Mainak; Mohanty, Sujata; Bhattacharya, Susinjan; Nigam, Darshika; Sharma, Manish; Rani, Vibha



Cytoprotective effect of anthocyanins against doxorubicin-induced toxicity in H9c2 cardiomyocytes in relation to their antioxidant activities  

Microsoft Academic Search

The effect of six anthocyanidins and seven anthocyanins against doxorubicin (Dox)-induced cardiotoxicity in relation to their antioxidant properties was investigated in H9c2 cardiomyocytes. The exposure to Dox, a highly effective cytotoxic agent against cancer cells, induced significant cell death, intracellular reactive oxygen species (ROS), and lipid peroxidation in non-tumorigenic cardiac cell culture. All anthocyanidins (50 and\\/or 100?M) significantly increased cell

Eun Hye Choi; Hyun-Joo Chang; Jae Young Cho; Hyang Sook Chun



Gui-ling-gao, a traditional Chinese functional food, prevents oxidative stress-induced apoptosis in H9c2 cardiomyocytes.  


Functional foods have become an increasingly popular alternative to prevent diseases and maintain body health status. Gui-ling-gao (GLG, also known as turtle jelly) is a well-known traditional functional food popular in Southern China and Hong Kong. This study aimed to investigate the antioxidative and anti-apoptotic effects of GLG, a traditional Chinese functional food, on preventing oxidative stress-induced injury in H9c2 cardiomyocytes. In this study, the antioxidative capacities of GLG were measured by using both a cell-free assay [2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl assay] and biological methods [2,2'-azobis(2-amidinopropane)-induced haemolysis assay and H(2)O(2)-induced cell damage on H9c2 cardiomyocytes]. Additionally, the total phenolic content was measured using the Folin-Ciocalteu method. Furthermore, the anti-apoptotic effect of GLG was evaluated by nuclear staining and a DNA fragmentation assay. GLG was found to have good antioxidant activities and high total phenolic content. In H(2)O(2)-induced cell damage on H9c2 cells, GLG was demonstrated to ameliorate the apoptotic effects, such as nuclear condensations, increased intracellular caspase-3 activity and inter-nucleosomal DNA cleavage, induced by H(2)O(2). The present study demonstrated for the first time that GLG possesses anti-apoptotic potential in vitro and this effect may be mediated, in part, by its antioxidative function. Additionally, the antioxidative capacities of GLG were proved both chemically and biologically. This study provides scientific evidence to prove the anecdotal health-beneficial claim that the consumption of GLG could help the body to handle endogenous toxicants such as free radicals. PMID:23467630

Li, Fan; Wu, Jian-Hong; Wang, Qing-Hua; Shu, Yuan-Lan; Wan, Chun-Wai; Chan, Chi-On; Kam-Wah Mok, Daniel; Chan, Shun-Wan



Hexarelin protects H9c2 cardiomyocytes from doxorubicin-induced cell death  

Microsoft Academic Search

Growth hormone secretagogues (GHSs) are synthetic peptidyl and nonpeptidyl molecules that possess strong growth hormone-releasing\\u000a activity acting on specific pituitary and hypothalamic receptor subtypes. Differently from nonpeptidyl GHSs, peptidyl molecules\\u000a such as hexarelin, a hexapeptide, possess specific high-affinity binding sites in animal and human heart and, after prolonged\\u000a treatment, protect rats in vivo from ischemiainduced myocardial damage. To verify the

Nicoletta Filigheddu; Alberto Fubini; Gianluca Baldanzi; Santina Cutrupi; Corrado Ghè; Filomena Catapano; Fabio Broglio; Amalia Bosia; Mauro Papotti; Giampiero Muccioli; Ezio Ghigo; Romano Deghenghi; Andrea Graziani



Glutathione-S-transferase overexpression protects against anthracycline-induced H9C2 cell death  

Microsoft Academic Search

Abstract Anthracyclines (AC) are anti-tumor antibiotics with significant activity against solid and hematologic malignancies. One problem preventing more widespread use has been the development,of cardiac toxicity. Experimental evidence supports oxidant stress as an important trigger and\\/or mediator of AC-induced cardiotoxicity (ACT). Therefore, reducing oxidant stress should be protective against ACT. To determine whether antioxidant protein overexpression can reduce ACT, we

Thomas L'Ecuyer



A Physiologically Relevant Hyperthermia Selectively Activates Constitutive hsp70 in H9c2 Cardiac Myoblasts and Confers Oxidative Protection  

Microsoft Academic Search

Whole body hyperthermia (42–43°C for 15–20 min) elicits the formation of heat shock proteins (hsps) and improves cardiac recovery from subsequent ischemia\\/reperfusion. However, the beneficial effects of this response are compromised by initial tissue injury, which limits its clinical applicability. Using a simplified myocardial model (rat heart-derived H9c2 myoblasts) a hypothesis was tested that chronic, mild hyperthermia is as effective

Ching-Yuan Su; Kowit-Yu Chong; JianXiong Chen; Stefan Ryter; Romesh Khardori; Chen-Ching Lai



Doxorubicin-induced mitochondrial dysfunction is secondary to nuclear p53 activation in H9c2 cardiomyoblasts  

Microsoft Academic Search

Purpose  Doxorubicin (DOX) is a widely prescribed chemotherapeutic. The hypothesis for the present study is that DOX-induced myocyte\\u000a apoptosis involves mitochondrial dysfunction that is a consequence of nuclear DOX effects.\\u000a \\u000a \\u000a \\u000a Methods  H9c2 myoblasts were incubated with 0, 0.5 and 1 ?M DOX and nuclear and mitochondrial alterations were determined.\\u000a \\u000a \\u000a \\u000a Results  Doxorubicin accumulation in the nucleus was detected after 3 h treatment, followed by an increase

Vilma A. Sardão; Paulo J. Oliveira; Jon Holy; Catarina R. Oliveira; Kendall B. Wallace



Silencing of angiotensin?converting enzyme by RNA interference prevents H9c2 cardiomyocytes from apoptosis induced by anoxia/reoxygenation through regulation of the intracellular renin-angiotensin system.  


Inhibition of the angiotensin?converting enzyme (ACE) attenuated apoptotic cardiomyocytes induced by ischemic reperfusion (I/R). However, it is difficult to evaluate the effects of inhibition of the intracellular ACE in vivo. The objective of this study was to determine whether the apoptosis in H9c2 cardiomyocytes following anoxia/reoxygenation (A/R) would be improved by the silencing of intracellular ACE by RNA interference (RNAi). H9c2 cardiomyocytes were subjected to A/R 48 h following transfection with ACE-shRNA plasmid. The results showed that the gene silencing of intracellular ACE significantly inhibited the decrease of cell viability and the increase of apoptotic H9c2 cardiomyocytes undergoing A/R. Additionally, the gene silencing of intracellular ACE significantly promoted the expression of ACE2, decreased caspase?3 activity and Bax levels, and enhanced the expression of Bcl?2 in H9c2 cardiomyocytes subjected to A/R. The results suggest that the gene silencing of intracellular ACE holds great potential in the treatment of cardiomyocyte apoptosis following I/R injury through the regulation of the intracellular renin?angiotensin system, thereby regulating the intrinsic pathway of apoptosis. PMID:24126381

Wan, Weiguo; Jiang, Xuejun; Li, Xiaoyan; Zhang, Cui; Yi, Xin



Insulin-like growth factor-1 protects H9c2 cardiac myoblasts from oxidative stress-induced apoptosis via phosphatidylinositol 3-kinase and extracellular signal-regulated kinase pathways  

Microsoft Academic Search

Oxidative stress plays a critical role in cardiac injuries during ischemia\\/reperfusion. Insulin-like growth factor-1 (IGF-1) promotes cell survival in a number of cell types, but the effect of IGF-1 on the oxidative stress has not been elucidated in cardiac muscle cells. Therefore, we examined the role of IGF-1 signaling pathway in cell survival against H2O2-induced apoptosis in H9c2 cardiac myoblasts.

Feng Hong; Si Joong Kwon; Bong Sook Jhun; Sung Soo Kim; Joohun Ha; Soo-Ja Kim; Nak Won Sohn; Chulhun Kang; Insug Kang



Cytochrome P-450-catalyzed reactive oxygen species production mediates the (-)schisandrin B-induced glutathione and heat shock responses in H9c2 cardiomyocytes  

PubMed Central

Objective: Schisandrin B (Sch B) is the most abundant, active dibenzocyclooctadiene derivative isolated from the fruit of Schisandra chinensis (Turcz) Baillon (Schisandraceae). (–)Sch B was found to be the most potent stereoisomer of Sch B in producing cytoprotective action in H9c2 cardiomyocytes. The elucidation of biochemical mechanism underlying the cytoprotection of (–)Sch B has attracted much interest in the area of preventive medicine. Here, we examined whether the (–)Sch B-induced enhancement of glutathione antioxidant and heat shock responses and the associated cytoprotection against hypoxia/reoxygenation-induced apoptosis are mediated by reactive oxygen species (ROS) arising from cytochrome P-450 (CYP)-catalyzed metabolism of (–)Sch B in H9c2 cardiomyocytes. Materials and Methods: The effects of CYP inhibitor (1-aminobenzotriazole, ABT) and antioxidant (dimethylthiouracil, DMTU) on (–)Sch B-induced ROS production and associated increases in cellular-reduced glutathione (GSH) level as well as heat shock protein (Hsp) 25/70 production were investigated in H9c2 cardiomyocytes. The (–)Sch B-induced ROS generation was monitored with or without ABT/DMTU for 6 h in situ, while (–)Sch B-induced cellular GSH level and Hsp 25/70 production, as well as cytoprotection were measured at 16 h post-(–)Sch B exposure. Results: The results indicated that (–)Sch B caused a dose-dependent increase in ROS production in H9c2 cardiomyocytes, which was completely suppressed by pre- and co-treatment with ABT or DTMU. The incubation with (–)Sch B for 6 h caused dose-dependent increases in cellular GSH level and Hsp 25/70 production, as well as protection against hypoxia/reoxygenation-induced apoptosis at 16-h post-drug exposure in H9c2 cardiomyocytes. All these cellular responses were abrogated by treatment with ABT or DMTU. Conclusion: The results suggest that ROS arising from the CYP-catalyzed metabolism of (–)Sch B elicit glutathione antioxidant and heat shock responses, thereby protecting against oxidant-induced apoptosis in H9c2 cardiomyocytes.

Chen, Na; Chiu, Po Yee; Leung, Hoi Yan; Ko, Kam Ming



Hexane/ethanol extract of Glycyrrhiza uralensis licorice suppresses doxorubicin-induced apoptosis in H9c2 rat cardiac myoblasts.  


Doxorubicin (DOX) is an anthracycline antibiotic, and has been recognized as one of the most effective anti-neoplastic agents in cancer chemotherapy. However, its usefulness is limited by its profound cardiotoxicity. Licorice is one of the most frequently prescribed agents in traditional herbal medicine, and is also employed as a natural sweetening additive. In traditional Chinese medicine, licorice root is added to a variety of herbal preparations to detoxify the effects of the other herbs in the preparation. In the present study, we explored the possibility that Glycyrrhiza uralensis licorice may alleviate DOX-induced cardiotoxicity. The hexane/ethanol extract of Glycyrrhiza uralensis (HEGU), which lacks glycyrrhizin, was prepared because glycyrrhizin intake has previously been reported to induce hypertension. In an effort to determine whether HEGU ameliorates DOX-induced cytotoxicity in H9c2 rat cardiac myoblasts, the cells were pretreated with 0-15 mg/L HEGU, then treated with doxorubicin. The pretreatment of cells with HEGU resulted in a significant mitigation of DOX-induced reductions in cell numbers (34 +/- 7%) and increases in apoptosis (53 +/- 1%). The Western blot analysis of cell lysates showed that HEGU suppressed DOX-induced increases in the levels of p53, phospho-p53 (Ser 15), and Bax. In addition, HEGU induced an increase in the levels of Bcl-xL, regardless of DOX-treatment. HEGU inhibited the DOX-induced cleavage of caspases 9, 3, and 7, as well as DOX-induced poly(ADP-ribose) polymerase cleavage. Furthermore, HEGU caused reductions in the viable cell numbers of HT-29 human colon cancer cells (IC50 = 10.7 +/- 0.3 mg/L), MDA-MB-231 human breast cancer cells (IC50 = 7.5 +/- 0.1 mg/L), and DU145 human prostate cancer cells (IC50 = 4.7 +/- 0.5 mg/L). HEGU augmented DOX-induced reductions in the viability of DU145 cells (15 +/- 1%). These results indicate that HEGU may potentially be an effective agent for the alleviation of DOX-induced cardiotoxicity. PMID:18849542

Choi, Hyun Ju; Seon, Mi Ra; Lim, Soon Sung; Kim, Jong-Sang; Chun, Hyang Sook; Park, Jung Han Yoon



Protective effects of naringenin-7- O-glucoside on doxorubicin-induced apoptosis in H9C2 cells  

Microsoft Academic Search

Doxorubicin, a widely used chemotherapeutic agent, can give rise to severe cardiotoxicity by inducing cardiomyocyte apoptosis. Dracocephalum rupestre Hance, a Chinese traditional herb, has therapeutic potential for cardiovascular diseases. Naringenin-7-O-glucoside is the main active constituent of D. rupestre and there is increasing interest in its therapeutic applications. The aim of this study was to evaluate the effects of naringenin-7-O-glucoside on

Xiuzhen Han; Dongmei Ren; Peihong Fan; Tao Shen; Hongxiang Lou



Antiapoptotic effect of calcitonin gene-related peptide on oxidative stress-induced injury in H9c2 cardiomyocytes via the RAMP1\\/CRLR complex  

Microsoft Academic Search

Calcitonin gene-related peptide (CGRP) plays an important role in the mediation of protective effects observed in situations such as ischemic preconditioning in rat hearts. In this study, we investigated in H9c2 rat cardiomyoblasts if the protective effect of CGRP could be linked to an inhibitory effect on the apoptotic pathway. We also determined the specificity of observed effects by treatment

Stéphanie Sueur; Matthieu Pesant; Luc Rochette; Jean-Louis Connat



Role of Phospholipase C-?1 in Insulin-like Growth Factor I-Induced Muscle Differentiation of H9c2 Cardiac Myoblasts  

Microsoft Academic Search

Insulin-like growth factor-I (IGF-I) regulates muscle differentiation through phosphatidylinositol 3-kinase (PI 3-kinase). Also it was recently reported that PI 3-kinase is involved in the activation of phospholipase C-?1 (PLC-?1). We investigated whether PLC-?1 therefore plays a role in IGF-I-induced muscle differentiation using H9c2 rat cardiac myoblasts as a model. IGF-I was able to activate PLC-?1 via both PI 3-kinase-dependent and

Feng Hong; Keun-ai Moon; Sam Soo Kim; Young Seol Kim; Young Kil Choi; Yun Soo Bae; Pann Ghill Suh; Sung Ho Ryu; Eui-Ju Choi; Joohun Ha; Sung Soo Kim



SIH—a novel lipophilic iron chelator—protects H9c2 cardiomyoblasts from oxidative stress-induced mitochondrial injury and cell death  

Microsoft Academic Search

Recent evidence suggests that oxidative stress is a common denominator in many aspects of cardiovascular pathogenesis. Free cellular iron plays a crucial catalytic role in the formation of highly toxic hydroxyl radicals, and thereby it may aggravate the contribution of oxidative stress to cardiovascular disease. Therefore, iron chelation may be an effective therapeutic approach, but the progress in this area

Tomáš Šim?nek; Christa Boer; R. Arthur Bouwman; Ronald Vlasblom; Amanda M. G. Versteilen; Martin Št?rba; Vladimír Geršl; Radomír Hrdina; P?emysl Po?ka; Jaap J. de Lange; Walter J. Paulus; René J. P. Musters



Isoform-specific induction of PKC-? by high glucose protects heart-derived H9c2 cells against hypoxic injury  

Microsoft Academic Search

We investigated which PKC isoforms are involved in high glucose-induced protection against hypoxic injury. Treatment for 48h with high glucose (22mM) markedly increased the expression of PKC-? in the particulate fraction (213±22.1% of the control) but had no effect on other types of PKC isoforms, suggesting that the high glucose-induced increase in PKC expression is isoform-specific. The mRNA level for

Min Hwa Kim; Yi-Sook Jung; Chang-Hyun Moon; Euy-Myoung Jeong; Soo Hwan Lee; Eun Joo Baik; Chang-Kiu Moon



Effect of the Multitargeted Tyrosine Kinase Inhibitors Imatinib, Dasatinib, Sunitinib, and Sorafenib on Mitochondrial Function in Isolated Rat Heart Mitochondria and H9c2 Cells  

Microsoft Academic Search

Cardiovascular disease has recently been suggested to be a significant complication of cancer treatment with several kinase inhibitors. In some cases, the mechanisms leading to cardiotox- icity are postulated to include mitochondrial dysfunction, either as a primary or secondary effect. Detecting direct effects on mitochondrial function, such as uncoupling of oxidative phos- phorylation or inhibition of electron transport chain components,

Yvonne Will; James A. Dykens; Sashi Nadanaciva; Brad Hirakawa; Joseph Jamieson; Lisa D. Marroquin; James Hynes; Shem Patyna; Bart A. Jessen



Induction of endogenous antioxidants and phase 2 enzymes by ?-lipoic acid in rat cardiac H9C2 cells: protection against oxidative injury  

Microsoft Academic Search

?-Lipoic acid (LA) has recently been reported to exert protective effects on various forms of oxidative cardiac disorders. However, the mechanisms underlying LA-mediated cardioprotection remain to be investigated. This study was undertaken to determine whether LA treatment could increase endogenous antioxidants and phase 2 enzymes in cultured cardiomyocytes, and whether such increased cellular defenses could afford protection against oxidative cardiac

Zhuoxiao Cao; Maggie Tsang; Hai Zhao; Yunbo Li



Plantainoside D protects adriamycin-induced apoptosis in H9c2 cardiac muscle cells via the inhibition of ROS generation and NF-?B activation  

Microsoft Academic Search

Plantainoside D (PD), was isolated from the leaves of Picrorhiza scrophulariiflora (Scrophulariaceae). The anti-oxidative activity of PD was evaluated based on scavenging effects on hydroxyl radicals and superoxide anion radicals. Adriamycin (ADR) is a potent anti-tumor drug known to cause severe cardiotoxicity. Although ADR generates free radicals, the role of free radicals in the development of cardiac toxicity has not

Do-Sung Kim; Eun-Rhan Woo; Soo-Wan Chae; Ki-Chan Ha; Geum-Hwa Lee; Seong-Tshool Hong; Dae-Young Kwon; Myung-Sunny Kim; Yong-Keun Jung; Hyung-Min Kim; Hye-Kyung Kim; Hyung-Ryong Kim; Han-Jung Chae



Carvedilol inhibits mitochondrial complex I and induces resistance to H 2O 2-mediated oxidative insult in H9C2 myocardial cells  

Microsoft Academic Search

Carvedilol, a ?-adrenoreceptor antagonist with strong antioxidant activity, produces a high degree of cardioprotection in a variety of experimental models of ischemic cardiac injury. Although growing evidences suggest specific effects on mitochondrial metabolism, how carvedilol would exert its overall activity has not been completely disclosed. In the present work we have investigated the impact of carvedilol-treatment on mitochondrial bioenergetic functions

Paola Sgobbo; Consiglia Pacelli; Ignazio Grattagliano; Gaetano Villani; Tiziana Cocco



A major ozonation product of cholesterol, 3?-hydroxy-5-oxo-5,6-secocholestan-6-al, induces apoptosis in H9c2 cardiomyoblasts  

Microsoft Academic Search

Cholesterol, a major neutral lipid component of biological membranes and the lung epithelial lining fluids, is susceptible to oxidation by reactive oxygen and nitrogen species including ozone. The oxidation by ozone in biological environments results in the formation of 3?-hydroxy-5-oxo-5,6-secocholestan-6-al (cholesterol secoaldehyde or CSeco, major product) along with some other minor products. Recently, CSeco has been implicated in the pathogenesis

K. Sathishkumar; Masudul Haque; Thirugnanam E. Perumal; Joseph Francis; Rao M. Uppu



l -carnosine and verapamil inhibit hypoxia-induced expression of hypoxia inducible factor (HIF-1 ? ) in H9c2 cardiomyoblasts  

Microsoft Academic Search

Contractile failure of myocardial cells is a common cause of mortality in ischemic heart disease. In response to hypoxic conditions, cells upregulate the activity of hypoxia-inducible factor 1 (HIF-1) and express a number of genes encoding proteins that either enhance O 2delivery or increase cellular ATP levels. HIF-1 is a heterodimer of bHLH-PAS proteins, HIF-1 ?and HIF-1 ?. Both subunits

Lalita A. Bharadwaj; Gerald F. Davies; Ilungo J. Xavier; Nick Ovsenek



Peroxynitrite activates ERK via Raf1 and MEK, independently from EGF receptor and p21 Ras in H9C2 cardiomyocytes  

Microsoft Academic Search

Peroxynitrite is a potent oxidant and nitrating species proposed as a direct effector of myocardial damage in a wide range of cardiac diseases. Whether peroxynitrite also acts indirectly, by modulating cell signal transduction pathways in the myocardium, has not been investigated. Here, we examined the ability of peroxynitrite to activate extracellular signal-related kinase (ERK), a MAP kinase which has been

B. Pesse; S. Levrand; F. Feihl; B. Waeber; B. Gavillet; P. Pacher; L. Liaudet



Cell lines  

US Patent & Trademark Office Database

Genomic instability in T-antigen expressing cells can be overcome by modifying the gene expressing T-antigen so that it lacks Bub1 binding. Stable cell lines can be produced by incorporation of the modified T-antigen gene, preferably together with the catalytic sub-unit of the telomerase construct.



Characterization of Cell-Matrix Adhesion Requirements for the Formation of Fascin Microspikes  

Microsoft Academic Search

Cell adhesion to thrombospondin-1 (TSP-1) correlates with assembly of cell-substratum contact structures that contain fascin microspikes. In this analysis, cell-matrix require- ments for assembly of fascin microspikes were examined in detail. In six cell lines, cell spreading on a TSP-1 substratum correlated with expression of fascin protein and formation of fascin microspikes. Microspikes were not formed by H9c2 cells adherent

Josephine C. Adams



Expression of HSP72 after ELF-EMF exposure in three cell lines.  


It has been reported that magnetic fields with flux densities ranging from microT to mT are able to induce heat shock factor, HSP72 mRNA or heat shock proteins in various cells. In this study we investigated changes in the HSP72 mRNA transcription level in three cell lines (HL-60, H9c2, and Girardi heart cells) and in the intracellular HSP72 protein content in two cell lines (HL-60 and Girardi heart cells) after treatment schemes using electromagnetic fields with a flux density of 2 microT to 4 mT, a frequency of 50 Hz and exposure times from 15 to 30 min. None of the treatments or modalities showed any significant effect on the HSP72 protein level, although HSP72 mRNA could be induced, at least to some extent, with one of the parameter combinations in all cell lines tested. Obviously, HSP72 mRNA transcription and translation are not necessarily coupled in certain cells. This leads to the conclusion that electromagnetic field effects on HSP72 mRNA levels are not indicative for downstream effects unless increased mRNA levels can be correlated with increased HSP72 protein levels as well. PMID:17508393

Gottwald, Eric; Sontag, Werner; Lahni, Brigitte; Weibezahn, Karl-Friedrich



Mouse cell line authentication.  


The scientific community has responded to the misidentification of human cell lines with validated methods to authenticate these cells; however, few assays are available for nonhuman cell line identification. We have developed a multiplex polymerase chain reaction assay that targets nine tetranucleotide short tandem repeat (STR) markers in the mouse genome. Unique profiles were obtained from seventy-two mouse samples that were used to determine the allele distribution for each STR marker. Correlations between allele fragment length and repeat number were determined with DNA Sanger sequencing. Genotypes for L929 and NIH3T3 cell lines were shown to be stable with increasing passage numbers as there were no significant differences in fragment length with samples of low passage when compared to high passage samples. In order to detect cell line contaminants, primers for two human STR markers were incorporated into the multiplex assay to facilitate detection of human and African green monkey DNA. This multiplex assay is the first of its kind to provide a unique STR profile for each individual mouse sample and can be used to authenticate mouse cell lines. PMID:23430347

Almeida, Jamie L; Hill, Carolyn R; Cole, Kenneth D



A Correlation between Cytotoxicity and Reductase-Mediated Metabolism in Cell Lines Treated with Doxorubicin and Daunorubicin.  


The role of metabolism in daunorubicin (DAUN)- and doxorubicin (DOX)-associated toxicity in cancer patients is dependent on whether the parent drugs or major metabolites, doxorubicinol (DOXol) and daunorubicinol (DAUNol), are the more toxic species. Therefore, we examined whether an association exists between cytotoxicity and the metabolism of these drugs in cell lines from nine different tissues. Cytotoxicity studies using MTT [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide] cell viability assays revealed that four cell lines [HepG2 (liver), HCT-15 (colon), NCI-H460 (lung), and A-498 (kidney)] were more tolerant to DAUN and DOX than the five remaining cell lines [H9c2 (heart), PC-3 (prostate), OVCAR-4 (ovary), PANC-1 (pancreas), and MCF-7 (breast)], based on significantly higher LC50 values at incubation times of 6, 24, and 48 hours. Each cell line was also assessed for its efficiency at metabolizing DAUN and DOX. The four drug-tolerant cell lines converted DAUN/DOX to DAUNol/DOXol more rapidly than the five drug-sensitive cell lines. We also determined whether exposure to DAUN or DOX induced an increase in metabolic activity among any of these nine different cell types. All nine cell types showed a significant increase in their ability to metabolize DAUN or DOX in response to pre-exposure to the drug. Western blot analyses demonstrated that the increased metabolic activity toward DAUN and DOX correlated with a greater abundance of eight aldo-keto and two carbonyl reductases following exposure to either drug. Overall, our findings indicate an inverse relationship between cytotoxicity and DAUN or DOX metabolism in these nine cell lines. PMID:23995598

Bains, Onkar S; Szeitz, András; Lubieniecka, Joanna M; Cragg, Gina E; Grigliatti, Thomas A; Riggs, K Wayne; Reid, Ronald E



Ovarian granulosa cell lines  

Microsoft Academic Search

The ovary is a complex endocrine gland responsible for production of sex steroids and is the source of fertilizable ova for reproduction. It also produces various growth factors, transcription factors and cytokines that assist in the complex signaling pathways of folliculogenesis. The ovary possesses two primary steroidogenic cell types. The theca cells (and to a lesser extent, the stroma) are

Jon C. Havelock; William E. Rainey; Bruce R. Carr



Adrenocortical Cell Lines  

Microsoft Academic Search

\\u000a Initially, primary cultures of adrenocortical cells have traditionally been utilized to study the mechanisms controlling adrenocortical\\u000a steroidogenesis. However, the eventual onset of senescence in culture creates a recurring need for the costly and difficult\\u000a isolation of fresh cultures, and subsequently increases the risk of contamination. For these reasons, the use of primary cultures\\u000a has been increasingly replaced by immortalized cell

Jeniel Parmar; Anita Kulharya; William Rainey


Plant cell lines in cell morphogenesis research.  


Plant organs and tissues consist of many various cell types, often in different phases of their development. Such complex structures do not allow direct studies on behavior of individual cells. In contrast, populations of in vitro-cultured plant cells represent valuable tool for studying processes on a single-cell level, including cell morphogenesis. Here we describe characteristics of well-established model tobacco and Arabidopsis cell lines and provide detailed protocol on their cultivation, characterization, and genetic transformation. PMID:24132432

Seifertová, Daniela; Klíma, Petr; Pa?ezová, Markéta; Petrášek, Jan; Zažímalová, Eva; Opatrný, Zden?k



Thyroid cell lines in research on goitrogenesis.  


Thyroid cell lines have contributed a lot to the understanding of goitrogenesis. The cell lines mostly used in thyroid research are briefly discussed, namely the rat thyroid cell lines FRTL and FRTL-5, the porcine thyroid cell lines PORTHOS and ARTHOS, The sheep thyroid cell lines OVNIS 5H and 6H, the cat thyroid cell lines PETCAT 1 to 4 and ROMCAT, and the human thyroid cell lines FTC-133 and HTh 74. Chinese hamster ovary (CHO) cells and COS-7 cells, stably transfected with TSH receptor cDNA and expressing a functional TSH receptor, are discussed as examples for non-thyroidal cells, transfected with thyroid genes. PMID:1726925

Gerber, H; Peter, H J; Asmis, L; Studer, H



Induction of caspase-independent apoptosis in H9c2 cardiomyocytes by adriamycin treatment  

Microsoft Academic Search

The cardiotoxicity of adriamycin limits its clinical use as a powerful drug for solid tumors and malignant hematological disease. Although the precise mechanism by which it causes cardiac damage is not yet known, it has been suggested that apoptosis is the principal process in adriamycin-induced cardiomyopathy, which involves DNA fragmentation, cytochrome C release, and caspase activation. However, there has been

Ho-Joong Youn; Ho-Shik Kim; Mi-Hee Jeon; Jung-Hee Lee; Yun-Jee Seo; Yong-Joon Lee; Jeong-Hwa Lee



Human Adrenocortical Carcinoma Cell Lines  

PubMed Central

Summary The human adrenal cortex secretes mineralocorticoids, glucocorticoids and adrenal androgens. These steroids are produced from unique cell types located within the three distinct zones of the adrenal cortex. Disruption of adrenal steroid production results in a variety of diseases that can lead to hypertension, metabolic syndrome, infertility and androgen excess. The adrenal cortex is also a common site for the development of adenomas, and rarely the site for the development of carcinomas. The adenomas can lead to diseases associated with adrenal steroid excess, while the carcinomas are particularly aggressive and have a poor prognosis. In vitro cell culture models provide an important tool to examine molecular and cellular mechanisms controlling both the normal and pathologic function of the adrenal cortex. Herein we discuss the human adrenocortical cell lines and their use as model systems for adrenal studies.

Wang, Tao; Rainey, William E.



Transmission line analysis of MRAM cell  

Microsoft Academic Search

A test configuration of magnetic random access memory (MRAM) unit cell was modeled and its three-dimensional finite element method (FEM) analysis was utilized to calculate S-parameters. Transmission line analysis was performed for word line, bit line and readout signal path. Line width was varied from 0.5 to 2 ?m, line thickness from 0.1 to 1 ?m, and line length from

S. Jo



Development and characterization of insect cell lines  

Microsoft Academic Search

With the wide availability of insect cell culture media, it can generally be considered a routine process to develop new cell lines. Exceptions to this statement do exist, of course. Difficulties may arise when attempting to culture a specific cell type. For example, while there are a few cell lines from insect fat body and at least one from the

Dwight E. Lynn



Molecular Characterization of Putative Chordoma Cell Lines  

PubMed Central

Immortal tumor cell lines are an important model system for cancer research, however, misidentification and cross-contamination of cell lines are a common problem. Seven chordoma cell lines are reported in the literature, but none has been characterized in detail. We analyzed gene expression patterns and genomic copy number variations in five putative chordoma cell lines (U-CH1, CCL3, CCL4, GB60, and CM319). We also created a new chordoma cell line, U-CH2, and provided genotypes for cell lines for identity confirmation. Our analyses revealed that CCL3, CCL4, and GB60 are not chordoma cell lines, and that CM319 is a cancer cell line possibly derived from chordoma, but lacking expression of key chordoma biomarkers. U-CH1 and U-CH2 both have gene expression profiles, copy number aberrations, and morphology consistent with chordoma tumors. These cell lines also harbor genetic changes, such as loss of p16, MTAP, or PTEN, that make them potentially useful models for studying mechanisms of chordoma pathogenesis and for evaluating targeted therapies.

Bruderlein, Silke; Sommer, Joshua B.; Meltzer, Paul S.; Li, Sufeng; Osada, Takuya; Ng, David; Moller, Peter; Alcorta, David A.; Kelley, Michael J.



CellLineMiner: a knowledge portal for human cell lines  

PubMed Central

Experimental models of human tissues and disease phenotypes frequently rely upon immortalized cell lines, which are easily accessible and simple to use due to their infinite capability of cell division. For decades, cell lines have been used to investigate cellular mechanisms of disease and the efficacy of drugs, most prominently for human cancers. However, the large body of knowledge with respect to human cell lines exists primarily in an unstructured fashion, that is, as free text in the scientific literature. Here we present CellLineMiner, a novel text mining-based web database that provides a comprehensive view of human cell line knowledge. The application offers a simple search in all indexed cell lines, accompanied by a rapid display of all identified literature associations. The CellLineMiner is intended to serve as a knowledge resource companion to the cellular model systems used in biomedical research. Availability CellLineMiner is accessible at

Nakken, Sigve; Johansen, Morten; Fillebeen, Julien; Berge, Ole Petter; Kirker?d, Harald; Jenssen, Tor-Kristian; Hovig, Eivind



Cell culturing of human and murine microglia cell lines.  


Despite the fact that microglia cells were first described almost a century ago, microglia-derived immortalized cell lines have only been established in the last two decades. One should be aware of their limitations but also of their advantages. Cell lines offer a potentially powerful tool to investigate some functional aspects of microglia. Cell culturing of human and murine microglia cell lines will be described in this chapter. It includes a presentation of equipment needed, cell culture medium and supplements, cell culture monitoring, and a protocol describing the steps for subculturing of microglia cell lines. PMID:23813364

Rodhe, Johanna



Red blood cell production from immortalized progenitor cell line  

Microsoft Academic Search

The supply of transfusable red blood cells (RBCs) is not sufficient in many countries. If immortalized erythroid progenitor\\u000a cell lines able to produce transfusable RBCs in vitro were established, they would be valuable resources. However, such cell\\u000a lines have not been established. We have developed a robust method to establish immortalized erythroid progenitor cell lines\\u000a following the induction of hematopoietic

Yukio Nakamura; Takashi Hiroyama; Kenichi Miharada; Ryo Kurita



Cell Lines Secreting Uteroglobin In vitro.  

National Technical Information Service (NTIS)

The invention is two immortal cell lines that secrete uteroglobin in vitro when stimulated by a steroid hormone. The invention includes a method for screening the steroid stimulating property of a compound.

A. B. Mukherjee J. Y. Chou



Ecdysone action on insect cell lines  

Microsoft Academic Search

Summary  Cell lines derived from embryos of the tobacco hornworm,Manduca sexta (L.), showed a marked morphological response to treatment with physiological doses of ?-ecdysone. The response of these cell\\u000a lines with ?-ecdysone indicated that the penta-ol (?-ecdysone) must be converted to the hexa-ol (?-ecdysone) form before the\\u000a morphological response can appear. Liquid chromatographic analysis of the spent medium confirmed that the

Edwin P. Marks; G. Mark Holman



Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines  

PubMed Central

The Cell Line Data Base (CLDB) is a well-known reference information source on human and animal cell lines including information on more than 6000 cell lines. Main biological features are coded according to controlled vocabularies derived from international lists and taxonomies. HyperCLDB ( is a hypertext version of CLDB that improves data accessibility by also allowing information retrieval through web spiders. Access to HyperCLDB is provided through indexes of biological characteristics and navigation in the hypertext is granted by many internal links. HyperCLDB also includes links to external resources. Recently, an interest was raised for a reference nomenclature for cell lines and CLDB was seen as an authoritative system. Furthermore, to overcome the cell line misidentification problem, molecular authentication methods, such as fingerprinting, single-locus short tandem repeat (STR) profile and single nucleotide polymorphisms validation, were proposed. Since this data is distributed, a reference portal on authentication of human cell lines is needed. We present here the architecture and contents of CLDB, its recent enhancements and perspectives. We also present a new related database, the Cell Line Integrated Molecular Authentication (CLIMA) database (, that allows to link authentication data to actual cell lines.

Romano, Paolo; Manniello, Assunta; Aresu, Ottavia; Armento, Massimiliano; Cesaro, Michela; Parodi, Barbara



Fish cell lines: Establishment and characterization of nine cell lines from salmonids  

Microsoft Academic Search

Summary  Nine permanent cell lines have been established from five species of salmonids native to America's Pacific Northwest. With\\u000a the exception of a hepatoma from an adult trout, the lines were derived from normal tissues of embryonic or juvenile fish.\\u000a Cells were routinely grown in Eagle's minimum essential medium with 10% fetal bovine serum. Optimum growth temperatures for\\u000a these lines ranged

C. N. Lannan; J. R. Winton; J. L. Fryer



Anomalous dystroglycan in carcinoma cell lines  

Microsoft Academic Search

Dystroglycan is a receptor responsible for crucial interactions between extracellular matrix and cytoplasmic space. We provide the first evidence that dystroglycan is truncated. In HC11 normal murine and the 184B5 non-tumorigenic mammary human cell lines, the expected ?-dystroglycan 43 kDa band was found but human breast T47D, BT549, MCF7, colon HT29, HCT116, SW620, prostate DU145 and cervical HeLa cancer cells

Carmen Losasso; Francesca Di Tommaso; Alessandro Sgambato; Raffaele Ardito; Achille Cittadini; Bruno Giardina; Tamara C. Petrucci; Andrea Brancaccio



Peptidomic analysis of human cell lines  

PubMed Central

Peptides have been proposed to function in intracellular signaling within the cytosol. Although cytosolic peptides are considered to be highly unstable, a large number of peptides have been detected in mouse brain and other biological samples. In the present study, we evaluated the peptidome of three diverse cell lines: SH-SY5Y, MCF7, and HEK293 cells. A comparison of the peptidomes revealed considerable overlap in the identity of the peptides found in each cell line. The majority of the observed peptides are not derived from the most abundant or least stable proteins in the cell, and approximately half of the cellular peptides correspond to the N- or C- termini of the precursor proteins. Cleavage site analysis revealed a preference for hydrophobic residues in the P1 position. Quantitative peptidomic analysis indicated that the levels of most cellular peptides are not altered in response to elevated intracellular calcium, suggesting that calpain is not responsible for their production. The similarity of the peptidomes of the three cell lines and the lack of correlation with the predicted cellular degradome implies the selective formation or retention of these peptides, consistent with the hypothesis that they are functional in the cells.

Gelman, Julia S.; Sironi, Juan; Castro, Leandro M.; Ferro, Emer S.; Fricker, Lloyd D.



Fish cell lines: Establishment and characterization of three new cell lines from grass carp ( Ctenopharyngodon idella )  

Microsoft Academic Search

Summary  Three new cell lines were established from tissues of the grass carp,Ctenopharyngodon idella. Derived from the fin, snout, and swim bladder of two apparently healthy diploid fry, these cell lines have been designated\\u000a GCF, GCS-2, and GCSB, respectively. The cells grew at temperatures between 24° and 36° C with optimal growth at 32° C and\\u000a have been subcultured more than

Yuanan Lu; C. N. Lannan; J. S. Rohovec; J. L. Fryer



The clinical relevance of cancer cell lines.  


Although advances in genomics during the last decade have opened new avenues for translational research and allowed the direct evaluation of clinical samples, there is still a need for reliable preclinical models to test therapeutic strategies. Human cancer-derived cell lines are the most widely used models to study the biology of cancer and to test hypotheses to improve the efficacy of cancer treatment. Since the development of the first cancer cell line, the clinical relevance of these models has been continuously questioned. Based upon recent studies that have fueled the debate, we review the major events in the development of the in vitro models and the emergence of new technologies that have revealed important issues and limitations concerning human cancer cell lines as models. All cancer cell lines do not have equal value as tumor models. Some have been successful, whereas others have failed. However, the success stories should not obscure the growing body of data that motivates us to develop new in vitro preclinical models that would substantially increase the success rate of new in vitro-assessed cancer treatments. PMID:23434901

Gillet, Jean-Pierre; Varma, Sudhir; Gottesman, Michael M



Characterization of a new megakaryocytic cell line: the Dami cell  

Microsoft Academic Search

A new human megakaryocytic cell line (Dami) has been established from the blood of a patient with megakaryo- blastic leukemia. The Dami cells grow primarily in suspen- sion with a doubling time of 24 to 30 hours. By light and electron microscopy. the Dami cells range in size from 1 2 to 120 ?tm in diameter and have lobulated nuclei

SM Greenberg; DS Rosenthal; TA Greeley; R Tantravahi; RI Handin



MicroRNA profiling during rat ventricular maturation: A role for miR-29a in regulating cardiomyocyte cell cycle re-entry.  


Recent studies demonstrated that the mammalian heart possesses some capacity to proliferate. We observed cardiomyocyte proliferation within 4 weeks of age (P4W) in rats. We found 95 microRNAs that are differentially expressed in P4W cardiomyocytes. MicroRNA-29a was among the most highly up-regulated microRNAs in P4W cardiomyocytes. Overexpression of microRNA-29a suppressed the proliferation of H9c2 cell line. MicroRNA-29a inhibition induced cardiomyocytes to proliferate, accelerated the G1/S and G2/M transition, and up-regulated the cell cycle gene expression. Cyclin D2 (CCND2) was identified as a direct target of microRNA-29a. These findings indicate that microRNA-29a is involved in cardiomyocyte proliferation during postnatal development. PMID:23587482

Cao, Xiaoqing; Wang, Jue; Wang, Zhenhua; Du, Juan; Yuan, Xin; Huang, Weicong; Meng, Jian; Gu, Haiyong; Nie, Yu; Ji, Bingyang; Hu, Shengshou; Zheng, Zhe



Glioma Cell Lines: Role of Cancer Stem Cells  

Microsoft Academic Search

\\u000a In this chapter, we review the cancer stem cell hypothesis and discuss implications for this paradigm in considering whether\\u000a glioma cell lines contain bonafide cancer stem cells, the source material used for tissue culture, and experimental methods\\u000a used in preclinical research. We identify three key modifications to standard tissue culture protocols that allow for enrichment\\u000a of cancer stem cells that

John R. Ohlfest; Stacy A. Decker


New cell lines with chondrocytic phenotypes from human chondrosarcoma  

Microsoft Academic Search

In the present study, we investigated chondrocytic characterization for newly established human chondrosarcoma cell lines. A chondrosarcoma cell line, HCS-TG, was established by the implantation of grade-2 human chondrosarcoma into athymic mice. Cloning of HCS-TG cells from passage 17 was performed. After cell cloning, two clonal-cell lines (HCS-TG C3 and E2) with good proliferative activities were obtained. These cell lines

Ikuo Kudawara; Nobuhito Araki; Akira Myoui; Yoichi Kato; Atsumasa Uchida; Hideki Yoshikawa



Peroxisome proliferator-activated receptor y (PPARy) activation protects H9c2 cardiomyoblasts from oxidative stress-induced apoptosis  

Microsoft Academic Search

Objective: Activation of peroxisome proliferator-activated receptor a (PPARa) and PPARg plays beneficial roles in cardiovascular disorders such as atherosclerosis and heart reperfusion. Although PPARa and g have been documented to reduce oxidative stress in the vasculature and the heart, the role of PPARy remains poorly studied. Methods and results: We focused on PPARy function in the regulation of oxidative stress-induced

Matthieu Pesant; Stephanie Sueur; Patrick Dutartre; Mireille Tallandier; Paul A. Grimaldi; Luc Rochette; Jean-Louis Connat



Establishment and characterisation of two novel breast cancer cell lines  

Microsoft Academic Search

Two novel oestrogen receptor (ER) negative breast cancer cell lines, BCa-11 (familial) and BCa-15 (sporadic) were successfully established from primary tumours. Characterisation of these cell lines showed expression of epithelial specific antigen and cytokeratins confirming their epithelial lineage. Analysis of ultrastructure and anchorage independent growth confirmed the epithelial nature and transformed phenotype of these cells. Both cell lines showed loss

Sarangadhara Appala Raju Bagadi; Jatinder Kaur; Ranju Ralhan



Generation of Human Pulmonary Microvascular Endothelial Cell Lines  

Microsoft Academic Search

The limited lifespan of human microvascular endothelial cells in cell culture represents a major obstacle for the study of microvascular pathobiology. To date, no endothelial cell line is available that demonstrates all of the fundamental characteristics of microvascular endothelial cells. We have generated endothelial cell lines from human pulmonary microvascular endothelial cells (HPMEC) isolated from adult donors. HPMEC were cotransfected

Vera Krump-Konvalinkova; Fernando Bittinger; Ronald E Unger; Kirsten Peters; Hans-Anton Lehr; C James Kirkpatrick



CellLineNavigator: a workbench for cancer cell line analysis  

PubMed Central

The CellLineNavigator database, freely available at, is a web-based workbench for large scale comparisons of a large collection of diverse cell lines. It aims to support experimental design in the fields of genomics, systems biology and translational biomedical research. Currently, this compendium holds genome wide expression profiles of 317 different cancer cell lines, categorized into 57 different pathological states and 28 individual tissues. To enlarge the scope of CellLineNavigator, the database was furthermore closely linked to commonly used bioinformatics databases and knowledge repositories. To ensure easy data access and search ability, a simple data and an intuitive querying interface were implemented. It allows the user to explore and filter gene expression, focusing on pathological or physiological conditions. For a more complex search, the advanced query interface may be used to query for (i) differentially expressed genes; (ii) pathological or physiological conditions; or (iii) gene names or functional attributes, such as Kyoto Encyclopaedia of Genes and Genomes pathway maps. These queries may also be combined. Finally, CellLineNavigator allows additional advanced analysis of differentially regulated genes by a direct link to the Database for Annotation, Visualization and Integrated Discovery (DAVID) Bioinformatics Resources.

Krupp, Markus; Itzel, Timo; Maass, Thorsten; Hildebrandt, Andreas; Galle, Peter R.; Teufel, Andreas



Pancreastatin producing cell line from human pancreatic islet cell tumor.  


It has been characterized that cell line QGP-1 derived from human non-functioning pancreatic islet cell tumor produces human pancreastatin. Exponentially growing cultures produced 5.7 fmol of pancreastatin/10(6) cells/hr. Human pancreastatin immunoreactivities in plasma and tumor after xenografting with QGP-1 into nude mouse were 92.7 fmol/ml and 160.2 pmol/g wet weight, respectively. Immunocytochemical study revealed both chromogranin A and pancreastatin immunoreactive cells in the tumor. Gel filtrations of culture medium and tumor extract identified heterogenous molecular forms of PST-LI which eluted as large and smaller molecular species. These results suggest that plasma pancreastatin levels may be useful as a tumor marker of endocrine tumor of the pancreas, and the pancreastatin producing cell line may be useful for studies of the mechanism of secretions and processing of chromogranin A and pancreastatin. PMID:2159299

Funakoshi, A; Tateishi, K; Tsuru, M; Jimi, A; Wakasugi, H; Ikeda, Y; Kono, A



A bioinformatics analysis of the cell line nomenclature  

PubMed Central

Motivation: Cell lines are used extensively in biomedical research, but the nomenclature describing cell lines has not been standardized. The problems are both linguistic and experimental. Many ambiguous cell line names appear in the published literature. Users of the same cell line may refer to it in different ways, and cell lines may mutate or become contaminated without the knowledge of the user. As a first step towards rationalizing this nomenclature, we created a cell line knowledgebase (CLKB) with a well-structured collection of names and descriptive data for cell lines cultured in vitro. The objectives of this work are: (i) to assist users in extracting useful information from biomedical text and (ii) to highlight the importance of standardizing cell line names in biomedical research. This CLKB contains a broad collection of cell line names compiled from ATCC, Hyper CLDB and MeSH. In addition to names, the knowledgebase specifies relationships between cell lines. We analyze the use of cell line names in biomedical text. Issues include ambiguous names, polymorphisms in the use of names and the fact that some cell line names are also common English words. Linguistic patterns associated with the occurrence of cell line names are analyzed. Applying these patterns to find additional cell line names in the literature identifies only a small number of additional names. Annotation of microarray gene expression studies is used as a test case. The CLKB facilitates data exploration and comparison of different cell lines in support of clinical and experimental research. Availability: The web ontology file for this cell line collection can be downloaded at Contact: Supplementary information: Supplementary data are available at Bioinformatics online.

Sarntivijai, Sirarat; Ade, Alexander S.; Athey, Brian D.; States, David J.



On the Ontology Based Representation of Cell Lines  

PubMed Central

Cell lines are frequently used as highly standardized and reproducible in vitro models for biomedical analyses and assays. Cell lines are distributed by cell banks that operate databases describing their products. However, the description of the cell lines' properties are not standardized across different cell banks. Existing cell line-related ontologies mostly focus on the description of the cell lines' names, but do not cover aspects like the origin or optimal growth conditions. The objective of this work is to develop an ontology that allows for a more comprehensive description of cell lines and their metadata, which should cover the data elements provided by cell banks. This will provide the basis for the standardized annotation of cell lines and corresponding assays in biomedical research. In addition, the ontology will be the foundation for automated evaluation of such assays and their respective protocols in the future. To accomplish this, a broad range of cell bank databases as well as existing ontologies were analyzed in a comprehensive manner. We identified existing ontologies capable of covering different aspects of the cell line domain. However, not all data fields derived from the cell banks' databases could be mapped to existing ontologies. As a result, we created a new ontology called cell culture ontology (CCONT) integrating existing ontologies where possible. CCONT provides classes from the areas of cell line identification, origin, cell line properties, propagation and tests performed.

Ganzinger, Matthias; He, Shan; Breuhahn, Kai; Knaup, Petra



Establishment and characterization of four human pancreatic carcinoma cell lines  

Microsoft Academic Search

We characterized four pancreatic carcinoma cell lines (designated SNU-213, SNU-324, SNU-410, and SNU-494) established from histopathologically varied primary or liver metastatic tumor samples of Korean patients. Three cell lines grew as adherent monolayers and one as adherent and floating cell clumps. All lines had: (1) relatively high viability; (2) an absence of mycoplasma or bacterial contamination; (3) genetic heterogeneity as

Ja-Lok Ku; Kyong-Ah Yoon; Woo-Ho Kim; Jin Jang; Kyung-Suk Suh; Sun-Whe Kim; Yong-Hyun Park; Jae-Gahb Park



Human Myeloma Cell Line Carrying a Philadelphia Chromosome  

Microsoft Academic Search

A new human plasmacytoma cell line (Karpas 707) has been established from a myeloma patient. The cultured cells are negative for Epstein-Barr viral nuclear antigen and free of mycoplasma. They are similar to plasma cells and secrete only lambda light chains. The cells are hypodiploid and contain the Philadelphia chromosome and other abnormalities. This cell line may be suitable for

Abraham Karpas; Patricia Fischer; David Swirsky



Role of Glial Cell Line-Derived Neurotrophic Factor in Germ-Line Stem Cell Fate  

PubMed Central

The overall goal of this study is to unravel the role(s) played by glial cell line-derived neurotrophic factor (GDNF) in the fate of spermatogonial stem cells. There is great interest in the biology of spermatogonial stem cells, or Asingle spermatogonia, because of their importance in the treatment of infertility, the development of contraceptives, and the understanding of the etiology of testicular cancer, particularly seminoma. In the mouse, spermatogonial stem cells express GFR?-1, the receptor for GDNF, and respond to this growth factor in vivo and in vitro. GDNF is produced by the adjacent Sertoli cells, which are part of the germ-line stem cell niche in vertebrates. We specifically isolated GFR?-1–positive spermatogonia using an immunomagnetic bead technique. We then stimulated the cells with 100 ng/mL of rGDNF for 10 hours; unstimulated cells served as negative controls. Microarray analysis, immunocytochemistry, and Western blotting revealed that Numb, a regulator of the Notch pathway, is upregulated by GDNF in spermatogonial stem cells. There are indications that in rats, mice, and humans, the Notch pathway promotes spermatogonial differentiation. We observed that an increase in Numb expression is concomitant with Notch degradation in these cells. Thus, through Numb, GDNF might inhibit differentiation and allows the maintenance of the stem cell pool in the mouse seminiferous epithelium.

Braydich-Stolle, Laura; Nolan, Courtney; Dym, Martin; Hofmann, Marie-Claude



Tetanus toxin as a marker for small-cell lung cancer cell lines  

Microsoft Academic Search

Tetanus toxin labeling of human lung cancer cell lines was investigated using direct and indirect immunofluorescence and immunohistochemical staining. Cells of characterized permanent cell lines, eight small-cell lung cancer (SCLC) cell lines of classic subtype, six SCLC cell lines of variant subtype and seven non-small-cell lung cancer (NSCLC) cell lines, were incubated with a saturating concentration of tetanus toxin. For

Jochen Heymanns; Kurt Neumann; Klaus Havemann



Detection Algorithm for the Validation of Human Cell Lines  

PubMed Central

Cell lines are an important tool in understanding all aspects of cancer growth, development, metastasis, and tumor cell death. There has been a dramatic increase in the number of cell lines and diversity of the cancers they represent; however, misidentification and cross-contamination of cell lines can lead to erroneous conclusions. One method that has gained favor for authenticating cell lines is the use of short tandem repeats (STR) to generate a unique DNA profile. The challenge in validating cell lines is the requirement to compare the large number of existing STR profiles against cell lines of interest, particularly when considering that the profiles of many cell lines have drifted over time and original samples are not available. We report here methods that analyze the variations and the proportional changes extracted from tetra-nucleotide repeat regions in the STR analysis. This technique allows a paired match between a target cell line and a reference database of cell lines to find cell lines that match within a user designated percentage cut-off quality matrix. Our method accounts for DNA instability and can suggest whether the target cell lines are misidentified or unstable.

Eltonsy, Nevine; Gabisi, Vivian; Li, Xuesong; Russe, K. Blair; Mills, Gordon B.; Stemke-Hale, Katherine



Road for understanding cancer stem cells: model cell lines.  


There is increasing evidence suggesting that stem cells are susceptive to carcinogenesis and, consequently, can be the origin of many cancers. Recently, the neoplastic potential of stem cells has been supported by many groups showing the existence of subpopulations with stem cell characteristics in tumor biopsies such as brain and breast. Evidence supporting the cancer stem cell hypothesis has gained impact due to progress in stem cell biology and development of new models to validate the self-renewal potential of stem cells. Recent evidence on the possible identification of cancer stem cells may offer an opportunity to use these cells as future therapeutic targets. Therefore, model systems in this field have become very important and useful. This review will focus on the state of knowledge on cancer stem cell research, including cell line models for cancer stem cells. The latter will, as models, help us both in the identification and characterization of cancer stem cells and in the further development of therapeutic strategies including tissue engineering. PMID:18034633

Serakinci, Nedime; Erzik, Can



Match criteria for human cell line authentication: where do we draw the line?  


Continuous human cell lines have been used extensively as models for biomedical research. In working with these cell lines, researchers are often unaware of the risk of cross-contamination and other causes of misidentification. To reduce this risk, there is a pressing need to authenticate cell lines, comparing the sample handled in the laboratory to a previously tested sample. The American Type Culture Collection Standards Development Organization Workgroup ASN-0002 has developed a Standard for human cell line authentication, recommending short tandem repeat (STR) profiling for authentication of human cell lines. However, there are known limitations to the technique when applied to cultured samples, including possible genetic drift with passage. In our study, a dataset of 2,279 STR profiles from four cell banks was used to assess the effectiveness of the match criteria recommended within the Standard. Of these 2,279 STR profiles, 1,157 were grouped into sets of related cell lines-duplicate holdings, legitimately related samples or misidentified cell lines. Eight core STR loci plus amelogenin were used to unequivocally authenticate 98% of these related sets. Two simple match algorithms each clearly discriminated between related and unrelated samples, with separation between related samples at ?80% match and unrelated samples at <50% match. A small degree of overlap was noted at 50-79% match, mostly from cell lines known to display variable STR profiles. These match criteria are recommended as a simple and effective way to interpret results from STR profiling of human cell lines. PMID:23136038

Capes-Davis, Amanda; Reid, Yvonne A; Kline, Margaret C; Storts, Douglas R; Strauss, Ethan; Dirks, Wilhelm G; Drexler, Hans G; MacLeod, Roderick A F; Sykes, Gregory; Kohara, Arihiro; Nakamura, Yukio; Elmore, Eugene; Nims, Raymond W; Alston-Roberts, Christine; Barallon, Rita; Los, Georgyi V; Nardone, Roland M; Price, Paul J; Steuer, Anton; Thomson, Jim; Masters, John R W; Kerrigan, Liz



Distinct differentiation characteristics of individual human embryonic stem cell lines  

Microsoft Academic Search

BACKGROUND: Individual differences between human embryonic stem cell (hESC) lines are poorly understood. Here, we describe the derivation of five hESC lines (called FES 21, 22, 29, 30 and 61) from frozen-thawed human embryos and compare their individual differentiation characteristic. RESULTS: The cell lines were cultured either on human or mouse feeder cells. The cells grew significantly faster and could

Milla Mikkola; Cia Olsson; Jaan Palgi; Jarkko Ustinov; Tiina Palomaki; Nina Horelli-Kuitunen; Sakari Knuutila; Karolina Lundin; Timo Otonkoski; Timo Tuuri



Human embryonic stem cell lines derived from the Chinese population  

Microsoft Academic Search

Six human embryonic stem cell lines were established from surplus blastocysts. The cell lines expressed alkaline phosphatase and molecules typical of primate embryonic stem cells, including Oct-4, Nanog, TDGF1, Sox2, EBAF, Thy-1, FGF4, Rex-1, SSEA-3, SSEA-4, TRA-1-60 and TRA-1–81. Five of the six lines formed embryoid bodies that expressed markers of a variety of cell types; four of them formed

Zhen Fu FANG; Fan JIN; Hui GAI; Ying CHEN; Li WU; Ai Lian LIU; Bin CHEN; Hui Zhen SHENG



Establishment and characterization of human neuroblastoma cell lines.  


Three new tissue culture cell lines, CHP-100, CHP-126, and CHP-134, have been established from explant cultures of human neuroblastoma. The cell lines have been characterized with respect to morphology, chromosomes constitution, growth, neural enzyme content, and their ability to grow in nude mice. The cells grow as dense masses comprised of fibroblast-or neuroblast-like cells with small processes. The cell lines differ in their neural enzyme acitivity. The chromosomal content of the 3 cell lines is near diploid, and all are capable of forming tumors in nude mice. The morphological findings indicate that the cells in culture resemble those found in the tumor, and the enzyme activities are consistent with those of nervous tissue. This the morphological, biochemical, and tumorigenic properties confirm that the 3 cell lines are neoplastic cells of neural origin. PMID:10079

Schlesinger, H R; Gerson, J M; Moorhead, P S; Maguire, H; Hummeler, K



Cell line misidentification: the beginning of the end.  


Cell lines are used extensively in research and drug development as models of normal and cancer tissues. However, a substantial proportion of cell lines is mislabelled or replaced by cells derived from a different individual, tissue or species. The scientific community has failed to tackle this problem and consequently thousands of misleading and potentially erroneous papers have been published using cell lines that are incorrectly identified. Recent efforts to develop a standard for the authentication of human cell lines using short tandem repeat profiling is an important step to eradicate this problem. PMID:20448633



Characterization of a transformed rat retinal ganglion cell line  

Microsoft Academic Search

The purpose of the present study was to establish a rat retinal ganglion cell line by transformation of rat retinal cells. For this investigation, retinal cells were isolated from postnatal day 1 (PN1) rats and transformed with the ?2 E1A virus. In order to isolate retinal ganglion cells (RGC), single cell clones were chosen at random from the transformed cells.

R. R. Krishnamoorthy; P. Agarwal; G. Prasanna; K. Vopat; W. Lambert; H. J. Sheedlo; I.-H. Pang; D. Shade; R. J. Wordinger; T. Yorio; A. F Clark; N. Agarwal



Growth properties and alloantigenic expression of murine lymphoblastoid cell lines  

PubMed Central

Murine lymphoblastoid cell lines were evaluated for their expression of Thy-1 and thymus leukemia (TL) differentiation alloantigens. Two culture conditions were shown to affect this expression. Cells grown in fetal bovine serum (FBS)-enriched medium expressed up to 15 times the amount of TL as cells grown in horse serum (HS)-enriched medium. Thy-1 expression was less affected by the type of serum used for culture. The phase of growth when the cells were harvested, was demonstrated to affect the expression of Thy-1. The expression of Thy- 1.2 for one cell line examined, L-251A, during logarithmic growth was threefold greater than cells collected during either lag or stationary growth. When culture conditions were standardized a ranking of the amount of Thy-1 and TL expressed by several cell lines was made. All cell lines, except one, L-1210, expressed Thy-1. There was a 450-fold difference in the expression of Thy-1 between the cell lines evaluated. Seven cell lines expressed TL-1,2,3 with a ninefold difference in the amount of expression. The L-251A cell line was cultured in a 14 liter fermentor for a 26 day period. During this time TL and Thy-1 expression did not vary significantly, demonstrating that lymphoblastoid cell lines can be cultured on a continuous basis and will continue to express their surface alloantigens.



Human embryonic stem cell lines with genetic disorders  

Microsoft Academic Search

A previous study described the establishment of human embryonic stem cell (ESC) lines from different sources of embryonic material, including morula, whole blastocyst and isolated inner cell mass. Using these methods, a repository of ESC lines has been established with different genetic abnormalities, which provides an unlimited source of disease cells in culture for undertaking research on the primary disturbances

Y Verlinsky; N Strelchenko; V Kukharenko; S Rechitsky; O Verlinsky; V Galat; A Kuliev



Characterization and Analysis of Human Chordoma Cell Lines  

PubMed Central

Study Design An experimental study to investigate the characterization of 3 chordoma cell lines. Objective To characterize chordoma cell lines and generate hypothesis for further chordoma studies. Summary of Background Data Three cultured human chordoma cell lines have been successfully generated; however, their characterization is incomplete. Complete characterization of chordoma cell lines is necessary for these reagents to be a useful preclinical model. Methods Three chordoma cell lines, CH 8, U-CH1, and GP 60, were cultured in different commercially available tissue culture media. They were also cultured in different environments, which included collagen substrate, various concentrations of glucose, and various levels of hypoxic conditions. The rate of cell proliferation was assessed by either MTT or numeration assay. A 3-dimensional (3D) cell culture model of these chordoma cell lines was also studied, and the expression of vimentin and cytokeratin was measured by immunofluorescence and Western blot. Additionally, the sensitivity of the 3 chordoma cell lines to 6 chemotherapeutic drugs was analyzed. Results CH 8, GP 60, and U-CH1 cells proliferate more actively in Iscove Modified Dulbecco Medium or Dulbecco modified Eagle Medium and less actively in RPMI medium. All 3 chordoma cell lines universally grow better in collagen substrate and survive in hypoxic conditions, whereas glucose concentration has no significant influence on their growth properties. Chordoma cell lines grew well in 3D culture systems and formed acini-like spheroids and retained the expression of vimentin and cytokeratin. MTT analysis indicates that all 3 chordoma cell lines are sensitive to doxorubicin, yondelis, zalypsis, and cisplatin. Conclusion We characterized 3 chordoma cell lines for differential growth properties in a variety of media and response to chemotherapeutic agents.

Yang, Cao; Hornicek, Francis J.; Wood, Kirkham B.; Schwab, Joseph H.; Choy, Edwin; Iafrate, John; Rosenberg, Andrew; Nielsen, G. Petur; Xavier, Ramnik J.; Mankin, Henry; Duan, Zhenfeng



Antiproliferative action of metformin in human lung cancer cell lines.  


The oral antidiabetic agent metformin has anticancer properties, probably via adenosine monophosphate-activated protein kinase activation. In the present study, growth inhibition was assessed by a clonogenic and by a cell survival assay, apoptosis induction was assessed by Hoechst staining and caspase activities and cell cycle alteration after exposure to metformin, and the interaction of metformin with cisplatin in vitro were elucidated in four human lung cancer cell lines representing squamous, adeno-, large cell and small cell carcinoma. Clonogenicity and cell proliferation were inhibited by metformin in all the cell lines examined. This inhibitory effect was not specific to cancer cells because it was also observed in a non-transformed human mesothelial cell line and in mouse fibroblast cell lines. Inhibition of clonogenicity was observed only when the cells were exposed to metformin for a long period, (10 days) and the surviving fraction, obtained after inhibiting proliferation by increasing the dose, reached a plateau at approximately 0.1-0.3, indicating the cytostatic characteristics of metformin. Metformin induced significant apoptosis only in the small cell carcinoma cell line. A tendency of cell cycle accumulation at the G0/G1 phase was observed in all four cell lines. Cisplatin, in a dose-dependent manner, severely antagonized the growth inhibitory effect of metformin, and even reversed the effect in three cell lines but not in the adenocarcinoma cell line. The present data obtained using various histological types of human lung cancer cell lines in vitro illustrate the cytostatic nature of metformin and its cytoprotective properties against cisplatin. PMID:22576795

Ashinuma, Hironori; Takiguchi, Yuichi; Kitazono, Satoru; Kitazono-Saitoh, Miyako; Kitamura, Atsushi; Chiba, Tetsuhiro; Tada, Yuji; Kurosu, Katsushi; Sakaida, Emiko; Sekine, Ikuo; Tanabe, Nobuhiro; Iwama, Atsushi; Yokosuka, Osamu; Tatsumi, Koichiro



Hyaluronan production regulation from porcine hyalocyte cell line by cytokines  

Microsoft Academic Search

The objective of this study were to establish a cell line derived from porcine hyalocytes and to investigate the regulation of hyaluronan (HA) synthesis in response to cytokines. After 50 passages of the cells derived from porcine vitreous tissue, a cell line was generated. The immortalized cells showed fibroblastic morphology. The cell doubling time was 56.9h. In the mRNA level,

Koichi Nishitsuka; Yoshiko Kashiwagi; Naoki Tojo; Chikako Kanno; Yoshinori Takahashi; Teiko Yamamoto; Paraskevi Heldin; Hidetoshi Yamashita



Establishment and therapeutic use of human embryonic stem cell lines  

Microsoft Academic Search

Embryonic stem (ES) cell lines, which are derived from the inner cell mass of blastocysts, proliferate indefinitely in vitro,\\u000a retaining their potency to differentiate into various cell types derived from all of the three embryonic germ layers: the\\u000a ectoderm, mesoderm and endoderm. Establishment of human ES cell lines in 1 998 has indicated the great potential of ES cells\\u000a for

Hirofumi Suemori



Clonal derivation and characterization of human embryonic stem cell lines  

Microsoft Academic Search

Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived

Nico Heins; Anders Lindahl; Ulrika Karlsson; Marie Rehnström; Gunilla Caisander; Katarina Emanuelsson; Charles Hanson; Henrik Semb; Petter Björquist; Peter Sartipy; Johan Hyllner



Comparative recombinant protein production of eight insect cell lines.  


A recombinant Autographa californica baculovirus expressing secreted alkaline phosphatase (SEAP) gene was used to evaluate the expression of a secreted glycoprotein in eight insect cell lines derived from Spodoptera frugiperda, Trichoplusia ni, Mamestra brassicae and Estigmene acrea. Because cell density was found to influence protein production, SEAP production was evaluated at optimal cell densities for each cell line on both a per cell and per milliliter basis. On a per cell basis, the T. ni-derived BTI-TN-5B1-4 cells produced a minimum of 20-fold more SEAP than the S. frugiperda-derived Sf9 or Sf2l cell lines and a minimum of 9-fold more than any of the other cell lines growing in serum-containing medium. On a per milliliter basis, BTI-TN-5B1-4 cells produced a minimum of fivefold more SEAP than any of the other cell lines tested. Using cell lines that were adapted to serum-free medium, SEAP yields were the same or better than their counterparts in serum-containing medium. At 3 days postinoculation, extracellular SEAP activity ranged from 59 to 85% of total SEAP activity with cell lines grown in serum-free and serum-containing media. PMID:8314732

Davis, T R; Wickham, T J; McKenna, K A; Granados, R R; Shuler, M L; Wood, H A



Shortest path based splitting line finding for touching cells  

NASA Astrophysics Data System (ADS)

A shortest path based algorithm is proposed in this paper to find splitting lines for touching cells. Firstly, an initial splitting line is obtained through the distance transform of a marker image and the watershed algorithm. Then, the initial splitting line is separated into different line segments if necessary, and the start and end points of these line segments act as the start and end points of shortest path. Finally, the shortest path algorithm is used to find the splitting line between the start and end points, and the final result of touching cells splitting can be formed by the contour of the touching cells and the splitting lines. Experimental results show that the proposed algorithm is efficient for different types of touching cells.

Bai, Xiangzhi; Sun, Changming; Wang, Peng; Zhou, Fugen



Polyamine synthesis in maize cell lines  

SciTech Connect

Uptake of ({sup 14}C)putrescine, ({sup 14}C)arginine, and ({sup 14}C)ornithine was measured in five separate callus cell lines of Zea mays. Each precursor was rapidly taken into the intracellular pool in each culture where, on the average 25 to 50% of the total putrescine was found in a conjugated form, detected after acid hydrolysis. Half-maximal labeling of each culture was achieved in less than 1 minute. Within this time frame of precursor incorporation, only putrescine derived from arginine was conjugated, indicating that putrescine pools derived from arginine may initially be sequestered from ornithine-derived putrescine. The decarboxylase activities were measured in each culture after addition of exogenous polyamine to the growth medium to assess differential regulation of the decarboxylases. Arginine and ornithine decarboxylase activities were augmented by added polyamine, the effect on arginine decarboxylase being eightfold greater than on ornithine decarboxylase. Levels of extractable ornithine decarboxylase were consistently 15- to 100-fold higher than arginine decarboxylase, depending on the titer of extracellular polyamine. Taken as whole the results support the idea that there are distinct populations of polyamine that are initially sequestered after the decarboxylase reactions and that give rise to separate end products and possibly have separate functions.

Hiatt, A. (Scripps Clinic and Research Foundation, La Jolla, CA (USA))



Establishment and characterization of seven human breast cancer cell lines including two triple-negative cell lines.  


Permanently growing cell lines can be invaluable because of their usefulness in a variety of experimental situations. We report the characteristics of seven cell lines designated, SNU-306, SNU-334, SNU-1528, SNU-1553, SNU-1581, SNU-1958 and SNU-2372, which were established from three primary carcinomas, two pleural effusion, one pericardial effusion and one ascitic fluid samples obtained from seven Korean breast carcinoma patients. The histopathology of the primary tumors and their in vitro growth characteristics are described. DNA fingerprinting analysis and genetic alterations in the p53 and EGFR genes were conducted. The expression levels of the ER-?, PR, C-erbB2, E-cadherin, COX-2, MDR and MXR genes were investigated and sensitivity to anticancer drugs was screened. Growth was as adherent cells (four cell lines), floating aggregates (one cell line) and both (two cell lines). All lines were free of mycoplasma or bacteria and were proven unique by DNA fingerprinting analysis using 18 microsatellite markers. Estrogen receptor (ER) mRNA was highly expressed in five cell lines and low or undetectable in SNU-1958 and SNU-2372. Progesterone receptor (PR) mRNA was expressed only in the SNU-306. SNU-1958 and SNU-2372 were hormone receptor-negative and C-erbB2-negative (triple-negative). SNU-1528 had an in-frame deletion of 42 base pairs of p53 gene and showed over 20-fold resistance for taxol compared to the other cell lines. There were no mutation in the EGFR gene; COX-2 was expressed in four cell lines and MXR was expressed in two cell lines. These well-characterized seven breast cancer cell lines, which include two triple-negative cell lines, will be useful for the study of breast cancer biology. PMID:24141649

Ku, Ja-Lok; Park, Sung-Chan; Kim, Kyung-Hee; Jeon, You-Kyung; Kim, Sung-Hee; Shin, Young-Kyoung; Noh, Dong-Young; Im, Seock-Ah; Bang, Yung-Jue; Han, Wonshik; Kim, Woo Ho; Park, Jae-Gahb



Establishment of Mouse Embryonic Stem Cell-Derived Erythroid Progenitor Cell Lines Able to Produce Functional Red Blood Cells  

Microsoft Academic Search

BackgroundThe supply of transfusable red blood cells (RBCs) is not sufficient in many countries. If erythroid cell lines able to produce transfusable RBCs in vitro were established, they would be valuable resources. However, such cell lines have not been established. To evaluate the feasibility of establishing useful erythroid cell lines, we attempted to establish such cell lines from mouse embryonic

Takashi Hiroyama; Kenichi Miharada; Kazuhiro Sudo; Inaho Danjo; Naoko Aoki; Yukio Nakamura; Simon Williams



Authentication of the R06E Fruit Bat Cell Line  

PubMed Central

Fruit bats and insectivorous bats are believed to provide a natural reservoir for a wide variety of infectious diseases. Several lines of evidence, including the successful isolation of infectious viruses, indicate that Marburg virus and Ravn virus have found a major reservoir in colonies of the Egyptian rousette (Rousettus aegyptiacus). To facilitate molecular studies on virus-reservoir host interactions and isolation of viruses from environmental samples, we established cell lines from primary cells of this animal. The cell lines were given to several laboratories until we realized that a contamination with Vero cells in one of the cultures had occurred. Here we describe a general diagnostic procedure for identification of cross-species contamination with the focus on Vero and Rousettus cell lines, and summarize newly discovered properties of the cell lines that may pertain to pathogen discovery.

Jordan, Ingo; Munster, Vincent J.; Sandig, Volker



Topic: Consideration of the Appropriateness of Cell Lines ...  

Center for Biologics Evaluation and Research (CBER)

Text Version... 9:20 History and Characterization of the A3.01 Cell Line Seung Ho Choo and its Tumorigenic Evaluation ... or Cells Derived from Human Tumors ... More results from


Human rhabdomyosarcoma cell lines for rhabdomyosarcoma research: utility and pitfalls.  


Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence. Despite intergroup clinical trials conducted in Europe and North America, outcomes for high risk patients with this disease have not significantly improved in the last several decades, and survival of metastatic or relapsed disease remains extremely poor. Accrual into new clinical trials is slow and difficult, so in vitro cell-line research and in vivo xenograft models present an attractive alternative for preclinical research for this cancer type. Currently, 30 commonly used human RMS cell lines exist, with differing origins, karyotypes, histologies, and methods of validation. Selecting an appropriate cell line for RMS research has important implications for outcomes. There are also potential pitfalls in using certain cell lines including contamination with murine stromal cells, cross-contamination between cell lines, discordance between the cell line and its associated original tumor, imposter cell lines, and nomenclature errors that result in the circulation of two or more presumed unique cell lines that are actually from the same origin. These pitfalls can be avoided by testing for species-specific isoenzymes, microarray analysis, assays for subtype-specific fusion products, and short tandem repeat analysis. PMID:23882450

Hinson, Ashley R P; Jones, Rosanne; Crose, Lisa E S; Belyea, Brian C; Barr, Frederic G; Linardic, Corinne M



Differentiation of Embryonic Stem Cell Lines Generated from Adult Somatic Cells by Nuclear Transfer  

Microsoft Academic Search

Embryonic stem (ES) cells are fully pluripotent in that they can differentiate into all cell types, including gametes. We have derived 35 ES cell lines via nuclear transfer (ntES cell lines) from adult mouse somatic cells of inbred, hybrid, and mutant strains. ntES cells contributed to an extensive variety of cell types, including dopaminergic and serotonergic neurons in vitro and

Teruhiko Wakayama; Viviane Tabar; Ivan Rodriguez; Anthony C. F. Perry; Lorenz Studer; Peter Mombaerts



Survey of Interferon Production and Sensitivity in Human Cell Lines  

PubMed Central

Seven presumed diploid and 11 established cell lines were studied for their ability to produce free interferon in response to a standardized Newcastle disease virus challenge. Interferon production was evaluated in both serum-containing and serum-free medium. The ability of these cell lines to respond to the application of a standard interferon preparation by becoming resistant to virus was also examined. The diploid lines were distinctly more efficient producers of interferon than were the established lines. They also evidenced a greater requirement for serum to produce their maximum titers, but some were able to produce good titers in serum-free medium. The diploid lines were uniformly more sensitive to the application of exogenous interferon than were the established cell lines and attained greater degrees of virus resistance, but all lines tested displayed measurable sensitivity to interferon.

Moehring, J. M.; Stinebring, W. R.; Merchant, D. J.



Caffeine markedly sensitizes human mesothelioma cell lines to pemetrexed  

Microsoft Academic Search

Pemetrexed is a new generation antifolate approved for the treatment of mesothelioma and non-small cell lung cancer. Caffeine\\u000a is known to augment radiation or chemotherapeutic drug-induced cell killing. The current study addresses the impact of caffeine\\u000a on the activity of pemetrexed in mesothelioma cell lines. Caffeine enhanced pemetrexed activity in all four mesothelioma cell\\u000a lines tested (H2052, H2373, H28 and

Sang Hee Min; I. David Goldman; Rongbao Zhao



The transcriptional diversity of 25 Drosophila cell lines  

SciTech Connect

Drosophila melanogaster cell lines are important resources for cell biologists. Here, we catalog the expression of exons, genes, and unannotated transcriptional signals for 25 lines. Unannotated transcription is substantial (typically 19% of euchromatic signal). Conservatively, we identify 1405 novel transcribed regions; 684 of these appear to be new exons of neighboring, often distant, genes. Sixty-four percent of genes are expressed detectably in at least one line, but only 21% are detected in all lines. Each cell line expresses, on average, 5885 genes, including a common set of 3109. Expression levels vary over several orders of magnitude. Major signaling pathways are well represented: most differentiation pathways are ‘‘off’’ and survival/growth pathways ‘‘on.’’ Roughly 50% of the genes expressed by each line are not part of the common set, and these show considerable individuality. Thirty-one percent are expressed at a higher level in at least one cell line than in any single developmental stage, suggesting that each line is enriched for genes characteristic of small sets of cells. Most remarkable is that imaginal discderived lines can generally be assigned, on the basis of expression, to small territories within developing discs. These mappings reveal unexpected stability of even fine-grained spatial determination. No two cell lines show identical transcription factor expression. We conclude that each line has retained features of an individual founder cell superimposed on a common ‘‘cell line‘‘ gene expression pattern. Wereport the transcriptional profiles of 25 Drosophila melanogaster cell lines, principally by whole-genome tiling microarray analysis of total RNA, carried out as part of the modENCODE project. The data produced in this study add to our knowledge of the cell lines and of the Drosophila transcriptome in several ways. We summarize the expression of previously annotated genes in each of the 25 lines with emphasis on what those patterns reveal about the origins of the lines and the stability of spatial expression patterns. We also offer an initial analysis of previously unannotated transcripts in the cell lines.

Cherbas, Lucy; Willingham, Aarron; Zhang, Dayu; Yang, Li; Zou, Yi; Eads, Brian D.; Carlson, Joseph W.; Landolin, Jane M.; Kapranov, Philipp; Dumais, Jacqueline; Samsonova, Anastasia; Choi, Jeong-Hyeon; Roberts, Johnny; Davis, Carrie A.; Tang, Haixu; van Baren, Marijke J.; Ghosh, Srinka; Dobin, Alexander; Bell, Kim; Lin, Wei; Langton, Laura; Duff, Michael O.; Tenney, Aaron E.; Zaleski, Chris; Brent, Michael R.; Hoskins, Roger A.; Kaufman, Thomas C.; Andrews, Justen; Graveley, Brenton R.; Perrimon, Norbert; Celniker, Susan E.; Gingeras, Thomas R.; Cherbas, Peter



Using Neuroblastoma Cell Lines to Examine Organophosphate Neurotoxicity.  

National Technical Information Service (NTIS)

The paper describes the initial characterization of neuroblastoma cell lines to address several aspects of organophosphate neurotoxicity. Several commercially available human and mouse cell lines (i.e., SY5Y, IMR-32, SK-N-MC, NB41A3) were evaluated for th...

B. Veronesi M. Ehrich



A comparison of the cell lines used in meningioma research  

Microsoft Academic Search

BackgroundImmortal cell lines and cell lines derived from operative specimens transplanted into animal models are used in meningioma research. We address 2 criticisms of the mouse xenograft flank tumor model: Why are tumor induction rates derived from operative specimens low and inconsistent? Are flank tumors meningiomas?

Brian T. Ragel; William T. Couldwell; David L. Gillespie; Merideth M. Wendland; Kum Whang; Randy L. Jensen



Development and characterization of rabbit proximal tubular epithelial cell lines  

Microsoft Academic Search

Development and characterization of rabbit proximal tubular epithelial cell lines. We have isolated rabbit kidney proximal tubular epithelial cell lines. The selection was based on their ability to form confluent monolayers on porous supports and to maintain receptor-mediated signal transduction and ion transport, characteristic of the proximal tubule. The isolation method consisted of several steps: (1) superficial cortical proximal tubule

Michael F Romero; Janice G Douglas; Richard L Eckert; Ulrich Hopfer; James W Jacobberger



Permissiveness of human hepatoma cell lines for HCV infection  

PubMed Central

Background Although primary and established human hepatoma cell lines have been evaluated for hepatitis C virus (HCV) infection in vitro, thus far only Huh7 cells have been found to be highly permissive for infectious HCV. Since our understanding of the HCV lifecycle would benefit from the identification of additional permissive cell lines, we assembled a panel of hepatic and non-hepatic cell lines and assessed their ability to support HCV infection. Here we show infection of the human hepatoma cell lines PLC/PRF/5 and Hep3B with cell culture-derived HCV (HCVcc), albeit to lower levels than that achieved in Huh7 cells. To better understand the reduced permissiveness of PLC and Hep3B cells for HCVcc infection, we performed studies to evaluate the ability of each cell line to support specific steps of the viral lifecycle (i.e. entry, replication, egress and spread). Results We found that while the early events in HCV infection (i.e. entry plus replication initiation) are cumulatively equivalent or only marginally reduced in PLC and Hep3B cells, later steps of the viral life cycle such as steady-state replication, de novo virus production and/or spread are impaired to different degrees in PLC and Hep3B cultures compared to Huh7 cell cultures. Interestingly, we also observed that interferon stimulated gene (i.e. ISG56) expression was significantly and differentially up-regulated in PLC and Hep3B cells following viral infection. Conclusions We conclude that the restrictions observed later during HCV infection in these cell lines could in part be attributed to HCV-induced innate signaling. Nevertheless, the identification of two new cell lines capable of supporting authentic HCVcc infection, even at reduced levels, expands the current repertoire of cell lines amendable for the study of HCV in vitro and should aid in further elucidating HCV biology and the cellular determinants that modulate HCV infection.



Production of Uniparental Embryonic Stem Cell Lines  

Microsoft Academic Search

\\u000a Embryonic stem cells, or induced pluripotent cells derived from somatic cells, can yield differentiated progeny with potential\\u000a applicability for tissue repair. This chapter describes the generation of embryonic stem cells from gamete-derived uniparental\\u000a embryos. These embryonic stem cells can be patient-derived and potentially histocompatible with the gamete donor. The production\\u000a of uniparental embryos followed by derivation of embryonic stem cells

Sigrid Eckardt; K. John McLaughlin


Cell-cell signaling interactions coordinate multiple cell behaviors that drive morphogenesis of the lateral line  

PubMed Central

The zebrafish sensory lateral line system has emerged as a powerful model for the mechanistic study of collective cell migration and morphogenesis. Recent work has uncovered the details of a signaling network involving the Wnt/?-catenin, Fgf and Delta-Notch pathways that patterns the migrating lateral line primordium into distinct regions. Cells within these regions exhibit different fundamental behaviors that together orchestrate normal lateral line morphogenesis. In this review, we summarize the signaling network that patterns the migrating lateral line primordium and describe how this patterning coordinates crucial morphogenic cell behaviors.

Aman, Andy



Establishment and culture of leukemia-lymphoma cell lines.  


The advent of continuous human leukemia-lymphoma cell lines as a rich resource of abundant, accessible, and manipulable living cells has contributed significantly to a better understanding of the pathophysiology of hematopoietic tumors. The first leukemia-lymphoma cell lines were established in 1963 and since then large numbers of new cell lines have been described. The major advantages of continuous leukemia-lymphoma cell lines are the unlimited supply and worldwide availability of identical cell material and the infinite viable storability in liquid nitrogen. These cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro under preservation of most cellular features, and by specific genetic alterations. Here some of the more promising techniques for establishing new leukemia-lymphoma cell lines and the basic principles for culturing these cells are described. Several clinical and cell culture parameters might have some influence on the success rate, e.g., choice of culture medium and culture conditions, specimen site of the primary cells, and status of the patient at the time of sample collection. PMID:21516408

Drexler, Hans G



GREG cells, a dysferlin-deficient myogenic mouse cell line  

PubMed Central

The dysferlinopathies (e.g. LGMD2b, Myoshi myopathy) are progressive, adult-onset muscle wasting syndromes caused by mutations in the gene coding for dysferlin. Dysferlin is a large (~200 kDa) membrane-anchored protein, required for maintenance of plasmalemmal integrity in muscle fibers. To facilitate analysis of dysferlin function in muscle cells, we have established a dysferlin-deficient myogenic cell line (GREG cells) from the A/J mouse, a genetic model for dysferlinopathy. GREG cells have no detectable dysferlin expression, but proliferate normally in growth medium and fuse into functional myotubes in differentiation medium. GREG myotubes exhibit deficiencies in plasma membrane repair, as measured by laser wounding in the presence of FM1-43 dye. Under the wounding conditions used, the majority (~66%) of GREG myotubes lack membrane repair capacity, while no membrane repair deficiency was observed in dysferlin-normal C2C12 myotubes, assayed under the same conditions. We discuss the possibility that the observed heterogeneity in membrane resealing represents genetic compensation for dysferlin deficiency.

Humphrey, Glen W.; Mekhedov, Elena; Blank, Paul S.; de Morree, Antoine; Pekkurnaz, Gulcin; Nagaraju, Kanneboyina; Zimmerberg, Joshua



Isolation of Amniotic Stem Cell Lines With Potential for Therapy  

Microsoft Academic Search

Stem cells capable of differentiating to multiple lineages may be valuable for therapy. We report the isolation of human and rodent amniotic fluid-derived stem (AFS) cells that express embryonic and adult stem cell markers. Undifferentiated AFS cells expand extensively without feeders, double in 36 h and are not tumorigenic. Lines maintained for over 250 population doublings retained long telomeres and

Paolo De Coppi; Georg Bartsch; M Minhaj Siddiqui; Tao Xu; Cesar C. Santos; Laura Perin; Gustavo Mostoslavsky; Evan Y. Snyder; James J. Yoo; Mark E. Furth; Shay Soker; Anthony Atala



Human Embryonic Stem Cell Lines Derived from Discarded Embryos  

Microsoft Academic Search

ABSTRACT Human pluripotent embryonic stem (ES) cells have important potential in regenerative medicine and as models for human preimplantation development; how- ever, debate continues over whether embryos should be destroyed to produce human ES cells. We have derived four ES cell lines on mouse embryonic fibroblast cells in medium supplemented with basic fibroblast growth fac- tor, human recombinant leukemia inhibitory

Maisam Mitalipova; John Calhoun; Soojung Shin; David Wininger; Thomas Schulz; Scott Noggle; Alison Venable; Ian Lyons; Allan Robins; Steven Stice



Respiratory epithelial cell lines exposed to anoxia produced inflammatory mediator  

PubMed Central

Human epithelial cell lines were utilized to examine the effects of anoxia on cellular growth and metabolism. Three normal human epithelial cells lines (A549, NHBE, and BEAS-2B) as well as a cystic fibrosis cell line (IB3-1) and its mutation corrected cell line (C38) were grown in the presence and absence of oxygen for varying periods of time. Interleukin-8 (IL-8) levels were measured by enzyme-linked immunosorbent assay technique. Cellular metabolism and proliferation were assayed by determining mitochondrial oxidative burst activity by tetrazolium compound reduction. The viability of cells was indirectly measured by lactate dehydrogenase release. A549, NHBE, and BEAS-2B cells cultured in the absence of oxygen showed a progressive decrease in metabolic activity and cell proliferation after one to three days. There was a concomitant increase in IL-8 production. Cell lines from cystic fibrosis (CF) patients did not show a similar detrimental effect of anoxia. However, the IL-8 level was significantly increased only in IB3-1 cells exposed to anoxia after two days. Anoxia appears to affect certain airway epithelial cell lines uniquely with decreased cellular proliferation and a concomitant increased production of a cytokine with neutrophilic chemotactic activity. The increased ability of the CF cell line to respond to anoxia with increased secretion of inflammatory cytokines may contribute to the inflammatory damage seen in CF bronchial airway. This study indicates the need to use different cell lines in in vitro studies investigating the role of epithelial cells in airway inflammation and the effects of environmental influences.

Shahriary, Cyrus M.; Nussbaum, Eliezer



Plant cell biology through the window of the highly synchronized tobacco BY2 cell line  

Microsoft Academic Search

Synchronous cell systems are highly desirable for investigating various aspects of plant cell biology. However, to date, the tobacco BY-2 cell line is the only plant cell line which can be synchronized to high levels. A cell synchrony starting from S phase is obtained after release of BY-2 cells from aphidicolin treatment, while that from M phase is available after

Toshiyuki Nagata; Fumi Kumagai



Genotyping of 73 UM-SCC head and neck squamous cell carcinoma cell lines  

PubMed Central

Background We established multiple UM-SCC (University of Michigan Squamous Cell Carcinoma) cell lines. With time, these have been distributed to other labs all over the world. Recent scientific discussions have noted the need to confirm the origin and identity of cell lines in grant proposals and journal articles. We genotyped the UM-SCC cell lines in our collection to confirm their unique identity. Design Early passage UM-SCC cell lines were genotyped and photographed. Results Thus far, 73 unique head and neck UM-SCC cell lines (from 65 donors including 21 lines from 17 females) were genotyped. In 7 cases separate cell lines were established from the same donor. Conclusions These results will be posted on the U of M Head and Neck SPORE Tissue Core website for other investigators to confirm that the UM-SCC cells used in their laboratories have the correct features. Publications using UM-SCC cell lines should confirm the genotype.

Brenner, J. Chad; Graham, Martin P.; Kumar, Bhavna; Saunders, Lindsay M.; Kupfer, Robbi; Lyons, Robert H.; Bradford, Carol R.; Carey, Thomas E.



Establishment of Human Colon Cancer Cell Lines from Fresh Tumors versus Xenografts: Comparison of Success Rate and Cell Line Features  

Microsoft Academic Search

Obtaining representative human colon cancer cell lines from fresh tumors is technically difficult. Using 32 tumor fragments from patients with colon cancer, the present study shows that prior xenograft leads to more efficient cell line establishment compared with direct establishment from fresh tumors (P < 0.05). From 26 tumor specimens, we successfully established 20 tumor xenografts in nude mice (77%);

Virginie Dangles-Marie; Marc Pocard; Sophie Richon; Louis-Bastien Weiswald; Jean-Gabriel Judde; Jean-Louis Janneau; Nathalie Auger; Pierre Validire; Bernard Dutrillaux; Francoise Praz; Dominique Bellet; Marie-France Poupon; Departement de Biologie



Expression pattern of galectin-3 in neural tumor cell lines.  


Galectin-3 is a member of the galectin family of beta-galactoside-specific animal lectins. Here we show that galectin-3 is constitutively expressed in 15 out of 16 glioma cell lines tested, but not by normal or reactive astrocytes, oligodendrocytes, glial O-2A progenitor cells and the oligodendrocyte precursor cell line Oli-neu. Galectin-3 is also expressed by one oligodendroglioma cell line, but not by primitive neuroectodermal tumor and 4 neuroblastoma cell lines tested so far. In all galectin-3 expressing cell lines, the lectin is predominantly, if not exclusively, localized intracellularly and carries an active carbohydrate recognition domain (shown for C6 rat glioma cells). Moreover, in contrast to primary astrocytes, glioma cells do not or only weakly adhere to substratum-bound galectin-3, probably reflecting an unusual glycosylation pattern. Our findings indicate that the expression of galectin-3 selectively correlates with glial cell transformation in the central nervous system and could thus serve as a marker for glial tumor cell lines and glial tumors. PMID:10723067

Kuklinski, S; Pesheva, P; Heimann, C; Urschel, S; Gloor, S; Graeber, S; Herzog, V; Pietsch, T; Wiestler, O D; Probstmeier, R



Differential effects of bisphosphonates on breast cancer cell lines.  


Bisphosphonates may induce direct anti-tumor effects in breast cancer cells in vitro. In this study, six bisphosphonates were administered to three breast cancer cell lines. Cell proliferation was measured by quantification of the expression of Cyclin D1 mRNA. Apoptosis was determined by flow cytometry of a DNA fragmentation assay. We demonstrated that bisphosphonates have direct effects on cell proliferation and apoptosis in different breast cancer cell lines. However, not all bisphosphonates act equally on breast cancer cells in vitro. Zoledronate seems to be the most potent of the six bisphosphonates. This in vitro study showed that bisphosphonates possess promising anti-tumor potential. PMID:16621245

Verdijk, R; Franke, H R; Wolbers, F; Vermes, I



Novel human bronchial epithelial cell lines for cystic fibrosis research  

PubMed Central

Immortalization of human bronchial epithelial (hBE) cells often entails loss of differentiation. Bmi-1 is a protooncogene that maintains stem cells, and its expression creates cell lines that recapitulate normal cell structure and function. We introduced Bmi-1 and the catalytic subunit of telomerase (hTERT) into three non-cystic fibrosis (CF) and three ?F508 homozygous CF primary bronchial cell preparations. This treatment extended cell life span, although not as profoundly as viral oncogenes, and at passages 14 and 15, the new cell lines had a diploid karyotype. Ussing chamber analysis revealed variable transepithelial resistances, ranging from 200 to 1,200 ?·cm2. In the non-CF cell lines, short-circuit currents were stimulated by forskolin and inhibited by CFTR(inh)-172 at levels mostly comparable to early passage primary cells. CF cell lines exhibited no forskolin-stimulated current and minimal CFTR(inh)-172 response. Amiloride-inhibitable and UTP-stimulated currents were present, but at lower and higher amplitudes than in primary cells, respectively. The cells exhibited a pseudostratified morphology, with prominent apical membrane polarization, few apoptotic bodies, numerous mucous secretory cells, and occasional ciliated cells. CF and non-CF cell lines produced similar levels of IL-8 at baseline and equally increased IL-8 secretion in response to IL-1?, TNF-?, and the Toll-like receptor 2 agonist Pam3Cys. Although they have lower growth potential and more fastidious growth requirements than viral oncogene transformed cells, Bmi-1/hTERT airway epithelial cell lines will be useful for several avenues of investigation and will help fill gaps currently hindering CF research and therapeutic development.

Fulcher, M. L.; Gabriel, S. E.; Olsen, J. C.; Tatreau, J. R.; Gentzsch, M.; Livanos, E.; Saavedra, M. T.; Salmon, P.; Randell, S. H.



Recent developments on human cell lines for the bioartificial liver.  


Most bioartificial liver (BAL) devices contain porcine primary hepatocytes as their biological component. However, alternatives are needed due to xenotransplantation associated risks. Human liver cell lines have excellent growth characteristics and are therefore candidates for application in BAL devices. Tumour-derived cell lines HepG2 and C3A express a variety of liver functions, but some specific liver functions, like ammonia detoxification and ureagenesis are insufficient. Immortalised human hepatocytes might offer better prospects. The balance between immortalisation and transformation with dedifferentiation of cells seems controllable by conditional immortalisation and/or the use of telomerase as immortalising agent. Another promising approach will be the use of embryonic or adult human stem cells. Rodent stem cells have been directed to hepatic differentiation in vitro, which might be applicable to human stem cells. However, both functionality and safety of immortalised human liver cell lines and differentiated stem cells should be improved before successful use in BAL devices becomes reality. PMID:11999190

Hoekstra, R; Chamuleau, R A F M



Steroid hormone receptors in three human gastric cancer cell lines  

Microsoft Academic Search

Steroid hormone receptors in three human gastric adenocarcinoma cell lines and their transplanted tumors (except nontumorigenic KATO-III) in nude mice were determined by dextran-coated charcoal assay. Progesterone receptors (PgR) were found in all cell lines, transplanted NUGC-3, and AZ 521 tumors. Estrogen receptors (ER) were found in KATO-III cells, transplanted NUGC-3, and AZ 521 tumors, whereas glucocorticoid receptors (GR) were

Chew-Wun Wu; Yuh-Fang Chang; Tsuey-Hwa Yeh; Tai-Jay Chang; Wing-Yiu Lui; Fang-Ku P'eng; Chin-Wen Chi



Epithelial mesenchymal transition traits in human breast cancer cell lines  

Microsoft Academic Search

Epithelial mesenchymal transition (EMT) has long been associated with breast cancer cell invasiveness and evidence of EMT\\u000a processes in clinical samples is growing rapidly. Genome-wide transcriptional profiling of increasingly larger numbers of\\u000a human breast cancer (HBC) cell lines have confirmed the existence of a subgroup of cell lines (termed Basal B\\/Mesenchymal)\\u000a with enhanced invasive properties and a predominantly mesenchymal gene

T. Blick; E. Widodo; H. Hugo; M. Waltham; M. E. Lenburg; R. M. Neve; E. W. Thompson



SK HEP1: A human cell line of endothelial origin  

Microsoft Academic Search

Summary  SK-HEP-1 is an immortal, human cell line derived from the ascitic fluid of a patient with adenocarcinoma of the liver. We\\u000a have determined that these cells are of endothelial origin. Despite the location of the tumor from which SK HEP-1 was derived,\\u000a the cell line does not have properties of hepatocytes. Northern blot analysis of total cellular RNA shows no

Sue C. Heffelfinger; Hal H. Hawkins; Jim Barrish; Linda Taylor; Gretchen J. Darlington



Hepatitis C virus infection of neuroepithelioma cell lines  

PubMed Central

Background & Aims Hepatitis C virus (HCV) establishes chronic infections in 3% of the world's population. Infection leads to progressive liver disease; hepatocytes are the major site of viral replication in vivo. However, chronic infection is associated with a variety of extrahepatic syndromes, including central nervous system (CNS) abnormalities. We therefore screened a series of neural and brain-derived cell lines for their ability to support HCV entry and replication. Methods We used a panel of neural-derived cell lines, HCV pseudoparticles (HCVpp), and an infectious, HCV JFH-1 cell-culture system (HCVcc) to assess viral tropism. Results Two independently derived neuroepithelioma cell lines (SK-N-MC and SK-PN-DW) permitted HCVpp entry. In contrast, several neuroblastoma, glioma, and astrocytoma cell lines were refractory to HCVpp infection. HCVcc infected the neuroepithelioma cell lines and established a productive infection. Permissive neuroepithelioma cells expressed CD81, scavenger receptor BI (SR-BI), and the tight junction proteins Claudin-1 (CLDN1) and occludin, whereas non-permissive neural cell lines lacked CLDN1 and in some cases SR-BI. HCVpp infection of the neuroepithelioma cells was neutralized by antibodies to CD81, SR-BI, CLDN1 and HCV E2. Furthermore, anti-CD81, interferon and the anti-NS3 protease inhibitor VX-950 significantly reduced HCVcc infection of neuroepithelioma and hepatoma cells. Conclusions Neuroepithelioma-derived cell lines express functional receptors that support HCV entry at comparable levels to that of hepatoma cells. HCV infection in vitro is not restricted to hepatic-derived cells, so HCV might infect cells of the CNS in vivo.

Fletcher, Nicola F; Yang, Jian Ping; Farquhar, Michelle J; Hu, Ke; Davis, Christopher; He, Qiuchen; Dowd, Kimberly; Ray, Stuart C; Krieger, Sophie E; Neyts, Johan; Baumert, Thomas F; Balfe, Peter; McKeating, Jane A; Wong-Staal, Flossie



Cell line models for differentiation: preadipocytes and adipocytes.  


In vitro models have been invaluable in determining the mechanisms involved in adipocyte proliferation, differentiation, adipokine secretion and gene/protein expression. The cells presently available for research purposes all have unique advantages and disadvantages that one should be aware of when selecting cells. Established cell lines, such as 3T3-L1 cells, are easier and less costly to use than freshly isolated cells, even though freshly isolated cells allow for various comparisons such as the in vitro evaluation of different in vivo conditions that may not be possible using cell lines. Moreover, stem cells, transdifferentiated cells or dedifferentiated cells are relatively new cell models being evaluated for the study of adipocyte regulation and physiology. The focus of this brief review is to highlight similarities and differences in adipocyte models to aid in appropriate model selection and data interpretation for successful advancement of our understanding of adipocyte biology. PMID:20864461

Poulos, Sylvia P; Dodson, Michael V; Hausman, Gary J



Hybridization between T and B lymphoma cell lines.  

PubMed Central

The AKR thymoma line BW 5147 has been successfully hybridized with the IgM-bearing (BALB/c x NZB)F1 B lymphoma line WEHI 231. In the hybrids formed, T-cell characteristics were dominant, i.e. there was no expression of IgM but continued expression of Thy-1.1 in eight out of eight lines. Moreover, in two out of eight lines, the Thy-1.2 allele was also expressed. We conclude that, in its ability to hybridize, BW 5147 is not restricted to cells of similar ontogenetic origin (i.e. T cells) and that fusion with the thymoma can lead to suppression of B-cell gene expression and derepression of genes for T-cell markers.

Taussig, M J; Holliman, A; Wright, L J



Development of a cell line from Echinococcus granulosus germinal layer.  


In vitro culture of parasitic helminths provides an important tool to study cell regeneration and physiology, as well as for molecular biology and genetic engineering studies. In the present study, we established in vitro propagation of cells from Echinococcus granulosus germinal cyst layer. E. granulosus germinal cells grew beyond 100 passages and showed no signs of reduced proliferation capacity. Microscopic analysis revealed that cells grew both attached to the substrate and in suspension, forming three-dimensional structures like mammalian stem cell aggregates. Examination of the chromosome number of attached germinal cells showed a high degree of heteroploidy, suggesting the occurrence of transformation during culture. Monolayer cells survived cryopreservation and were able to proliferate after thawing. Based on the characteristics displayed by E. granulosus germinal cells, we establish a cell line from the E. granulosus germinal layer. Furthermore, we propose that this cell line could be useful for drug screening and for obtaining parasite material. PMID:23860182

Albani, Clara María; Cumino, Andrea Carina; Elissondo, María Celina; Denegri, Guillermo María



[Decontamination of continual cell lines spontaneously infected with mycoplasmas].  


The continual cell lines of bovine kidneys MDBK and AUBEK, and porcine kidneys RPD and IBRS, spontaneously infected with Mycoplasma arginini and Acholeplasma laidlawii, were decontaminated by the method of selective elimination. Two elimination procedures were modified to be used for the decontamination: one based on the reduction of infection by the light treatment of the cultures, the other based on the selection of mycoplasma-free cell population through cell clonation. On the basis of a long-continued control of the cell clones a methodical procedure of the preparation of mycoplasma-free cell lines was worked out. PMID:3090766

Machatková, M; Jurmanová, K; Snejdar, V



Human papillomavirus in vulvar and vaginal carcinoma cell lines.  

PubMed Central

A number of reports associate human papillomavirus (HPV) with cervical cancer and cancer cell lines derived from this tumour type. Considerably fewer reports have focused on the role of HPV in carcinomas from other sites of female anogenital squamous epithelia. In this study we have tested for the presence of HPV in eight low-passage vulvar carcinoma cell lines and one extensively passaged cell line, A431. One cell line from a primary vaginal carcinoma was included. The presence of the HPV was evaluated by the polymerase chain reaction (PCR), by Southern blot analysis and by two-dimensional gel electrophoresis. General primer-mediated PCR was applied by using primers from the L1 region, E1 region and HPV 16 E7 region. Southern blot hybridisation was performed under low-stringency conditions (Tm = -35 degrees C) using a whole genomic HPV 6/16/18 probe mixture and under high stringency conditions (Tm = -18 degrees C) with the whole genomic probes of HPV 16 and 33. HPV 16 E6-E7 mRNA was assessed by ribonuclease protection assay (RPA). HPV was found in only one vulvar carcinoma cell line, UM-SCV-6. The identified type, HPV 16, was integrated in the cell genome and could be amplified with all primers used. Also E6-E7 transcripts were found in these cells. Five original tumour biopsies were available from the HPV-negative cell lines for in situ hybridisation. All these were HPV negative with both the HPV 6/16/18 screening probe mixture under low stringency and the HPV 16 probe under high stringency. The results indicate that vulvar carcinoma cell lines contain HPV less frequently than cervical carcinoma cell lines and suggest that a significant proportion of vulvar carcinomas may evolve by an HPV-independent mechanism. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6

Hietanen, S.; Grenman, S.; Syrjanen, K.; Lappalainen, K.; Kauppinen, J.; Carey, T.; Syrjanen, S.



Establishment of a corneal epithelial cell line spontaneously derived from human limbal cells  

Microsoft Academic Search

The objective of this study was to establish a spontaneously derived human corneal epithelial cell line from a normal human limbus that retains differentiation potential and proliferative properties under continuous cell culture. After 50 passages of epithelial cells obtained from human limbal tissue a cell line spontaneously emerged. The immortalized cells showed a cobblestone appearance and displayed dense microvilli on

Jingbo liu; Ge Song; Zhichong Wang; Bing Huang; Qianying Gao; Bingqian Liu; Ying Xu; Xuanwei Liang; Ping Ma; Nan Gao; Jian Ge



Isolation and Growth of Prostate Stem Cells and Establishing Cancer Cell Lines from Human Prostate Tumors.  

National Technical Information Service (NTIS)

The objective of this proposal was to isolate, grow, and characterize normal prostate stem cells and establish new prostate cancer cell lines from fresh human prostate tissues. The hypothesis is that prostate stem cells express defined stem cell markers, ...

D. V. Griend



Susceptibilities of medaka (Oryzias latipes) cell lines to a betanodavirus  

PubMed Central

Background Betanodaviruses, members of the family Nodaviridae, have bipartite, positive-sense RNA genomes and are the causal agents of viral nervous necrosis in many marine fish species. Recently, the viruses were shown to infect a few freshwater fish species including a model fish medaka (Oryzias latipes). Although virological study using cultured medaka cells would provide a lot of insight into virus-fish interactions in molecular aspects, no such cells have yet been tested for virus susceptibility. Results We tested ten medaka cell lines for susceptibilities to redspotted grouper nervous necrosis virus (RGNNV). Although the viral coat protein was detected in all the cell lines inoculated, the levels of cytopathic effect development and viral propagation were quite different among the cell lines. Those levels were especially high in OLHNI-1 and OLHNI-2 cells, but were extremely low in OLME-104 cells. Some cell lines entered into antiviral state after RGNNV infections probably because of inducing an antiviral system. This is the first report to examine the susceptibilities of cultured medaka cells against a virus. Conclusion OLHNI-1 and OLHNI-2 cells are candidates of new standard cells for betanodavirus study because of their high susceptibilities to the virus and their several advantages as model fish cells.



Establishment and characterization of a chicken mononuclear cell line.  


A new chicken mononuclear cell line (MQ-NCSU) has been established. The starting material used to initiate this cell line was a transformed spleen from a female Dekalb XL chicken which had been experimentally challenged with the JM/102W strain of the Marek's disease virus. After homogenization, a single cell suspension of splenic cells was cultured using L.M. Hahn medium supplemented with 10 microM 2-mercaptoethanol. Under these culture conditions, a rapidly proliferating cell was observed and then expanded after performing limiting dilution cultures. These cells were moderately adherent and phagocytic for sheep red blood cells and Salmonella typhimurium. When tested against a panel of monoclonal antibodies (mAb) using the flow cytometry, MQ-NCSU cells stained readily with anti-chicken monocyte specific (K-1) mAb but did not stain with mAb detecting T-helper, T-cytotoxic/suppressor, and NK cells. MQ-NCSU cells expressed very high levels of Ia antigens and transferrin receptors. In addition, cell-free supernatant obtained from MQ-NCSU culture contained a factor which exhibited cytolytic activity against tumor cell targets. Based on their cultural, morphological, and functional characteristics and mAb reactivity profile, we conclude that MQ-NCSU cell line represents a malignantly-transformed cell which shares features characteristic of cells of the mononuclear phagocyte lineage. PMID:2176014

Qureshi, M A; Miller, L; Lillehoj, H S; Ficken, M D



Phenotypes and Karyotypes of Human Malignant Mesothelioma Cell Lines  

PubMed Central

Background Malignant mesothelioma is an aggressive tumour of serosal surfaces most commonly pleura. Characterised cell lines represent a valuable tool to study the biology of mesothelioma. The aim of this study was to develop and biologically characterise six malignant mesothelioma cell lines to evaluate their potential as models of human malignant mesothelioma. Methods Five lines were initiated from pleural biopsies, and one from pleural effusion of patients with histologically proven malignant mesothelioma. Mesothelial origin was assessed by standard morphology, Transmission Electron Microscopy (TEM) and immunocytochemistry. Growth characteristics were assayed using population doubling times. Spectral karyotyping was performed to assess chromosomal abnormalities. Authentication of donor specific derivation was undertaken by DNA fingerprinting using a panel of SNPs. Results Most of cell lines exhibited spindle cell shape, with some retaining stellate shapes. At passage 2 to 6 all lines stained positively for calretinin and cytokeratin 19, and demonstrated capacity for anchorage-independent growth. At passage 4 to 16, doubling times ranged from 30–72 hours, and on spectral karyotyping all lines exhibited numerical chromosomal abnormalities ranging from 41 to 113. Monosomy of chromosomes 8, 14, 22 or 17 was observed in three lines. One line displayed four different karyotypes at passage 8, but only one karyotype at passage 42, and another displayed polyploidy at passage 40 which was not present at early passages. At passages 5–17, TEM showed characteristic features of mesothelioma ultrastructure in all lines including microvilli and tight intercellular junctions. Conclusion These six cell lines exhibit varying cell morphology, a range of doubling times, and show diverse passage-dependent structural chromosomal changes observed in malignant tumours. However they retain characteristic immunocytochemical protein expression profiles of mesothelioma during maintenance in artificial culture systems. These characteristics support their potential as in vitro model systems for studying cellular, molecular and genetic aspects of mesothelioma.

Relan, Vandana; Morrison, Leanne; Parsonson, Kylie; Clarke, Belinda E.; Duhig, Edwina E.; Windsor, Morgan N.; Matar, Kevin S.; Naidoo, Rishendran; Passmore, Linda; McCaul, Elizabeth; Courtney, Deborah; Yang, Ian A.; Fong, Kwun M.; Bowman, Rayleen V.



Cadherin-11 is expressed in invasive breast cancer cell lines.  


In several cancers, including breast cancer, loss of E-cadherin expression is correlated with a loss of the epithelial phenotype and with a gain of invasiveness. Cells that have lost E-cadherin expression are either poorly invasive with a rounded phenotype, or highly invasive, with a mesenchymal phenotype. Most cells lacking E-cadherin still retain weak calcium-dependent adhesion, indicating the presence of another cadherin family member. We have now examined the expression of the mesenchymal cadherin, cadherin-11, in breast cancer cell lines. Cadherin-11 mRNA and protein, as well as a variant form, are expressed in the most invasive cell lines but not in any of the noninvasive cell lines. Cadherin-11 is localized to a detergent-soluble pool and is associated with both alpha- and beta-catenin. Immunocytochemistry shows that cadherin-11 is localized to the cell membrane at sites of cell-cell contact as well as at lamellipodia-like projections, which do not interact with other cells. These results suggest that cadherin-11 expression may be well correlated with the invasive phenotype in cancer cells and may serve as a molecular marker for the more aggressive, invasive subset of tumors. Cadherin-11 may mediate the interaction between malignant tumor cells and other cell types that normally express cadherin-11, such as stromal cells or osteoblasts or perhaps even with the surrounding extracellular matrix, thus facilitating tumor cell invasion and metastasis. PMID:10029089

Pishvaian, M J; Feltes, C M; Thompson, P; Bussemakers, M J; Schalken, J A; Byers, S W




EPA Science Inventory

THIS ABSTRACT WAS SUBMITTED ELECTRONICALLY;. SPACE CONSTRAINTS WERE SEVERE) Methylation of Arsenite by Some Mammalian Cell Lines. Methylation of arsenite is thought to play an important role in the carcinogenicity of arsenic. Aim 1: Determine if there is diffe...



Microsoft Academic Search

PurposeTo examine the cytokine profile of epithelial cells lining the human urinary tract with the aim of differentiating between the constitutive and disease-related cytokine production in these tissues.




Cell penetrating peptide conjugated bioreducible polymer for siRNA delivery  

Microsoft Academic Search

The primary cardiomyocyte–specific peptide (PCM) and the cell-penetrating peptide (CPP), HIV-Tat (49–57), were incorporated into the polymer, cystamine bisacrylamide-diaminohexane (CBA-DAH), to increase the delivery of RNAi to target cells, specifically cardiomyocytes. Interestingly, the impact of PCM and Tat conjugation on cellular uptake and transfection efficiency was greater in H9C2 rat cardiomyocytes than in NIH 3T3 cells. We examined the potential

Hye Yeong Nam; Jaesung Kim; Soojin Kim; James W. Yockman; Sung Wan Kim; David A. Bull



Human oesophageal adenocarcinoma cell lines JROECL 47 and JROECL 50 are admixtures of the human colon carcinoma cell line HCT 116  

Microsoft Academic Search

In two recently described human oesophageal adenocarcinoma cell lines JROECL 47 and JROECL 50, derived from one tumour, we detected identical E-cadherin and ?-catenin gene mutations as in colon carcinoma cell line HCT 116. We demonstrate by HLA-typing, mutation analysis and microsatellite analysis that cell lines JROECL 47 and JROECL 50 are admixtures of the human colon adenocarcinoma cell line

B P L Wijnhoven; M G J Tilanus; A G Morris; S J Darnton; H W Tilanus; W N M Dinjens



Phosphorylation of Pyruvate Kinase and Glycolytic Metabolism in Three Human Glioma Cell Lines  

Microsoft Academic Search

Three cell lines established from human gliomas were found to differ in the capacity to phosphorylate the glycolytic enzyme pyruvate kinase in vitro. Phosphorylation in the glioblastoma cell line U-138 was more pronounced than in the glioma cell line Hs 683 and in the glioblastoma cell line A-172. All 3 cell lines showed similar pyruvate kinase isozyme patterns and expressed

Paschal A. Oude Weernink; Gert Rijksen; Gerard E. J. Staal




Microsoft Academic Search

Introduction: Zamzam water is unique in its natural characteristics as zamzam water has a strong anti-inflammatory and it caused downregulation of genes after induction of colon tumor in rate, we test this hypothesis on human by studying the cell lines from uterine fibro-chondrosarcoma. Material and methods: Uterine fibrochondrosarcoma cell line were incubated with zamzam water 20 c.c. for one week

Ali Farid Mohammed Ali; Ermilando Cosemi; Sayed Kamel; Sana Mohammed; Maged Elhefnawy; Laila Farid; Samer Shaker


Alkylating Agent Resistance: In vitro Studies with Human Cell Lines  

Microsoft Academic Search

Development of in vitro resistance to HN2 (also called mustargen or mechlorethamine hydrochloride), N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU), and cisplatin [cis-diamminedichloroplatinum(II)] was achieved in two human cell lines, the Raji\\/Burkitt lymphoma and a squamous cell carcinoma of the tongue. A 10- to 20-fold increase in resistance relative to the parental line was achieved in 3-4 months of continuous selection pressure. At this time,

Emil Frei; Carol A. Cucchi; Andre Rosowsky; Ramana Tantravahi; Samuel Bernal; Thomas J. Ervin; Ruth M. Ruprecht; William A. Haseltine



Isolation of two chloroethylnitrosourea-sensitive Chinese hamster cell lines  

SciTech Connect

1-((4-Amino-2-methylpyrimidin-5-yl)methyl)-3-(2-chloroethyl)-3- nitrosourea hydrochloride (ACNU), a cancer chemotherapeutic bifunctional alkylating agent, causes chloroethylation of DNA and subsequent DNA strand cross-linking through an ethylene bridge. We isolated and characterized two ACNU-sensitive mutants from mutagenized Chinese hamster ovary cells and found them to be new drug-sensitive recessive Chinese hamster mutants. Both mutants were sensitive to various monofunctional alkylating agents in a way similar to that of the parental cell lines CHO9. One mutant (UVS1) was cross-sensitive to UV and complemented the UV sensitivity of all Chinese hamster cell lines of 7 established complementation groups. Since UV-induced unscheduled DNA synthesis was very low, a new locus related to excision repair is thought to be defective in this cell line. Another ACNU-sensitive mutant, CNU1, was slightly more sensitive to UV than the parent cell line. CNU1 was cross-sensitive to 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea and slightly more sensitive to mitomycin C. No increased accumulation of ACNU and a low level of UV-induced unscheduled DNA synthesis in this cell as compared with the parental cell line suggest that there is abnormality in a repair response of this mutant cell to some types of DNA cross-links.

Hata, H.; Numata, M.; Tohda, H.; Yasui, A.; Oikawa, A. (Tohoku Univ., Sendai (Japan))



Baculovirus studies in new, indigenous lepidopteran cell lines.  


Eight lepidopteran cell lines were established recently and their susceptibility to different insect viruses was studied. Two Spodoptera litura cell lines from the larval and pupal ovaries, were found highly susceptible to S. litura nuclear polyhedrosis virus (SLNPV, 5-6 x 10(6) NPV/ml). The Helicoverpa armigera cell line from the embryonic tissue was highly susceptible to H. armigera NPV (HaNPV, 6.3 x 10(6) NPV/ml). These in vitro grown SLNPV and HaNPV caused 100% mortality to respective 2nd instar larvae. The susceptibility of the cryo-preserved cell lines to respective baculoviruses (SLNPV/HaNPV) was studied and no significant difference in their susceptibility status was observed. The cultures could grow as suspension culture on shakers and may find application for in vitro production of wild type/recombinant baculoviruses as bio-insecticides. S. litura and Bombyx mori cell lines from larval ovaries, were highly susceptible to Autographa californica NPV (5.5 x 10(6) NPV/ml) and Bombyx mori NPV (BmNPV, 6.1 x 10(6) NPV/ml) respectively. These cell lines may find application in baculovirus expression vector studies for the production of recombinant proteins, useful in the development of diagnostic kits or as vaccines. PMID:12561971

Pant, U; Sudeep, A B; Athawale, S S; Vipat, V C



Metronidazole Decreases Viability of DLD-1 Colorectal Cancer Cell Line.  


Abstract The aim of our study was to evaluate the impact of metronidazole (MTZ) on DLD-1 colorectal cancer cell (CRC) line. Toxicity of MTZ was determined by MTT test. Cells were incubated with MTZ used in different concentrations for 24, 48, and 72 hours. The effect of MTZ on DNA synthesis was measured as [3H]-thymidine incorporation. The morphological changes in human DLD-1 cell line were defined by transmission electron microscope OPTON 900. The influence of MTZ on the apoptosis of DLD-1 cell lines was detected by flow cytometry and fluorescence microscopy, while cell concentration, volume, and diameter were displayed by Scepter Cell Counter from Millipore. Our results show that cell viability was diminished in all experimental groups in comparison with the control, and the differences were statistically significant. We did not find any significant differences in [3H]-thymidine incorporation in all experimental groups and times of observation. Cytofluorimetric assays demonstrated a statistically significant increase of apoptotic rate in MTZ concentrations 10 and 50??g/mL after 24 hours; 0.1, 10, 50, and 250??g/mL after 48 hours; and in all concentrations after 72 hours compared with control groups. In the ultrastructural studies, necrotic or apoptotic cells were occasionally seen. In conclusion, MTZ affects human CRC cell line viability. The reduction of cell viability was consistent with the apoptotic test. PMID:23777253

Sadowska, Anna; Kr?towski, Rafa?; Szynaka, Beata; Cechowska-Pasko, Marzanna; Car, Halina



Variation in Hematopoietic Potential of Induced Pluripotent Stem Cell Lines  

Microsoft Academic Search

Induced pluripotent stem (iPS) cells were originally generated from somatic cells by ectopic expression of four transcription\\u000a factor genes: Oct3\\/4, Sox2, Klf4 and c-Myc. Currently, iPS cell lines differ in tissue origin, the combination of factors used to construct them, the method of gene\\u000a delivery and expression of pluripotency markers. Thus to evaluate iPS cells for haematotherapy, the hematopoietic potential

Kasem Kulkeaw; Yuka Horio; Chiyo Mizuochi; Minetaro Ogawa; Daisuke Sugiyama



Fibronectin Synthesized by a Human Hepatoma Cell Line1  

Microsoft Academic Search

Fibronectin is a family of immunologically similar glycoproteins which mediate a variety of cell-cell and cell-substratum interac tions. It is a constituent of the extracellular matrix of connective tissue and circulates in plasma. When suspension and adherent cultures of a human hepatoma cell line (SK-HEP-1) were incu bated in serum-free medium, the resulting conditioned medium contained material which was specifically

James E. Glasgow; Robert W. Colman


Fibronectin synthesized by a human hepatoma cell line  

Microsoft Academic Search

Fibronectin is a family of immunologically similar glycoproteins which mediate a variety of cell-cell and cell-substratum interactions. It is a constituent of the extracellular matrix of connective tissue and circulates in plasma. When suspension and adherent cultures of a human hepatoma cell line (SK-HEP-1) were incubated in serum-free medium, the resulting conditioned medium contained material which was specifically immunoprecipitated by

J. E. Glasgow; R. W. Colman



Cancer Stem Cell-Like Side Population Cells in Clear Cell Renal Cell Carcinoma Cell Line 769P  

PubMed Central

Although cancers are widely considered to be maintained by stem cells, the existence of stem cells in renal cell carcinoma (RCC) has seldom been reported, in part due to the lack of unique surface markers. We here identified cancer stem cell-like cells with side population (SP) phenotype in five human RCC cell lines. Flow cytometry analysis revealed that 769P, a human clear cell RCC cell line, contained the largest amount of SP cells as compared with other four cell lines. These 769P SP cells possessed characteristics of proliferation, self-renewal, and differentiation, as well as strong resistance to chemotherapy and radiotherapy that were possibly related to the ABCB1 transporter. In vivo experiments with serial tumor transplantation in mice also showed that 769P SP cells formed tumors in NOD/SCID mice. Taken together, these results indicate that 769P SP cells have the properties of cancer stem cells, which may play important roles in tumorigenesis and therapy-resistance of RCC.

Yao, Zhi Jun; Chen, Xu; Guo, Sheng Jie; Mao, Xiao Peng; Wang, Dao Hu; Chen, Jun Xing; Qiu, Shao Peng



Comparative analysis of cell death induction by Taurolidine in different malignant human cancer cell lines  

Microsoft Academic Search

BACKGROUND: Taurolidine (TRD) represents an anti-infective substance with anti-neoplastic activity in many malignant cell lines. So far, the knowledge about the cell death inducing mechanisms and pathways activated by TRD is limited. The aim of this study was therefore, to perform a comparative analysis of cell death induction by TRD simultaneously in different malignant cell lines. MATERIALS AND METHODS: Five

Ansgar M Chromik; Adrien Daigeler; Daniel Bulut; Annegret Flier; Christina May; Kamran Harati; Jan Roschinsky; Dominique Sülberg; Peter R Ritter; Ulrich Mittelkötter; Stephan A Hahn; Waldemar Uhl



Transcription profiles of non-immortalized breast cancer cell lines  

PubMed Central

Background Searches for differentially expressed genes in tumours have made extensive use of array technology. Most samples have been obtained from tumour biopsies or from established tumour-derived cell lines. Here we compare cultures of non-immortalized breast cancer cells, normal non-immortalized breast cells and immortalized normal and breast cancer cells to identify which elements of a defined set of well-known cancer-related genes are differentially expressed. Methods Cultures of cells from pleural effusions or ascitic fluids from breast cancer patients (MSSMs) were used in addition to commercially-available normal breast epithelial cells (HMECs), established breast cancer cell lines (T-est) and established normal breast cells (N-est). The Atlas Human Cancer 1.2 cDNA expression array was employed. The data obtained were analysed using widely-available statistical and clustering software and further validated through real-time PCR. Results According to Significance Analysis of Microarray (SAM) and AtlasImage software, 48 genes differed at least 2-fold in adjusted intensities between HMECs and MSSMs (p < 0.01). Some of these genes have already been directly linked with breast cancer, metastasis and malignant progression, whilst others encode receptors linked to signal transduction pathways or are otherwise related to cell proliferation. Fifty genes showed at least a 2.5-fold difference between MSSMs and T-est cells according to AtlasImage, 2-fold according to SAM. Most of these classified as genes related to metabolism and cell communication. Conclusion The expression profiles of 1176 genes were determined in finite life-span cultures of metastatic breast cancer cells and of normal breast cells. Significant differences were detected between the finite life-span breast cancer cell cultures and the established breast cancer cell lines. These data suggest caution in extrapolating information from established lines for application to clinical cancer research.

Fernandez-Cobo, Mariana; Holland, James F; Pogo, Beatriz GT



Novel cell lines established from pediatric brain tumors  

PubMed Central

The paucity of cell culture models for childhood brain tumors prompted us to establish pediatric cell lines for use in biological experiments and preclinical developmental therapeutic studies. Three cell lines were established, CHLA-200 (GBM), CHLA-259 (anaplastic medulloblastoma) and CHLA-266 (atypical teratoid rhabdoid tumor, AT/RT). Consistent with an AT/RT origin, CHLA-266 lacked INI1 expression and had monosomy 22. All lines had unique DNA short tandem repeat “fingerprints” matching that of the patient’s tumor tissue and were adherent on tissue culture plastic, but differed in morphology and doubling times. CHLA-200 had a silent mutation in TP53. CHLA-259 and CHLA-266 had wild-type TP53. All three lines were relatively resistant to multiple drugs when compared to the DAOY medulloblastoma cell line, using the DIMSCAN fluorescence digital image microscopy cytotoxicity assay. RNA expression of MYC and MYCN were quantified using RT-PCR (Taqman). CHLA-200 expressed MYC, DAOY and CHLA-259 expressed MYCN, and CHLA-266 expressed both MYCN and MYC. CHLA-200 was only tumorigenic subcutaneously, but CHLA-259 and CHLA-266 were tumorigenic both subcutaneously and in brains of NOD/SCID mice. Immunohistochemistry of the xenografts revealed GFAP staining in CHLA-200 and PGP 9.5 staining in CHLA-259 and CHLA-266 tumors. As expected, INI1 expression was lacking in CHLA-266 (AT/RT). These three new cell lines will provide useful models for research of pediatric brain tumors.

Xu, Jingying; Erdreich-Epstein, Anat; Gonzalez-Gomez, Ignacio; Melendez, Elizabeth Y.; Smbatyan, Goar; Moats, Rex A.; Rosol, Michael; Biegel, Jaclyn A.



Stem-like Cells in Bladder Cancer Cell Lines with Differential Sensitivity to Cisplatin  

PubMed Central

Background Recurrence is a common problem in bladder cancer; this has been attributed to cancer stem cells. In this study, we characterized potential cancer stem cell populations isolated from three cell lines that demonstrate different responses to cisplatin. Materials and Methods The ALDEFLUOR® assay was used to isolate cells from TCCSUP, T24, and 5637 cell lines, and these cells were evaluated for their ability to form colonies, differentiate, migrate and invade. Results The cell lines demonstrate a spectrum of aldehyde dehydrogenase high (ALDHHigh)populations that correlate with resistance to cisplatin. In the two resistant cell lines, T24 and 5637, the ALDHHigh cells demonstrate increased colony formation, migration, invasion, and ability to differentiate. The resistant T24 and 5637 cell lines may serve as models to investigate alternative therapies for bladder cancer.

Sarachine Falso, Miranda J.; Buchholz, Bruce A.; deVere White, Ralph W.



[Effect of NKG2D in eliminating hematological malignant cell lines by natural killer cells].  


The aim of this study was to clarify whether NKG2D plays an activating role in eliminating hematological malignant cells lines by natural killer (NK) cells. Several hematological malignant cell lines (K562, NB4, Kasumi-1 THP-1, MV-4-11, MOLT-4, Jurkat, RS4; 11, Raji) were used as target cells. The expression levels of major histocompatibility complex class I (MHC I)-related molecules A/B (MICA, MICB), whose corresponding ligand was NKG2D, were detected in target cells by flow cytometry. Firstly, the target cell lines were co-incubated with carboxyfluorescein succinimidyl ester (CFSE) for 30 min. In the meanwhile, NK92MI, a kind of NK cell line, was co-incubated respectively with isotype control antibody or blocking antibody, the latter could block NKG2D specifically. Then, NK92MI cells were co-cultured with different target cell lines. After incubation for 2 h, the apoptotic ratio of each target cell line was detected by flow cytometry. The results demonstrated that there was a significant reduction of the apoptotic ratio in Kasumi-1, an acute myeloid leukemia cell line, when NK92MI cells were incubated with NKG2D blocking antibody previously. In contrast, the apoptotic ratio of other cell lines varied minimally. It is concluded that NKG2D can activate NK cells through inducing cytotoxicity to certain target cells. PMID:22541085

Wang, Wei; Gao, Li; Ma, Yi-Gai



76 FR 16609 - Proposed Information Collection; Comment Request; Identification of Human Cell Lines Project  

Federal Register 2010, 2011, 2012, 2013

...Comment Request; Identification of Human Cell Lines Project AGENCY: National Institute...repeat (STR) profiling up to 1500 human cell line samples as part of the Identification of Human Cell Lines Project. All data and...



Clonal derivation and characterization of human embryonic stem cell lines.  


Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived cell line AS034.1, respectively. The hESC line SA002 was recently reported to have an abnormal karyotype (trisomy 13), but within this population of cells we observed rare individual cells with an apparent normal karyotype. At a cloning efficiency of 5%, we established 33 subclones from SA002, out of which one had a diploid karyotype and this subline was designated SA002.5. From AS034.1 we established one reclone designated AS034.1.1 at a cloning efficiency of 0.1%. These two novel sublines express cell surface markers indicative of undifferentiated hESC (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), Oct-4, alkaline phosphatase, and they display high telomerase activity. In addition, the cells are pluripotent and form derivatives of all three embryonic germ layers in vitro as well as in vivo. These results, together with the clonal character of SA002.5 and AS034.1.1 make these homogenous cell populations very useful for hESC based applications in drug development and toxicity testing. In addition, the combination of the parental trisomic hESC line SA002 and the diploid subclone SA002.5 provides a unique experimental system to study the molecular mechanisms underlying the pathologies associated with trisomy 13. PMID:16324761

Heins, Nico; Lindahl, Anders; Karlsson, Ulrika; Rehnström, Marie; Caisander, Gunilla; Emanuelsson, Katarina; Hanson, Charles; Semb, Henrik; Björquist, Petter; Sartipy, Peter; Hyllner, Johan




EPA Science Inventory

We have measured the levels of amplification of oncogenes and tumor marker genes or other genes of interest in nine human lung tumor cell lines in comparison to normal human bronchial epithelial cells or normal blood lymphocytes to test the hypothesis that aberrant amplification ...


In vitro Rb-1 gene transfer to retinoblastoma cell lines.  

National Technical Information Service (NTIS)

After transfection of Rb-vector to packaging cell line (CRIP) by Ca-P precipitation method, we could select nineteen colonies of G-418 resistant clone by ring cloning. Each colony was transduced to NIH3T3 cells to select the one which produces high titer ...

S. W. Choi Y. H. Ham M. H. Kim



Isolation of a Primate Embryonic Stem Cell Line  

Microsoft Academic Search

Embryonic stem cells have the ability to remain undifferentiated and proliferate indefinitely in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. Here we report the derivation of a cloned cell line (R278.5) from a rhesus monkey blastocyst that remains undifferentiated in continuous passage for >1 year, maintains a normal XY karyotype, and expresses

James A. Thomson; Jennifer Kalishman; Thaddeus G. Golos; Maureen Durning; Charles P. Harris; Robert A. Becker; John P. Hearn



Derivation of human embryonic stem cell lines after blastocyst microsurgery.  


Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of the blastocyst. Because of their ability to differentiate into a variety of cell types, human embryonic stem cells (hESCs) provide an unlimited source of cells for clinical medicine and have begun to be used in clinical trials. Presently, although several hundred hESC lines are available in the word, only few have been widely used in basic and applied research. More and more hESC lines with differing genetic backgrounds are required for establishing a bank of hESCs. Here, we report the first Canadian hESC lines to be generated from cryopreserved embryos and we discuss how we navigated through the Canadian regulatory process. The cryopreserved human zygotes used in this study were cultured to the blastocyst stage, and used to isolate ICM via microsurgery. Unlike previous microsurgery methods, which use specialized glass or steel needles, our method conveniently uses syringe needles for the isolation of ICM and subsequent hESC lines. ICM were cultured on MEF feeders in medium containing FBS or serum replacer (SR). Resulting outgrowths were isolated, cut into several cell clumps, and transferred onto fresh feeders. After more than 30 passages, the two hESC lines established using this method exhibited normal morphology, karyotype, and growth rate. Moreover, they stained positively for a variety of pluripotency markers and could be differentiated both in vitro and in vivo. Both cell lines could be maintained under a variety of culture conditions, including xeno-free conditions we have previously described. We suggest that this microsurgical approach may be conducive to deriving xeno-free hESC lines when outgrown on xeno-free human foreskin fibroblast feeders. PMID:20555390

Meng, Guoliang; Liu, Shiying; Li, Xiangyun; Krawetz, Roman; Rancourt, Derrick E



Solid Oxide Fuel Cell Systems PVL Line  

SciTech Connect

In July 2010, Stark State College (SSC), received Grant DE-EE0003229 from the U.S. Department of Energy (DOE), Golden Field Office, for the development of the electrical and control systems, and mechanical commissioning of a unique 20kW scale high-pressure, high temperature, natural gas fueled Stack Block Test System (SBTS). SSC worked closely with subcontractor, Rolls-Royce Fuel Cell Systems (US) Inc. (RRFCS) over a 13 month period to successfully complete the project activities. This system will be utilized by RRFCS for pre-commercial technology development and training of SSC student interns. In the longer term, when RRFCS is producing commercial products, SSC will utilize the equipment for workforce training. In addition to DOE Hydrogen, Fuel Cells, and Infrastructure Technologies program funding, RRFCS internal funds, funds from the state of Ohio, and funding from the DOE Solid State Energy Conversion Alliance (SECA) program have been utilized to design, develop and commission this equipment. Construction of the SBTS (mechanical components) was performed under a Grant from the State of Ohio through Ohio's Third Frontier program (Grant TECH 08-053). This Ohio program supported development of a system that uses natural gas as a fuel. Funding was provided under the Department of Energy (DOE) Solid-state Energy Conversion Alliance (SECA) program for modifications required to test on coal synthesis gas. The subject DOE program provided funding for the electrical build, control system development and mechanical commissioning. Performance testing, which includes electrical commissioning, was subsequently performed under the DOE SECA program. Rolls-Royce Fuel Cell Systems is developing a megawatt-scale solid oxide fuel cell (SOFC) stationary power generation system. This system, based on RRFCS proprietary technology, is fueled with natural gas, and operates at elevated pressure. A critical success factor for development of the full scale system is the capability to test fuel cell components at a scale and under conditions that can be accurately extrapolated to full system performance. This requires specially designed equipment that replicates the pressure (up to 6.5 bara), temperature (about 910 C), anode and cathode gas compositions, flows and power generation density of the full scale design. The SBTS fuel cell anode gas is produced through the reaction of pipeline natural gas with a mixture of steam, CO2, and O2 in a catalytic partial oxidation (CPOX) reactor. Production of the fuel cell anode gas in this manner provides the capability to test a fuel cell with varying anode gas compositions ranging from traditional reformed natural gas to a coal-syngas surrogate fuel. Stark State College and RRFCS have a history of collaboration. This is based upon SSCAs commitment to provide students with skills for advanced energy industries, and RRFCS need for a workforce that is skilled in high temperature fuel cell development and testing. A key to this approach is the access of students to unique SOFC test and evaluation equipment. This equipment is designed and developed by RRFCS, with the participation of SSC interns. In the near-term, the equipment will be used by RRFCS for technology development. When this stage is completed, and RRFCS has moved to commercial products, SSC will utilize this equipment for workforce training. The RRFCS fuel cell design is based upon a unique ceramic substrate architecture in which a porous, flat substrate (tube) provides the support structure for a network of solid oxide fuel cells that are electrically connected in series. These tubes are grouped into a {approx}350-tube repeat configuration, called a stack/block. Stack/block testing, performed at system conditions, provides data that can be confidently scaled to full scale performance. This is the basis for the specially designed and developed test equipment that is required for advancing and accelerating the RRFCS SOFC power system development program. All contract DE-EE0003229 objectives were achieved and deliverables completed during the peri

Susan Shearer - Stark State College; Gregory Rush - Rolls-Royce Fuel Cell Systems



Female Sex Bias in Human Embryonic Stem Cell Lines  

PubMed Central

The factors limiting the rather inefficient derivation of human embryonic stem cells (HESCs) are not fully understood. The aim of this study was to analyze the sex ratio in our 42 preimplantation genetic diagnosis (PGD)-HESC lines, in an attempt to verify its affect on the establishment of HESC lines. The ratio between male and female PGD-derived cell lines was compared. We found a significant increase in female cell lines (76%). This finding was further confirmed by a meta-analysis for combining the results of all PGD-derived HESC lines published to date (148) and all normal karyotyped HESC lines derived from spare in vitro fertilization embryos worldwide (397). Further, gender determination of embryos demonstrated that this difference originates from the actual derivation process rather than from unequal representation of male and female embryos. It can therefore be concluded that the clear-cut tendency for female preponderance is attributed to suboptimal culture conditions rather than from a true gender imbalance in embryos used for derivation of HESC lines. We propose a mechanism in which aberrant X chromosome inactivation and/or overexpression of critical metabolic X-linked genes might explain this sex dimorphism.

Ben-Yosef, Dalit; Amit, Ami; Malcov, Mira; Frumkin, Tsvia; Ben-Yehudah, Ahmi; Eldar, Ido; Mey-Raz, Nava; Azem, Foad; Altarescu, Gheona; Renbaum, Paul; Beeri, Rachel; Varshaver, Irit; Eldar-Geva, Talia; Epsztejn-Litman, Silvina; Levy-Lahad, Ephrat



Definitive molecular cytogenetic characterization of 15 colorectal cancer cell lines.  


In defining the genetic profiles in cancer, cytogenetically aberrant cell lines derived from primary tumors are important tools for the study of carcinogenesis. Here, we present the results of a comprehensive investigation of 15 established colorectal cancer cell lines using spectral karyotyping (SKY), fluorescence in situ hybridization, and comparative genomic hybridization (CGH). Detailed karyotypic analysis by SKY on five of the lines (P53HCT116, T84, NCI-H508, NCI-H716, and SK-CO-1) is described here for the first time. The five lines with karyotypes in the diploid range and that are characterized by defects in DNA mismatch repair had a mean of 4.8 chromosomal abnormalities per line, whereas the 10 aneuploid lines exhibited complex karyotypes and a mean of 30 chromosomal abnormalities. Of the 150 clonal translocations, only eight were balanced and none were recurrent among the lines. We also reviewed the karyotypes of 345 cases of adenocarcinoma of the large intestine listed in the Mitelman Database of Chromosome Aberrations in Cancer. The types of abnormalities observed in the cell lines reflected those seen in primary tumors: there were no recurrent translocations in either tumors or cell lines; isochromosomes were the most common recurrent abnormalities; and breakpoints occurred most frequently at the centromeric/pericentromeric and telomere regions. Of the genomic imbalances detected by array CGH, 87% correlated with chromosome aberrations observed in the SKY studies. The fact that chromosome abnormalities predominantly result in copy number changes rather than specific chromosome or gene fusions suggests that this may be the major mechanism leading to carcinogenesis in colorectal cancer. PMID:19927377

Knutsen, Turid; Padilla-Nash, Hesed M; Wangsa, Danny; Barenboim-Stapleton, Linda; Camps, Jordi; McNeil, Nicole; Difilippantonio, Michael J; Ried, Thomas



The pursuit of ES cell lines of domesticated ungulates.  


In contrast to differentiated cells, embryonic stem cells (ESC) maintain an undifferentiated state, have the ability to self-renew, and exhibit pluripotency, i.e., they can give rise to most if not all somatic cell types and to the germ cells, egg and sperm. These characteristics make ES cell lines important resources for the advancement of human regenerative medicine, and, if established for domesticated ungulates, would help make possible the improvement of farm animals through their contribution to genetic engineering technology. Combining other genetic engineering technologies, such as somatic cell nuclear transfer with ESC technology may result in synergistic gains in the ability to precisely make and study genetic alterations in mammals. Unfortunately, despite significant advances in our understanding of human and mouse ESC, the derivation of ES cell lines from ungulate species has been unsuccessful. This may result from a lack of understanding of species-specific mechanisms that promote or influence cell pluripotency. Thorough molecular characterizations, including the elucidation of stem cell "marker" signaling cascade hierarchy, species-appropriate pluripotency markers, and pluripotency-associated chromatin alterations in the genomes of ungulate species, should improve the chances of developing efficient, reproducible technologies for the establishment of ES cell lines of economically important species like the pig, cow, goat, sheep and horse. PMID:18612851

Talbot, Neil C; Blomberg, Le Ann



Characterization of a spontaneously immortalized bovine trabecular meshwork cell line.  


Trabecular meshwork (TM) cells have widely been used as an in vitro model for glaucoma research. However, primary TM cells suffer the disadvantages of limited cell numbers and slow rates of proliferation. We discovered a spontaneously transformed bovine TM (BTM) cell line, BTM-28T. This cell line proliferated rapidly in low-glucose culture medium but also demonstrated contact inhibition in high-glucose culture medium. BTM-28T cells expressed key TM cell markers including ?-smooth muscle actin (?-SMA), laminin and collagen IV (col IV). Also, 100 nM dexamethasone (DEX) enhanced the formation of cross-linked actin networks (CLANs) in confluent BTM-28T cell cultures. Transforming growth factor beta 2 (TGF?2) induced the expression of fibronectin (FN), plasminogen activator inhibitor-1 (PAI-1), and connective tissue growth factor (CTGF) in our cell cultures. This cell line will be helpful to better understand the aqueous humor outflow pathway as related to the pathophysiology of glaucoma. PMID:23116564

Mao, Weiming; Liu, Yang; Mody, Avani; Montecchi-Palmer, Michela; Wordinger, Robert J; Clark, Abbot F



Embryonic germ cell lines and their derivation from mouse primordial germ cells.  


When primordial germ cells of the mouse are cultured on feeder layers with the addition of the polypeptide signalling molecules leukaemia inhibitory factor, Steel factor and basic fibroblast growth factor they give rise to cells that resemble undifferentiated blastocyst-derived embryonic stem cells. These primordial germ cell-derived embryonic germ cells (EG cells) can be induced to differentiate extensively in culture and also form teratocarcinomas when injected into nude mice. Additionally, they contribute to chimeras when injected into host blastocysts. We have derived multiple EG cell lines from 8.5 days post coitum (dpc) embryos of C57BL/6 inbred mice. Four independent EG cell lines with normal male karyotypes have formed chimeras (up to 70% coat colour chimerism) when injected into BALB/c host blastocysts. Chimeric mice from all four cell lines are fertile, but only those from one line have transmitted coat colour markers through the germline. Studies have also been carried out to determine whether gonadal primordial germ cells can give rise to pluripotent EG cells. Germ cells from gonads of 15.5 dpc C57BL/6 embryos and newborn mice failed to produce EG cell lines. EG cell lines capable of forming teratocarcinomas and coat colour chimeras have been established from primordial germ cells of 12.5 dpc genital ridges. We are currently testing the genomic imprinting status of the insulin-like growth factor type 2 receptor gene (Igf2r) in our different EG cell lines. PMID:7835148

Labosky, P A; Barlow, D P; Hogan, B L



The Organelle Proteome of the DT40 Lymphocyte Cell Line*S?  

PubMed Central

A major challenge in eukaryotic cell biology is to understand the roles of individual proteins and the subcellular compartments in which they reside. Here, we use the localization of organelle proteins by isotope tagging technique to complete the first proteomic analysis of the major organelles of the DT40 lymphocyte cell line. This cell line is emerging as an important research tool because of the ease with which gene knockouts can be generated. We identify 1090 proteins through the analysis of preparations enriched for integral membrane or soluble and peripherally associated proteins and localize 223 proteins to the endoplasmic reticulum, Golgi, lysosome, mitochondrion, or plasma membrane by matching their density gradient distributions to those of known organelle residents. A striking finding is that within the secretory and endocytic pathway a high proportion of proteins are not uniquely localized to a single organelle, emphasizing the dynamic steady-state nature of intracellular compartments in eukaryotic cells.

Hall, Stephanie L.; Hester, Svenja; Griffin, Julian L.; Lilley, Kathryn S.; Jackson, Antony P.



Omeprazole inhibits growth of cancer cell line of colonic origin.  


The direct effects of omeprazole on colonic cells has not been evaluated. Controversy exists regarding the potential adverse effects of omeprazole on cell proliferation. In order to mimic the in vivo situation in the patient treated with omeprazole, proliferation cell culture experiments were performed, monitoring directly the effects of gastrin and omeprazole both alone and in combination. Three colonic cancer cell lines were used, two with neuroendocrine features (NCI-H716, LCC-18) and one (DLD-1) not known to have these features. In these in vitro proliferation experiments, only the NCI-H716 colorectal cancer cell line responded to omeprazole by decreased proliferation (P < 0.05). The effect was concentration dependent shown for all doses of omeprazole used. Gastrin had a statistically significant effect on increasing proliferation in the NCS-H716 cell line alone but only at the highest concentration (10(-6) M). Omeprazole has a cytostatic effect on one of three colorectal cancer cell lines but the mechanism for this effect of omeprazole and its potential role in treatment awaits elucidation. PMID:7628278

Tobi, M; Chintalapani, S; Goo, R; Maliakkal, B; Reddy, J; Lundqvist, M; Oberg, K; Luk, G



Comparative Metabolic Flux Profiling of Melanoma Cell Lines  

PubMed Central

Metabolic rewiring is an established hallmark of cancer, but the details of this rewiring at a systems level are not well characterized. Here we acquire this insight in a melanoma cell line panel by tracking metabolic flux using isotopically labeled nutrients. Metabolic profiling and flux balance analysis were used to compare normal melanocytes to melanoma cell lines in both normoxic and hypoxic conditions. All melanoma cells exhibited the Warburg phenomenon; they used more glucose and produced more lactate than melanocytes. Other changes were observed in melanoma cells that are not described by the Warburg phenomenon. Hypoxic conditions increased fermentation of glucose to lactate in both melanocytes and melanoma cells (the Pasteur effect). However, metabolism was not strictly glycolytic, as the tricarboxylic acid (TCA) cycle was functional in all melanoma lines, even under hypoxia. Furthermore, glutamine was also a key nutrient providing a substantial anaplerotic contribution to the TCA cycle. In the WM35 melanoma line glutamine was metabolized in the “reverse” (reductive) direction in the TCA cycle, particularly under hypoxia. This reverse flux allowed the melanoma cells to synthesize fatty acids from glutamine while glucose was primarily converted to lactate. Altogether, this study, which is the first comprehensive comparative analysis of metabolism in melanoma cells, provides a foundation for targeting metabolism for therapeutic benefit in melanoma.

Scott, David A.; Richardson, Adam D.; Filipp, Fabian V.; Knutzen, Christine A.; Chiang, Gary G.; Ronai, Ze'ev A.; Osterman, Andrei L.; Smith, Jeffrey W.



Characterization of immunotoxins active against ovarian cancer cell lines.  

PubMed Central

The purpose of the present study was to develop immunotoxins directed against human ovarian carcinoma cells. Four monoclonal antibodies (260F9, 454C11, 280D11, and 245E7) were chosen because they were found to bind to various ovarian carcinoma cell lines. These antibodies were covalently linked to either Pseudomonas exotoxin (PE) or ricin A chain (RTA), and the conjugates were tested against five ovarian cancer cell lines (OVCAR-2, -3, -4, -5; A1847). The ability of the immunotoxins to inhibit both protein synthesis and colony formation was evaluated. Qualitatively similar results were obtained for both types of assays. Usually, PE conjugates were more toxic than their corresponding RTA conjugates. 454C11-PE was very toxic for all ovarian carcinoma lines, whereas 454C11-RTA had low activity. Both 260F9-PE and 260F9-RTA were active in all OVCAR cell lines but not in A1847 cells. 280D11-PE was toxic for OVCAR-4; otherwise, 280D11-PE and RTA conjugates of both 280D11 and 245E7 had little activity. Specificity of immunotoxin action was shown by competition by excess antibody, nontoxicity in nontarget cells, and inactivity of an irrelevant immunotoxin. To investigate the basis of antibody-dependent differences in activity of the various immunotoxins, antibody uptake was studied in OVCAR-2 cells, and the results indicate that antibody internalization is one important factor in the activity of immunotoxins. Images

Pirker, R; FitzGerald, D J; Hamilton, T C; Ozols, R F; Laird, W; Frankel, A E; Willingham, M C; Pastan, I



Proteoglycans Secreted by Packaging Cell Lines Inhibit Retrovirus Infection  

Microsoft Academic Search

Using a model recombinant retrovirus encoding theEscherichia coli lacZgene, we have found that medium conditioned with NIH 3T3 cells and packaging cell lines derived from NIH 3T3 cells inhibits infection. Most of the inhibitory activity was greater than 100 kDa and was sensitive to chondroitinase ABC digestion, which is consistent with the inhibitor being a chondroitin sulfate proteoglycan. Proteoglycans secreted




Derivation of human embryonic stem cell lines from parthenogenetic blastocysts  

Microsoft Academic Search

Parthenogenesis is one of the main, and most useful, methods to derive embryonic stem cells (ESCs), which may be an important source of histocompatible cells and tissues for cell therapy. Here we describe the derivation and characterization of two ESC lines (hPES-1 and hPES-2) from in vitro developed blastocysts following parthenogenetic activation of human oocytes. Typical ESC morphology was seen,

Qingyun Mai; Yang Yu; Tao Li; Liu Wang; Mei-jue Chen; Shu-zhen Huang; Canquan Zhou; Qi Zhou



An established avian fibroblast cell line without mitochondrial DNA  

Microsoft Academic Search

tAn established avian fibroblast cell line (LSCC-H32) has been found to be inherently resistant to the growth-inhibitory effect of ethidium bromide, when supplied with exogenous urdine. After long-term exposure to ethidium bromide (90 days), the cell population has been transferred to drug-free medium for 60 days, and then seeded at low cell density. Three clones have been isolated and propagated

Paul Desjardins; Jean-Marc de Muys; Réjean Morais



Biological behaviors and proteomics analysis of hybrid cell line EAhy926 and its parent cell line A549  

PubMed Central

Background It is well established that cancer cells can fuse with endothelial cells to form hybrid cells spontaneously, which facilitates cancer cells traversing the endothelial barrier to form metastases. However, up to now, little is known about the biologic characteristics of hybrid cells. Therefore, we investigate the malignant biologic behaviors and proteins expression of the hybrid cell line EAhy926 with its parent cell line A549. Methods Cell counting and flow cytometry assay were carried out to assess cell proliferation. The number of cells attached to the extracellular matrix (Matrigel) was measured by MTT assay for the adhesion ability of cells. Transwell chambers were established for detecting the ability of cell migration and invasion. Tumor xenograft test was carried out to observe tumorigenesis of the cell lines. In addition, two-dimensional electrophoresis (2-DE) and mass spectrometry were utilized to identify differentially expressed proteins between in Eahy926 cells and in A549 cells. Results The doubling time of EAhy926 cell and A549 cell proliferation was 25.32 h and 27.29 h, respectively (P > 0.1). Comparing the phase distribution of cell cycle of EAhy926 cells with that of A549 cells, the percentage of cells in G0/G1 phase, in S phase and in G2/M phase was (63.7% ± 2.65%) VS (60.0% ± 3.17%), (15.4% ± 1.52%) VS (13.8% ± 1.32%), and (20.9% ± 3.40%) VS (26.3% ± 3.17%), respectively (P > 0.05). For the ability of cell adhesion of EAhy926 cells and A549 cells, the value of OD in Eahy926 cells was significantly higher than that in A549 cells (0.3236 ± 0.0514 VS 0.2434 ± 0.0390, P < 0.004). We also found that the migration ability of Eahy926 cells was stronger than that of A549 cells (28.00 ± 2.65 VS 18.00 ± 1.00, P < 0.01), and that the invasion ability of Eahy926 cells was significantly weak than that of A549 cells (15.33 ± 0.58 VS 26.67 ± 2.52, P < 0.01). In the xenograft tumor model, expansive masses of classic tumor were found in the A549 cells group, while subcutaneous inflammatory focuses were found in the EAhy926 cells group. Besides, twenty-eight proteins were identified differentially expressed between in EAhy926 cells and in A549 cells by proteomics technologies. Conclusion As for the biological behaviors, the ability of cell proliferation in Eahy926 cells was similar to that in A549 cells, but the ability in adhesion and migration of Eahy926 cells was higher. In addition, Eahy926 cells had weaker ability in invasion and could not form tumor mass. Furthermore, there were many differently expressed proteins between hybrid cell line Eahy926 cells and A549 cells, which might partly account for some of the differences between their biological behaviors at the molecular level. These results may help to understand the processes of tumor angiogenesis, invasion and metastasis, and to search for screening method for more targets for tumor therapy in future.

Lu, Ze Jun; Ren, Ya Qiong; Wang, Guo Ping; Song, Qi; Li, Mei; Jiang, Sa Sa; Ning, Tao; Guan, Yong Song; Yang, Jin Liang; Luo, Feng



Implantation of Vascular Grafts Lined with Genetically Modified Endothelial Cells  

NASA Astrophysics Data System (ADS)

The possibility of using the vascular endothelial cell as a target for gene replacement therapy was explored. Recombinant retroviruses were used to transduce the lacZ gene into endothelial cells harvested from mongrel dogs. Prosthetic vascular grafts seeded with the genetically modified cells were implanted as carotid interposition grafts into the dogs from which the original cells were harvested. Analysis of the graft 5 weeks after implantation revealed genetically modified endothelial cells lining the luminal surface of the graft. This technology could be used in the treatment of atherosclerosis disease and the design of new drug delivery systems.

Wilson, James M.; Birinyi, Louis K.; Salomon, Robert N.; Libby, Peter; Callow, Allan D.; Mulligan, Richard C.



The cell line data base and the new catalogue: detailed information on 2650 human and animal cell lines.  


The Cell Line Data Base (CLDB), set up within the Interlab Project, is a relational database containing data on 2650 human and animal cell lines which are available in labs and cell banks all over Europe. The second edition of the catalogue, directly generated from the database, has been produced, and will be published in the first months of 1993. Furthermore, the electronic catalogue is available for IBM-compatible personal computers and the version for MacIntosh is under preparation. PMID:7763741

Manniello, A; Aresu, O; Parodi, B; Romano, P; Iannotta, B; Rondanina, G; Viegi, L; Ruzzon, T



The cell line data base and the new catalogue: Detailed information on 2650 human and animal cell lines.  


The Cell Line Data Base (CLDB), set up within the Interlab Project, is a relational database containing data on 2650 Human and animal cell lines which are available in labs and cell banks all over Europe. The second edition of the catalogue, directly generated from the database, has been produced, and will be published in the first months of 1993. Furthermore, the electronic catalogue is available for IBM-compatible personal computers and the version for MacIntosh is under preparation. PMID:22358677

Manniello, A; Aresu, O; Parodi, B; Romano, P; Iannotta, B; Rondanina, G; Viegi, L; Ruzzon, T



Establishment and characterization of five cell lines derived from human malignant gliomas  

Microsoft Academic Search

We established and characterized five cell lines derived from human malignant gliomas (four glioblastomas multiforme and one highly anaplastic astrocytoma). All cell lines exhibited tumor cell morphology and growth kinetics, and anchorage-independent growth in soft agar. Cytogenetic analysis revealed significant aneuploidy in all five cases as well as clonal chromosomal alterations unique to each cell line. No cell line was

J. T. Rutka; J. R. Giblin; D. Y. Dougherty; H. C. Liu; J. R. McCulloch; C. W. Bell; R. S. Stern; C. B. Wilson; M. L. Rosenblum



Gene expression analysis of cell death induction by Taurolidine in different malignant cell lines  

Microsoft Academic Search

BACKGROUND: The anti-infective agent Taurolidine (TRD) has been shown to have cell death inducing properties, but the mechanism of its action is largely unknown. The aim of this study was to identify potential common target genes modulated at the transcriptional level following TRD treatment in tumour cell lines originating from different cancer types. METHODS: Five different malignant cell lines (HT29,

Ansgar M Chromik; Stephan A Hahn; Adrien Daigeler; Annegret Flier; Daniel Bulut; Christina May; Kamran Harati; Jan Roschinsky; Dominique Sülberg; Dirk Weyhe; Ulrich Mittelkötter; Waldemar Uhl



Molecular cytogenetic analysis of breast cancer cell lines  

PubMed Central

The extensive chromosome rearrangements of breast carcinomas must contribute to tumour development, but have been largely intractable to classical cytogenetic banding. We report here the analysis by 24-colour karyotyping and comparative genomic hybridization (CGH) of 19 breast carcinoma cell lines and one normal breast epithelial cell line, which provide model examples of karyotype patterns and translocations present in breast carcinomas. The CGH was compared with CGH of 106 primary breast cancers. The lines varied from perfectly diploid to highly aneuploid. Translocations were very varied and over 98% were unbalanced. The most frequent in the carcinomas were 8;11 in five lines; and 8;17, 1;4 and 1;10 in four lines. The most frequently involved chromosome was 8. Several lines showed complex multiply-translocated chromosomes. The very aneuploid karyotypes appeared to fall into two groups that evolved by different routes: one that steadily lost chromosomes and at one point doubled their entire karyotype; and another that steadily gained chromosomes, together with abnormalities. All karyotypes fell within the range seen in fresh material and CGH confirmed that the lines were broadly representative of fresh tumours. The karyotypes provide a resource for the cataloguing and analysis of translocations in these tumours, accessible at © 2000 Cancer Research Campaign

Davidson, J M; Gorringe, K L; Chin, S-F; Orsetti, B; Besret, C; Courtay-Cahen, C; Roberts, I; Theillet, C; Caldas, C; Edwards, P A W



Establishment and characterisation of two novel breast cancer cell lines.  


Two novel oestrogen receptor (ER) negative breast cancer cell lines, BCa-11 (familial) and BCa-15 (sporadic) were successfully established from primary tumours. Characterisation of these cell lines showed expression of epithelial specific antigen and cytokeratins confirming their epithelial lineage. Analysis of ultrastructure and anchorage independent growth confirmed the epithelial nature and transformed phenotype of these cells. Both cell lines showed loss of pRb, Dab2 and ERalpha and elevated levels of proliferation marker Ki67. In addition, BCa-11 cells showed loss of HOXA5, tumour suppressor genes p16(INK4A) and RARbeta as well as overexpression of CyclinD1. Elevation of DNMT1 and DNMT3B transcript levels, promoter hypermethylation of RASSF1A, RARbeta2, and HOXA5 further support their neoplastic origin. In conclusion, the two ERalpha negative breast cancer cell lines established herein have certain useful characteristics that may make them valuable for understanding the mechanism of oestrogen receptor negative breast tumours and testing new drugs. PMID:17959394

Raju Bagadi, Sarangadhara Appala; Kaur, Jatinder; Ralhan, Ranju



Caffeine augments Alprazolam induced cytotoxicity in human cell lines.  


Combined effects of alprazolam (Alp), a member of benzodiazepine group of drugs and caffeine on human cell lines, HeLa and THP1 were investigated in this study. Alp mediated cytotoxicity was enhanced while caffeine was present. The cell death was confirmed by observing morphological changes, LDH assay and membrane anisotropic study. Also such combined effects induced elevated level of ROS and depletion of GSH. The mechanism of cell death induced by simultaneous treatment of Alp and caffeine was associated with the calcium-mediated activation of mu-calpain, release of lysosomal protease cathepsin B, activation of PARP and cleavage of caspase 3. Our results indicate that, Alp alone induces apoptosis in human cells but in the presence of caffeine it augments necrosis in a well-regulated pathway. Thus our observations strongly suggest that, alprazolam and caffeine together produce severe cytotoxicity in human cell lines. PMID:19490937

Saha, Biswarup; Mukherjee, Ananda; Samanta, Saheli; Saha, Piyali; Ghosh, Anup Kumar; Santra, Chitta Ranjan; Karmakar, Parimal



Effects of curcumin on stem-like cells in human esophageal squamous carcinoma cell lines  

PubMed Central

Background Many cancers contain cell subpopulations that display characteristics of stem cells. Because these cancer stem cells (CSCs) appear to provide resistance to chemo-radiation therapy, development of therapeutic agents that target CSCs is essential. Curcumin is a phytochemical agent that is currently used in clinical trials to test its effectiveness against cancer. However, the effect of curcumin on CSCs is not well established. The current study evaluated curcumin-induced cell death in six cancer cell lines derived from human esophageal squamous cell carcinomas. Moreover, these cell lines and the ones established from cells that survived curcumin treatments were characterized. Methods Cell loss was assayed after TE-1, TE-8, KY-5, KY-10, YES-1, and YES-2 cells were exposed to 20–80 ?M curcumin for 30 hrs. Cell lines surviving 40 or 60 ?M curcumin were established from these six original lines. The stem cell markers aldehyde dehydrogenase-1A1 (ALDH1A1) and CD44 as well as NF-?B were used to compare CSC-like subpopulations within and among the original lines as well as the curcumin-surviving lines. YES-2 was tested for tumorsphere-forming capabilities. Finally, the surviving lines were treated with 40 and 60 ?M curcumin to determine whether their sensitivity was different from the original lines. Results The cell loss after curcumin treatment increased in a dose-dependent manner in all cell lines. The percentage of cells remaining after 60 ?M curcumin treatment varied from 10.9% to 36.3% across the six lines. The cell lines were heterogeneous with respect to ALDH1A1, NF-?B and CD44 expression. KY-5 and YES-1 were the least sensitive and had the highest number of stem-like cells whereas TE-1 had the lowest. The curcumin-surviving lines showed a significant loss in the high staining ALDH1A1 and CD44 cell populations. Tumorspheres formed from YES-2 but were small and rare in the YES-2 surviving line. The curcumin-surviving lines showed a small but significant decrease in sensitivity to curcumin when compared with the original lines. Conclusion Our results suggest that curcumin not only eliminates cancer cells but also targets CSCs. Therefore, curcumin may be an effective compound for treating esophageal and possibly other cancers in which CSCs can cause tumor recurrence.



Toxic effects induced by curcumin in human astrocytoma cell lines.  


Abstract Objective: The objective of this study was to describe the toxicity induced by curcumin in human astrocytoma cell lines. Methods: The effects induced by curcumin, at 100?µM for 24?h, were evaluated in four astrocytoma cell lines using crystal violet assay and through the evaluation of morphological and ultrastructural changes by electron microscopy. Also, the results of vital staining with acridine orange and propidium iodide for acidic vesicles and apoptotic bodies were analyzed and the expression of the Beclin1 gene was assessed by RT-PCR. Results: The cells treated with curcumin at 100?µM induced an inhibitory concentration50 of viability with morphological changes characterized by a progressive increase in large, non-acidic vesicles devoid of cytoplasmic components and organelles, but that conserved the cell nuclei. No DNA breakage was observed. The astrocytoma cells showed no apoptosis, necrosis or autophagy. Expression of BECLIN1 was not induced (p?cells. Conclusions: Curcumin at 100?µm induced a new type of death cell in astrocytoma cell lines. PMID:23889520

Romero-Hernández, Mirna A; Eguía-Aguilar, Pilar; Perézpeña-Diazconti, Mario; Rodríguez-Leviz, Alejandra; Sadowinski-Pine, Stanislaw; Velasco-Rodríguez, Luis A; Cortés, Julio Roberto Cáceres; Arenas-Huertero, Francisco



Regulation of telomerase activity in immortal cell lines.  


Telomerase is a ribonucleoprotein whose activity has been detected in germ line cells, immortal cells, and most cancer cells. Except in stem cells, which have a low level of telomerase activity, its activity is absent from normal somatic tissues. Understanding the regulation of telomerase activity is critical for the development of potential tools for the diagnosis and treatment of cancer. Using the telomeric repeat amplification protocol, we found that immortal, telomerase-positive, pseudodiploid human cells (HT1080 and HL60 cells) sorted by flow repressed in quiescent cells. This was true whether quiescence was induced by contact inhibition (NIH 3T3 mouse cells), growth factor removal (bromodeoxyuridine-blocked mouse myoblasts), reexpression of cellular senescence (the reversibly immortalized IDH4 cells), or irreversible cell differentiation (HL60 promyelocytic leukemia cells and C2C12 mouse myoblasts). Taken together, these results indicate that telomerase is active throughout the cell in dividing, immortal cells but that its activity is repressed in cells that exit the cell cycle. This suggests that quiescent stem cells that have the potential to express telomerase may remain unaffected by potential antitelomerase cancer therapies. PMID:8649404

Holt, S E; Wright, W E; Shay, J W



Regulation of telomerase activity in immortal cell lines.  

PubMed Central

Telomerase is a ribonucleoprotein whose activity has been detected in germ line cells, immortal cells, and most cancer cells. Except in stem cells, which have a low level of telomerase activity, its activity is absent from normal somatic tissues. Understanding the regulation of telomerase activity is critical for the development of potential tools for the diagnosis and treatment of cancer. Using the telomeric repeat amplification protocol, we found that immortal, telomerase-positive, pseudodiploid human cells (HT1080 and HL60 cells) sorted by flow repressed in quiescent cells. This was true whether quiescence was induced by contact inhibition (NIH 3T3 mouse cells), growth factor removal (bromodeoxyuridine-blocked mouse myoblasts), reexpression of cellular senescence (the reversibly immortalized IDH4 cells), or irreversible cell differentiation (HL60 promyelocytic leukemia cells and C2C12 mouse myoblasts). Taken together, these results indicate that telomerase is active throughout the cell in dividing, immortal cells but that its activity is repressed in cells that exit the cell cycle. This suggests that quiescent stem cells that have the potential to express telomerase may remain unaffected by potential antitelomerase cancer therapies.

Holt, S E; Wright, W E; Shay, J W



Genetic design of an optimized packaging cell line for gene vectors transducing human B cells  

Microsoft Academic Search

Viral gene vectors often rely on packaging cell lines, which provide the necessary factors in trans for the formation of virus-like particles. Previously, we reported on a first-generation packaging cell line for gene vectors, which are based on the B-lymphotropic Epstein–Barr virus (EBV), a human ?-herpesvirus. This 293HEK-derived packaging cell line harbors a helper virus genome with a genetic modification

E Hettich; A Janz; R Zeidler; D Pich; E Hellebrand; B Weissflog; A Moosmann; W Hammerschmidt



Molecular cytogenetic analysis of 11 new breast cancer cell lines  

PubMed Central

We describe a survey of genetic changes by comparative genomic hybridization (CGH) in 11 human breast cancer cell lines recently established in our laboratory. The most common gains took place at 8q (73%), 1q (64%), 7q (64%), 3q (45%) and 7p (45%), whereas losses were most frequent at Xp (54%), 8p (45%), 18q (45%) and Xq (45%). Many of the cell lines displayed prominent, localized DNA amplifications by CGH. One-third of these loci affected breast cancer oncogenes, whose amplifications were validated with specific probes: 17q12 (two cell lines with ERBB2 amplifications), 11q13 (two with cyclin-D1), 8p11–p12 (two with FGFR1) and 10q25 (one with FGFR2). Gains and amplifications affecting 8q were the most common genetic alterations in these cell lines with the minimal, common region of involvement at 8q22–q23. No high-level MYC (at 8q24) amplifications were found in any of the cell lines. Two-thirds of the amplification sites took place at loci not associated with established oncogenes, such as 1q41–q43, 7q21–q22, 7q31, 8q23, 9p21–p23, 11p12–p14, 15q12–q14, 16q13–q21, 17q23, 20p11–p12 and 20q13. Several of these locations have not been previously reported and may harbour important genes whose amplification is selected for during cancer development. In summary, this set of breast cancer cell lines displaying prominent DNA amplifications should facilitate discovery and functional analysis of genes and signal transduction pathways contributing to breast cancer development. © 1999 Cancer Research Campaign

Forozan, F; Veldman, R; Ammerman, C A; Parsa, N Z; Kallioniemi, A; Kallioniemi, O-P; Ethier, S P



Production of Skeletal Muscle Elements by Cell Lines Derived from Neoplastic Rat Mammary Epithelial Stem Cells  

Microsoft Academic Search

Single-cell-cloned cell lines intermediate in morphology be tween the cuboidal epithelial and fully elongated myoepithelial- like cells have been isolated from the single-cell-cloned epithelial stem cell lines Rama 25 and Rama 37 originally obtained from dimethylbenz(a)anthracene-induced mammary tumors from Sprague-Dawley and Wistar-Furth rats, respectively. These are designated Rama 25-11, Rama 25-I2, Rama 25-I4 (Sprague- Dawley) and Rama 50-55, Rama 59,

Philip S. Rudland; Damien J. Dunnington; Barry Gusterson; Paul Monaghan; Christine M. Hughes


Mediators from Cloned T Helper Cell Lines Affect Immunoglobulin Expression by B Cells  

Microsoft Academic Search

When cloned T helper cells encounter antigen presented by I-A-compatible macrophages, soluble mediators are produced that affect the differentiation and activation of normal B lymphocytes and cell lines of the B lineage. Exposure to such T cell culture supernatants causes two effects in the murine 70Z\\/3 cell line, which represents a pre-B stage of differentiation. These cells begin to synthesize

Christopher J. Paige; Max H. Schreier; Charles L. Sidman



Mediators from cloned t helper cell lines affect immunoglobulin expression by b cells  

Microsoft Academic Search

When cloned T helper cells encounter antigen presented by I-A-compatible macrophages, soluble mediators are produced that affect the differentiation and activation of normal B lymphocytes and cell lines of the B lineage. Exposure to such T cell culture supernatants causes two effects in the murine 70Z\\/3 cell line, which represents a pre-B stage of differentiation. These cells begin to synthesize

C. J. Paige; M H Schreier; C L Sidman



Generation of a human urinary bladder smooth muscle cell line.  


We report a cell line (hBSM) established from human urinary bladder wall smooth muscle that maintains most of the phenotypic characteristics of smooth muscle cells. Cells were dissociated from the muscular layer with collagenase (1 mg/ml) and collected and grown in M199 supplemented with 10% fetal calf serum and 1% antibiotic-antimycotic. Primary cultures were grown for 2 d and small colonies were isolated by placing glass rings around the colonies. These colonies were picked up with a fine-tipped Pasteur pipette and subcultured. This procedure was repeated several times until a culture with a uniform stable morphology was obtained. hBSM cells are elongated with tapered ends, and in high density cultures, they form swirls of cells arranged in parallel. These cells have a doubling time of approximately 72 h. Western blotting and immunofluorescence microscopy revealed stable expression of smooth muscle-specific proteins, including myosin isoforms (N-terminal isoforms SM-A/B and C-terminal isoforms SM1/2), SM22, ?-smooth muscle actin, h-caldesmon, Ca(2+)-dependent myosin light chain kinase, and protein kinase G. These cells contract upon exposure to 10 ?M bethanechol and this contraction is reversible by washing away the drug. Karyotyping showed tetraploidy with a modal chromosome number of 87, with multiple rearrangements. To our knowledge, the hBSM cell line is the first human cell line established from bladder wall smooth muscle that expresses both N- and C-terminal smooth muscle myosin isoforms. This cell line will provide a valuable tool for studying transcriptional regulation of smooth muscle myosin isoforms and effects of drugs on cellular function. PMID:22259013

Zheng, Yongmu; Chang, Shaohua; Boopathi, Ettickan; Burkett, Sandra; John, Mary; Malkowicz, S Bruce; Chacko, Samuel



Derivation of ductlike cell lines from a transplantable acinar cell carcinoma of the rat pancreas.  

PubMed Central

Two cell lines were derived from a transplantable acinar cell carcinoma that had been established from a primary carcinoma of the pancreas in an azaserine-treated Lewis rat. The cultured tumor cells initially produced amylase, but production of exocrine enzymes ceased after 1-2 weeks in culture. The cultured cells were tumorigenic in Lewis rats, and one line produced solid tumors composed of ductlike structures surrounded by dense fibrous tissue. The second cell line produced partially solid and partially cystic tumors with a mixed phenotype of squamous, mucinous, and glandular areas when it grew in vivo following regrafting. Both cell lines lost structural and immunohistochemical acinar cell markers while acquiring duct cell markers during culture and regrafting. These studies provide strong support for the hypothesis that ductlike carcinomas can arise from neoplastic pancreatic acinar cells in rats. Images Figure 2 Figure 3 Figure 4 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12

Pettengill, O. S.; Faris, R. A.; Bell, R. H.; Kuhlmann, E. T.; Longnecker, D. S.



Genetic and Molecular Characterization of Uveal Melanoma Cell Lines  

PubMed Central

Summary The recent identification of frequent activating mutations in GNAQ or GNA11 in uveal melanoma provides an opportunity to better understand the pathogenesis of this melanoma subtype, and to develop rational therapeutics to target the cellular effects mediated by these mutations. Cell lines from uveal melanoma tumors are an essential tool for these types of analyses. We report the mutation status of relevant melanoma genes, expression levels of proteins of interest and DNA fingerprinting of a panel of uveal melanoma cell lines used in the research community. Significance This study represents the most comprehensive molecular analysis of uveal melanoma cell lines performed to date. The data confirms the mutually exclusive nature of GNAQ and GNA11 mutations in vitro. The lack of BRAF, NRAS, KIT, PI3K, and AKT mutations reveal GNAQ and GNA11 uveal melanoma cells to be distinct among melanoma types. The data provided is intended as a reference for investigators to select appropriate model systems and assist with authentication of uveal melanoma cell lines.

Griewank, Klaus G.; Yu, Xiaoxing; Khalili, Jahan; Sozen, M. Mert; Stempke-Hale, Katherine; Bernatchez, Chantale; Wardell, Seth; Bastian, Boris C.; Woodman, Scott E.



Spontaneously arising immortal cell line of rat retinal pigmented epithelial cells.  


A continuous cell line of rat retinal pigment epithelium (RPE), named BPEI-1, has been established and characterized. Sheets of pure RPE cells, uncontaminated by choroidal or neural retinal cell types, were isolated from eyes of 7-day-old Long Evans rats and established in primary culture. The primary RPE cells became extensively spread and grew slowly for approximately 1 month, at which time a colony of small rapidly dividing cells spontaneously appeared. Following trypsinization, most of the typical primary RPE cells did not survive and were quickly outnumbered by the smaller cells, which gave rise to a cell line that was grown continuously for several hundred generations. When growing at the maximal rate in media containing 20% FBS (doubling time 18 h), the cells were fibroblastic and nearly devoid of pigment, but were capable of morphologic transition back to a pigmented, epithelioid form when cultured under low serum conditions. Evidence that these cells originated from RPE included specific immunolabeling with antibodies to cellular retinaldehyde binding protein and cytokeratin, negative GFAP immunoreactivity, and demonstration of avid phagocytosis of isolated rod outer segments by these cells. Partial characterization of choroidal cells eliminated the latter cells as possible contaminants which could have given rise to the cell line. The BPEI-1 cell line, and other rat RPE cell lines currently being developed from pigmented normal (LE, RCS rdy+p+) and retinal dystrophic (RCS p+) rats should facilitate biochemical and molecular biological approaches to study of RPE cell function in health and disease. PMID:7679997

McLaren, M J; Sasabe, T; Li, C Y; Brown, M E; Inana, G



Hypoxic cell turnover in different solid tumor lines  

SciTech Connect

Purpose: Most solid tumors contain hypoxic cells, and the amount of tumor hypoxia has been shown to have a negative impact on the outcome of radiotherapy. The efficacy of combined modality treatments depends both on the sequence and timing of the treatments. Hypoxic cell turnover in tumors may be important for optimal scheduling of combined modality treatments, especially when hypoxic cell targeting is involved. Methods and Materials: Previously we have shown that a double bioreductive hypoxic marker assay could be used to detect changes of tumor hypoxia in relation to the tumor vasculature after carbogen and hydralazine treatments. This assay was used in the current study to establish the turnover rate of hypoxic cells in three different tumor models. The first hypoxic marker, pimonidazole, was administered at variable times before tumor harvest, and the second hypoxic marker, CCI-103F, was injected at a fixed time before harvest. Hypoxic cell turnover was defined as loss of pimonidazole (first marker) relative to CCI-103F (second marker). Results: The half-life of hypoxic cell turnover was 17 h in the murine C38 colon carcinoma line, 23 h and 49 h in the human xenograft lines MEC82 and SCCNij3, respectively. Within 24 h, loss of pimonidazole-stained areas in C38 and MEC82 occurred concurrent with the appearance of pimonidazole positive cell debris in necrotic regions. In C38 and MEC82, most of the hypoxic cells had disappeared after 48 h, whereas in SCCNij3, viable cells that had been labeled with pimonidazole were still observed after 5 days. Conclusions: The present study demonstrates that the double hypoxia marker assay can be used to study changes in both the proportion of hypoxic tumor cells and their lifespan at the same time. The present study shows that large differences in hypoxic cell turnover rates may exist among tumor lines, with half-lives ranging from 17-49 h.

Ljungkvist, Anna S.E. [Department of Radiation Oncology, Radboud University Medical Center Nijmegen, Nijmegen (Netherlands) and Department of Radiation Sciences, Umeaa University, Umeaa (Sweden)]. E-mail:; Bussink, Johan [Department of Radiation Oncology, Radboud University Medical Center Nijmegen, Nijmegen (Netherlands); Kaanders, Johannes H.A.M. [Department of Radiation Oncology, Radboud University Medical Center Nijmegen, Nijmegen (Netherlands); Rijken, Paulus F.J.W. [Department of Radiation Oncology, Radboud University Medical Center Nijmegen, Nijmegen (Netherlands); Begg, Adrian C. [Division of Experimental Therapy, Netherlands Cancer Institute, Amsterdam (Netherlands); Raleigh, James A. [Department of Radiation Oncology, University of North Carolina School of Medicine, Chapel Hill, NC (United States); Kogel, Albert J. van der [Department of Radiation Oncology, Radboud University Medical Center Nijmegen, Nijmegen (Netherlands)



Ultra-wideband time-delay line inspired by composite right\\/left-handed transmission line unit cell  

Microsoft Academic Search

This paper presents a design of ultra-wideband time-delay line inspired by the composite right\\/left-handed transmission line (CRLH TL) unit cell. A rotated version of the conventional CRLH TL unit cell is used to increase the operating bandwidth. The time-delay line is optimized using computer simulation and then fabricated on a PCB for measurement. For comparison, the time-delay lines using the

J. Zhang; S. W. Cheung; T. I. Yuk



Optimized protocol for derivation of human embryonic stem cell lines.  


For the past 12 years, the biology and applications of human embryonic stem cells (hESCs) have received great attention from the scientific community. Derivatives of the first hESC line obtained by J. Thomson's group (Science 282(5391):1145-1147, 1998) have been used in clinical trials in patients with spinal cord injury, and other hESC lines have now been used to generate cells for use in treating blindness (Lancet 379(9817):713-720, 2012). In addition to the classical protocol based on mouse or human feeder layers using open culture methods (In Vitro Cellular & Developmental Biology - Animal 46(3-4):386-394, 2010; Stem Cells 23(9):1221-1227, 2005; Nature Biotechnology 24(2):185-187, 2006; Human Reproduction 21(2):503-511, 2006; Human Reproduction 20(8):2201-2206, 2005; Fertility and Sterility 83(5):1517-1529, 2005), novel hESC lines have been derived xeno-free (without using animal derived reagents) (PLoS One 5 (4):1024-1026, 2010), feeder-free (without supporting cell monolayers) (Lancet 365(9471):1601-1603, 2005), in microdrops under oil (In Vitro Cellular & Developmental Biology - Animal 46(3-4):236-41, 2010) and in suspension with ROCK inhibitor (Nature Biotechnology 28(4):361-4, 2010). Regardless of the culture system, successful hESC derivation usually requires optimization of embryo culture, the careful and timely isolation of its inner cell mass (ICM), and precise culture conditions up to the establishment of pluripotent cell growth during hESC line derivation. Herein we address the crucial steps of the hESC line derivation protocol, and provide tips to apply quality control to each step of the procedure. PMID:22614996

Camarasa, María Vicenta; Galvez, Víctor Miguel; Brison, Daniel Roy; Bachiller, Daniel



Mechanisms of prodigiosin cytotoxicity in human neuroblastoma cell lines  

Microsoft Academic Search

Prodigiosin is a bacterial red pigment with cytotoxic properties and potential antitumor activity that has been tested against different cancerous cells. In this study we report the effect and mechanisms of action of prodigiosin against different human neuroblastoma cell lines: SH-SY5Y, LAN-1, IMR-32 (N-type) and SK-N-AS (S-type). We compare the anticancerous effect of prodigiosin with that of cisplatin at different

Roser Francisco; Ricardo Pérez-Tomás; Pepita Gimènez-Bonafé; Vanessa Soto-Cerrato; Pol Giménez-Xavier; Santiago Ambrosio



Intermittent PTH Administration Converts Quiescent Lining Cells to Active Osteoblasts  

PubMed Central

Intermittent administration of parathyroid hormone (PTH) increases bone mass, at least in part, by increasing osteoblast number. One possible source of osteoblasts might be conversion of inactive lining cells to osteoblasts, and indirect evidence is consistent with this hypothesis. To better understand the possible effect of PTH on lining cell activation, a lineage tracing study was conducted using an inducible gene system. Dmp1-CreERt2 mice were crossed with ROSA26R reporter mice to render targeted mature osteoblasts and their descendents, lining cells and osteocytes, detectable by X-gal staining. Dmp1-CreERt2(+):ROSA26R mice were injected with 0.25 mg 4-OH-tamoxifen (4-OHTam) on postnatal day 3, 5, 7, 14, and 21. The animals were sacrificed on postnatal day 23, 33 or 43 (2, 12 or 22 days after the last 4-OHTam injection). On day 43, mice were challenged with a subcutaneous injection of human PTH (1–34, 80 ?g/kg) or vehicle once daily for 3 days. By 22 days after the last 4-OHTam injection, most X-gal (+) cells on the periosteal surfaces of both the calvaria and tibia were flat. Moreover, bone formation rate and collagen I(?1) mRNA expression were decreased at day 43 compared to day 23. After 3 days of PTH injections, the thickness of X-gal (+) cells increased, as did their expression of osteocalcin and collagen I(?1) mRNA. Electron microscopy revealed X-gal-associated chromagen particles in both thin cells prior to PTH administration and cuboidal cells following PTH administration. These data support the hypothesis that intermittent PTH treatment can increase osteoblast number by converting lining cells to mature osteoblasts in vivo.

Kim, Sang Wan; Pajevic, Paola Divieti; Selig, Martin; Barry, Kevin J.; Yang, Jae-Yeon; Shin, Chan Soo; Baek, Wook-Young; Kim, Jung-Eun



Conditionally immortalized cell line of inducible metanephric mesenchyme  

Microsoft Academic Search

Conditionally immortalized cell line of inducible metanephric mesenchyme.BackgroundThe mesenchymal-epithelial conversion of metanephric mesenchyme (MM) in the formation of nephronic tubules has long served as a paradigm for inductive signaling in morphogenesis. However, the mechanisms underlying this differentiation have remained an enigma due to insufficient numbers of primary mesenchymal cells that must be isolated manually from animal embryos. To overcome this

Zoia B. Levashova; Sergei Y. Plisov; Alan O. Perantoni



Decorin Suppresses Bone Metastasis in a Breast Cancer Cell Line  

Microsoft Academic Search

Decorin, the prototype of an expanding family of small leucine-rich proteoglycans, is involved in a number of cellular processes including matrix assembly, fibrillogenesis and the control of cell proliferation. In this study, we investigated the role of decorin in suppressing tumor aggressiveness and bone metastases. We used a metastatic breast cancer cell line, MDA-MB-231, to show that decorin causes marked

Kentaro Araki; Hiroki Wakabayashi; Ken Shintani; Joji Morikawa; Akihiko Matsumine; Katsuyuki Kusuzaki; Akihiro Sudo; Atsumasa Uchida



A CNS Catecholaminergic Cell Line Expresses Voltage-gated Currents  

Microsoft Academic Search

.   CATH.a is a central nervous system (CNS) catecholaminergic cell line derived from a transgenic mouse carrying the SV40 T antigen\\u000a oncogene under the transcriptional control of regulatory elements from the rat tyrosine hydroxylase gene (Suri et al., 1993).\\u000a CATH.a cells express several differentiated neuronal characteristics including medium and light chain neurofilament proteins,\\u000a synaptophysin, tyrosine hydroxylase, and dopamine ?-hydroxylase; they

M. Lazaroff; K. Dunlap; D. M. Chikaraishi



Pheochromocytoma cell lines from heterozygous neurofibromatosis knockout mice  

Microsoft Academic Search

Transplantable tumors and cell lines have been developed from pheochromocytomas arising in mice with a heterozygous knockout mutation of the neurofibromatosis gene, Nf1. Nf1 encodes a ras-GTPase-activating protein, neurofibromin, and mouse pheochromocytoma (MPC) cells in primary cultures typically show extensive spontaneous neuronal differentiation that may result from the loss of the remaining wild-type allele and defective regulation of ras signaling.

J. F. Powers; M. J. Evinger; P. Tsokas; S. Bedri; J. Alroy; M. Shahsavari; A. S. Tischler



Evolutionary dynamics of two related malignant plasma cell lines  

PubMed Central

Cancer is the consequence of sequential acquisition of mutations within somatic cells. Mutations alter the relative reproductive fitness of cells, enabling the population to evolve in time as a consequence of selection. Cancer therapy itself can select for or against specific subclones. Given the large population of tumor cells, subclones inevitably emerge and their fate will depend on the evolutionary dynamics that define the interactions between such clones. Using a combination of in vitro studies and mathematical modeling, we describe the dynamic behavior of two cell lines isolated from the same patient at different time points of disease progression and show how the two clones relate to one another. We provide evidence that the two clones coexisted at the time of initial presentation. The dominant clone presented with biopsy-proven cardiac AL amyloidosis. Initial therapy selected for the second clone that expanded leading to a change in the diagnosis to multiple myeloma. The evolutionary dynamics relating the two cell lines are discussed and a hypothesis is generated in regard to the mechanism of one of the phenotypic characteristics that is shared by these two cell lines.

Arendt, Bonnie K; Bajzer, Zeljko; Jelinek, Diane F



DNA methylation and sensitivity to antimetabolites in cancer cell lines.  


The prediction of the cellular direction of metabolic pathways toward either DNA synthesis or DNA methylation is crucial for determining the susceptibility of cancers to anti-metabolites such as fluorouracil (5-FU). We genotyped the methylenetetrahydrofolate reductase (MTHFR) gene in NCI-60 cancer cell lines, and identified the methylation status of 24 tumor suppressor genes using methylation-specific multiplex ligation-dependent probe amplification. The susceptibility of the cancer cell lines to seven antimetabolites was then determined. Cells homozygous for CC at MTHFR-A1298C were significantly more sensitive to cyclocytidine, cytarabine (AraC) and floxuridine than those with AA or AC (p=0.0215, p=0.0166, and p=0.0323, respectively), and carried more methylated tumor suppressor genes (p=0.0313). Among the 12 tumor suppressor genes which were methylated in >25% of cancer cell lines, the methylation status of TIMP3, APC and IGSF4 significantly correlated with sensitivity to pyrimidine synthesis inhibitors. In particular, cells with methylated TIMP3 had reduced mRNA levels and were significantly more sensitive to aphidicolin-glycinate, AraC and 5-FU than cells with unmethylated TIMP3. We speculate that MTHFR-A1298C homozygous CC might direct the methylation rather than the synthesis of DNA, and result in the methylation of several tumor suppressor genes such as TIMP3. These genes could be useful biological markers for predicting the efficacy of antimetabolites. PMID:18202788

Sasaki, Shin; Kobunai, Takashi; Kitayama, Joji; Nagawa, Hirokazu



Evolutionary dynamics of two related malignant plasma cell lines.  


Cancer is the consequence of sequential acquisition of mutations within somatic cells. Mutations alter the relative reproductive fitness of cells, enabling the population to evolve in time as a consequence of selection. Cancer therapy itself can select for or against specific subclones. Given the large population of tumor cells, subclones inevitably emerge and their fate will depend on the evolutionary dynamics that define the interactions between such clones. Using a combination of in vitro studies and mathematical modeling, we describe the dynamic behavior of two cell lines isolated from the same patient at different time points of disease progression and show how the two clones relate to one another. We provide evidence that the two clones coexisted at the time of initial presentation. The dominant clone presented with biopsy proven cardiac AL amyloidosis. Initial therapy selected for the second clone that expanded leading to a change in the diagnosis to multiple myeloma. The evolutionary dynamics relating the two cell lines are discussed and a hypothesis is generated in regard to the mechanism of one of the phenotypic characteristics that is shared by these two cell lines. PMID:20890105

Dingli, David; Arendt, Bonnie K; Bajzer, Zeljko; Jelinek, Diane F



Differential proteomic analysis of nuclear extracts from thyroid cell lines.  


Nuclear proteins play a major role in controlling cell functions. Differential proteomic analysis of nuclear proteins by combined 2D gel electrophoresis (2D-E) and mass spectrometry procedures can provide useful information to understand the control of cell proliferation and differentiation. To identify proteins involved in dedifferentiation, we used a differential proteomics approach by comparing nuclear extracts from the differentiated rat thyroid cell line FRTL-5 and the derived undifferentiated Ki-mol cell line, obtained by transformation with the Ki-ras oncogene. Thirteen proteins were identified as differently expressed in the nuclear compartment between the two cell lines. RT-PCR analysis performed on seven differently expressed genes showed that only in two cases the difference may be ascribable to a transcriptional mechanism. Since one of the identified proteins, namely apurinic apyrimidinic endonuclease/redox effector factor-1 (APE1/Ref-1), is suspected to play a role in thyroid tumorigenesis, we used a glutathione S-transferase (GST)-pulldown assay coupled to a 2D electrophoretic/matrix assisted laser desorption ionization-time of flight (MALDI-TOF)-mass spectrometry (MS) analysis to detect and identify its interacting partners. We show here that beta-actin directly interacted with APE1/Ref-1, as confirmed by co-immunoprecipitation assays and that this interaction was enhanced by oxidative stress on FRTL-5 cells. PMID:16431169

Salzano, Anna Maria; Paron, Igor; Pines, Alex; Bachi, Angela; Talamo, Fabio; Bivi, Nicoletta; Vascotto, Carlo; Damante, Giuseppe; Quadrifoglio, Franco; Scaloni, Andrea; Tell, Gianluca



Telomere stability genes are not mutated in osteosarcoma cell lines  

Microsoft Academic Search

Osteosarcoma (OS), the most common primary bone tumor in adolescents and young adults, is characterized by a high degree of chromosomal abnormalities. Because telomeres are important for maintaining chromosomal integrity, it is plausible that germ-line or somatic mutations in the genes responsible for stabilizing the telomere complex could contribute to OS. We performed bi-directional sequence analysis in five OS cell

Sharon A. Savage; Brian J. Stewart; Jason S. Liao; Lee J. Helman; Stephen J. Chanock



Gastric cancer cell lines induced by trichostatin A  

Microsoft Academic Search

AIM: To explore the effect of trichostatin A (TSA) on apoptosis and acetylated histone H3 levels in gastric cancer cell lines BGC-823 and SGC-7901. METHODS: The effect of TSA on growth inhibition and apoptosis was examined by MTT, fluorescence microscopy and PI single-labeled flow cytometry. The acetylated histone H3 level was detected by Western blot.

Xiao-Ming Zou; Yun-Long Li; Hao Wang; Wu Cui; Xiao-Lin Li; Song-Bin Fu; Hong-Chi Jiang



Expression of melatoninergic receptors in human placental choriocarcinoma cell lines  

Microsoft Academic Search

BACKGROUND: Melatonin crosses the placenta and enters the fetal circulation. Moreover, experimental data sug- gest a possible influence of melatonin on placental function and fetal development in humans. To date, the expression and role of melatonin receptors in human placenta choriocarcinoma cell lines and in human term placental tissues remain to be elucidated. METHODS AND RESULTS: Results from RT-PCR, western

Dave Lanoix; Rodney Ouellette; Cathy Vaillancourt




EPA Science Inventory

Diversity of arsenic metabolism in cultured human cancer cell lines. Arsenic has been known to cause a variety of malignancies in human. Pentavalent As (As 5+) is reduced to trivalent As (As3+) which is further methylated by arsenic methyltransferase(s) to monomethylarson...



EPA Science Inventory

The production of arachidonic acid metabolites by the HL60, ML3, and U937 human phagocyte cell lines were determined after incubation with interferongamma (IFNg; 500 U/ml) or vehicle for 4 days. ells were prelabeled with tritiated arachidonic acid for 4 hours, and media supernata...


Establishment and Identification of Small Cell Lung Cancer Cell Lines Having Classic and Variant Features1  

Microsoft Academic Search

Using a chemically defined medium containing hydrocortisone, insulin, transferrin, 17\\/3-estradiol and selenium, with or without serum supplementation (2.5% v\\/v), continuous cell lines can be established from 72% of all fresh biopsy specimens of small cell lung cancer (SCLC) containing tumor cells. No differences were observed in the rate of establishing cell lines from newly diag nosed untreated patients, or from

Desmond N. Carney; Adi F. Gazdar; Gerold Bepler; John G. Guccion; Paul J. Marangos; Terry W. Moody; Mark H. Zweig; John D. Minna


Cell surface proteins and glycoproteins from biologically different human colon carcinoma cell lines  

SciTech Connect

Six cultured human colon cancer cell lines possessing different biological characteristics were enzymatically radiolabeled in situ with 125I and 3H, and the labeled cell surface proteins and glycoproteins were compared. The electrophoretic patterns of labeled cell surface material suggest correlations between biological properties and cell surface proteins. Highly aggressive cell lines (as assessed by in vitro parameters) had predominant peaks of 125I-labeled proteins between molecular weights 66,000 and 92,500. The major peak of radioiodinated material from the more indolent cell lines occurred between molecular weights 31,000 and 45,000. The profile of one 125I-labeled intermediately aggressive cell line was similar to the profiles of the more aggressive lines, whereas another intermediate line exhibited a profile different from those of both indolent and aggressive lines. Electrophoresis of tritiated material indicated that essentially all of the recovered labeled glycoprotein was of relatively high molecular weight (92,000-180,000) in the indolent lines, whereas the intermediate and highly aggressive lines had patterns with significant peaks between molecular weights 45,000 and 92,500.

Marks, M.E.; Danbury, B.H.; Miller, C.A.; Brattain, M.G.



Determination of NAD+ and NADH level in a Single Cell Under H2O2 Stress by Capillary Electrophoresis  

SciTech Connect

A capillary electrophoresis (CE) method is developed to determine both NAD{sup +} and NADH levels in a single cell, based on an enzymatic cycling reaction. The detection limit can reach down to 0.2 amol NAD{sup +} and 1 amol NADH on a home-made CE-LIF setup. The method showed good reproducibility and specificity. After an intact cell was injected into the inlet of a capillary and lysed using a Tesla coil, intracellular NAD{sup +} and NADH were separated, incubated with the cycling buffer, and quantified by the amount of fluorescent product generated. NADH and NAD{sup +} levels of single cells of three cell lines and primary astrocyte culture were determined using this method. Comparing cellular NAD{sup +} and NADH levels with and without exposure to oxidative stress induced by H{sub 2}O{sub 2}, it was found that H9c2 cells respond to the stress by reducing both cellular NAD{sup +} and NADH levels, while astrocytes respond by increasing cellular NADH/NAD{sup +} ratio.

Wenjun Xi



The secretome signature of colon cancer cell lines.  


The definition of the secretome signature of a cancer cell line can be considered a potential tool to investigate tumor aggressiveness and a preclinical exploratory study required to optimize the search of cancer biomarkers. Dealing with a cell-specific secretome limits the contamination by the major components of the human serum and reduces the range of dynamic concentrations among the secreted proteins, thus favouring under-represented tissue-specific species. The aim of the present study is to characterize the secretome of two human colon carcinoma cell lines, CaCo-2 and HCT-GEO, in order to evaluate differences and similarities of two colorectal cancer model systems. In this study, we identified more than 170 protein species, 64 more expressed in the secretome of CaCo-2 cells and 54 more expressed in the secretome of HCT-GEO cells; 58 proteins were shared by the two systems. Among them, more than 50% were deemed to be secretory according to their Gene Ontology annotation and/or to their SignalP or SecretomeP scores. Such a characterization allowed corroborating the potential of a cell culture-based model in order to describe the cell-specific invasive properties and to provide a list of putative cancer biomarkers. J. Cell. Biochem. 114: 2577-2587, 2013. © 2013 Wiley Periodicals, Inc. PMID:23744648

Imperlini, Esther; Colavita, Irene; Caterino, Marianna; Mirabelli, Peppino; Pagnozzi, Daniela; Vecchio, Luigi Del; Noto, Rosa Di; Ruoppolo, Margherita; Orrù, Stefania



Transgene expression enhancement in T-lymphoma cell lines.  


In transfection protocols, the expression levels of the transgene is important to define, still is difficult to obtain in certain cell lines such as those derived from T-lymphoma cells. In this study we evaluate transgene expression kinetics in the presence and absence of two well known transcription activators such as phorbol-12-myristate13-acetate (PMA) and Ionomicin (IO). Three murine T lymphoma cell lines (LBC, EL4 and BW5147) were transfected by electroporation using green fluorescent protein (GFP) as a reporter gene and analyzed by flow cytometry. Addition of PMA/IO resulted in a significant increase of the Mean Fluorescence Intensity but not in GFP-positive cell percentages, either in transient or stable transfected LBC and EL4 cells. Remarkable, BW5147 cells showed low GFP induction with a significant increment only in stable transfected cells. Our results demonstrated that CMV promoter activity can be enhanced in transfected lymphoma cells by PMA/IO suggesting that transgene expression levels can be optimized by means of the use of transcription activators. PMID:16102518

Ruybal, Paula; Gravisaco, María José; Barcala, Virna; Escalada, Ana; Cremaschi, Graciela; Taboga, Oscar; Waldner, Claudia; Mongini, Claudia



Immortality of cell lines: challenges and advantages of establishment.  


Cellular immortality happens upon impairment of cell-cycle checkpoint pathways (p53/p16/pRb), reactivation or up-regulation of telomerase enzyme, or upregulation of some oncogenes or oncoproteins leading to a higher rate of cell division.There are also some other factors and mechanisms involved in immortalisation, which need to be discovered. Immortalisation of cells derived from different sources and establishment of immortal cell lines has proven useful in understanding the molecular pathways governing cell developmental cascades in eukaryotic, especially human, cells. After the breakthrough of achieving the immortal cells and understanding their critical importance in the field of molecular biology, intense efforts have been dedicated to establish cell lines useful for elucidating the functions of telomerase, developmental lineage of progenitors, self-renewal potency, cellular transformation, differentiation patterns and some bioprocesses, like odontogenesis. Meanwhile, discovering the exact mechanisms of immortality, a major challenge for science yet, is believed to open new gateways toward understanding and treatment of cancer in the long term. This review summarises the methods involved in establishing immortality, its advantages and the challenges still being faced in this field. PMID:23723166

Maqsood, Muhammad Irfan; Matin, Maryam M; Bahrami, Ahmad Reza; Ghasroldasht, Mohammad M



Metallothionein prevention of arsenic trioxide-induced cardiac cell death is associated with its inhibition of mitogen-activated protein kinases activation in vitro and in vivo.  


Cardiotoxicity induced by arsenic trioxide has become a serious blockade of clinical applications of this effective anticancer agent. The general mechanism responsible for arsenic cardiotoxicity has been attributed to its induction of oxidative stress. Metallothionein (MT) has been extensively proven to be a potent endogenous antioxidant that protects heart against oxidative stress-induced cardiac damage. To investigate whether and how MT protects against arsenic cardiotoxicity, MT-overexpressing H9c2 (MT-H9c2) cardiac cells and transgenic (MT-TG) mice with their corresponding controls were exposed to the clinical relevant dose of arsenic trioxide. Cardiac cell apoptosis was detected by molecular indices, including the cleavage of caspase 3 and caspase 12, Bax/Bcl2 expression ratio, CHOP expression and/or confirmed by a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Arsenic trioxide dose- and time-dependently induced cardiac cell death in H9c2 cells with a significant activation of major MAPK subfamily members such as ERK1/2, JNK and p38, but not in MT-H9c2 cells. Importantly, the protective effect of MT on arsenic trioxide-induced apoptotic cell death was completely recaptured in the heart of MT-TG with a significant prevention of MAPKs activation. These results indicate that arsenic trioxide-upregulated MAPKs might play important role in arsenic trioxide-induced apoptotic cell death in cardiac cells both in vivo and in vitro, and MT's suppression of arsenic trioxide apoptotic effect was associated with the inhibition of MAPK activation. Therefore, selective elevation of cardiac MT levels with pharmacological approaches may be a potential strategy for the prevention of arsenic cardiotoxicity. PMID:23664956

Miao, Xiao; Tang, Zefang; Wang, Yonggang; Su, Guanfang; Sun, Weixia; Wei, Wei; Li, Wei; Miao, Lining; Cai, Lu; Tan, Yi; Liu, Qiuju



Taurine regulates insulin release from pancreatic beta cell lines  

PubMed Central

Background Pancreatic ?-cells release insulin via an electrogenic response triggered by an increase in plasma glucose concentrations. The critical plasma glucose concentration has been determined to be ~3 mM, at which time both insulin and GABA are released from pancreatic ?-cells. Taurine, a ?-sulfonic acid, may be transported into cells to balance osmotic pressure. The taurine transporter (TauT) has been described in pancreatic tissue, but the function of taurine in insulin release has not been established. Uptake of taurine by pancreatic ?-cells may alter membrane potential and have an effect on ion currents. If taurine uptake does alter ?-cell current, it might have an effect on exocytosis of cytoplasmic vesicle. We wished to test the effect of taurine on regulating release of insulin from the pancreatic ?-cell. Methods Pancreatic ?-cell lines Hit-TI5 (Syrian hamster) and Rin-m (rat insulinoma) were used in these studies. Cells were grown to an 80% confluence on uncoated cover glass in RPMI media containing 10% fetal horse serum. The cells were then adapted to a serum-free, glucose free environment for 24 hours. At that time, the cells were treated with either 1 mM glucose, 1 mM taurine, 1 mM glucose + 1 mM taurine, 3 mM glucose, or 3 mM glucose + 1 mM taurine. The cells were examined by confocal microscopy for cytoplasmic levels of insulin. Results In both cell lines, 1 mM glucose had no effect on insulin levels and served as a control. Cells starved of glucose had a significant reduction (p<0.001) in the level of insulin, but this level was significantly higher than all other treatments. As expected, the 3 mM glucose treatment resulted in a statistically lower (p<0.001) insulin level than control cells. Interestingly, 1 mM taurine also resulted in a statistically lower level of insulin (p<0.001) compared to controls when either no glucose or 1 mM glucose was present. Cells treated with 1 mM taurine plus 3 mM glucose showed a level of insulin similar to that of 3 mM glucose alone. Conclusions Taurine administration can alter the electrogenic response in ?-cell lines, leading to a change in calcium homeostasis and a subsequent decrease in intracellular insulin levels. The consequence of these actions could represent a method of increasing plasma insulin levels leading to a decrease in plasma glucose levels.



9-(beta)-arabinofuranosyladenine preferentially sensitizes radioresistant squamous cell carcinoma cell lines to x-rays.  

National Technical Information Service (NTIS)

The effect of 9-(beta)-arabinofuranosyladenine (ara-A) on sensitivity to the deleterious effects of x-rays was studied in six squamous cell carcinoma cell lines. Three lines were relatively radioresistant, having D(sub 0) values of 2.31 to 2.89 Gy, and th...

D. Heaton R. Mustafi J. L. Schwartz



9-{beta}-arabinofuranosyladenine preferentially sensitizes radioresistant squamous cell carcinoma cell lines to x-rays  

SciTech Connect

The effect of 9-{beta}-arabinofuranosyladenine (ara-A) on sensitivity to the deleterious effects of x-rays was studied in six squamous cell carcinoma cell lines. Three lines were relatively radioresistant, having D{sub 0} values of 2.31 to 2.89 Gy, and the other three lines were relatively radiosensitive, having D{sub 0} values of between 1.07 and 1.45 Gy. Ara-A (50 or 500 {mu}M) was added to cultures 30 min prior to irradiation and removed 30 min after irradiation, and sensitivity was measured in terms of cell survival. The radiosensitizing effect of ara-A was very dependent on the inherent radiosensitivity of the tumor cell line. Fifty micromolar concentrations of ara-A sensitized only the two most radioresistant lines, SCC-12B.2 and JSQ-3. Five hundred micromolar concentrations of ara-A sensitized the more sensitive cell lines, SQ-20B and SQ-9G, but failed to have any effect on the radiation response of the two most sensitive cell lines, SQ-38 and SCC-61. Concentrations of ara-A as low as 10 {mu}M were equally efficient in inhibiting DNA synthesis in all six cell lines. These results suggest that the target for the radiosensitizing effect of ara-A is probably related to the factor controlling the inherent radiosensitivity of human tumor cells. Therefore, ara-A might be useful in overcoming radiation resistance in vivo.

Heaton, D. [Rush Univ. Medical Center, Chicago, IL (United States). Therapeutic Radiology; Mustafi, R. [Chicago Univ., IL (United States). Dept. of Radiation and Cellular Oncology; Schwartz, J.L. [Chicago Univ., IL (United States). Dept. of Radiation and Cellular Oncology]|[Argonne National Lab., IL (United States)



Glycosylation potential of human prostate cancer cell lines.  


Altered glycosylation is a universal feature of cancer cells and altered glycans can help cancer cells escape immune surveillance, facilitate tumor invasion, and increase malignancy. The goal of this study was to identify specific glycoenzymes, which could distinguish prostate cancer cells from normal prostatic cells. We investigated enzymatic activities and gene expression levels of key glycosyl- and sulfotransferases responsible for the assembly of O- and N-glycans in several prostatic cells. These cells included immortalized RWPE-1 cells derived from normal prostatic tissues, and prostate cancer cells derived from metastasis in bone (PC-3), brain (DU145), lymph node (LNCaP), and vertebra (VCaP). We found that all cells were capable of synthesizing complex N-glycans and O-glycans with the core 1 structure, and each cell line had characteristic biosynthetic pathways to modify these structures. The in vitro measured activities corresponded well to the mRNA levels of glycosyltransferases and sulfotransferases. Lectin and antibody binding to whole cells supported these results, which form the basis for the development of tumor cell-specific targeting strategies. PMID:22843320

Gao, Yin; Chachadi, Vishwanath B; Cheng, Pi-Wan; Brockhausen, Inka



Zebrafish kidney stromal cell lines support multilineage hematopoiesis  

PubMed Central

Studies of zebrafish hematopoiesis have been largely performed using mutagenesis approaches and retrospective analyses based upon gene expression patterns in whole embryos. We previously developed transplantation assays to test the repopulation potentials of candidate hematopoietic progenitor cells. We have been impaired, however, in determining cellular differentiation potentials by a lack of short-term functional assays. To enable more precise analyses of hematopoietic progenitor cells, we have created zebrafish kidney stromal (ZKS) cell lines. Culture of adult whole kidney marrow with ZKS cells results in the maintenance and expansion of hematopoietic precursor cells. Hematopoietic growth is dependent upon ZKS cells, and we show that ZKS cells express many growth factors and ligands previously demonstrated to be important in maintaining mammalian hematopoietic cells. In the absence of exogenous growth factors, ZKS cells maintain early hematopoietic precursors and support differentiation of lymphoid and myeloid cells. With the addition of zebrafish erythropoietin, ZKS cells also support the differentiation of erythroid precursors. These conditions have enabled the ability to ascertain more precisely the points at which hematopoietic mutants are defective. The development of robust in vitro assays now provide the means to track defined, functional outcomes for prospectively isolated blood cell subsets in the zebrafish.

Stachura, David L.; Reyes, Jason R.; Bartunek, Petr; Paw, Barry H.; Zon, Leonard I.



3-Bromopyruvate induces necrotic cell death in sensitive melanoma cell lines  

Microsoft Academic Search

Clinicians successfully utilize high uptake of radiolabeled glucose via PET scanning to localize metastases in melanoma patients. To take advantage of this altered metabolome, 3-bromopyruvate (BrPA) was used to overcome the notorious resistance of melanoma to cell death. Using four melanoma cell lines, BrPA triggered caspase independent necrosis in two lines, whilst the other two lines were resistant to killing.

J.-Z. Qin; H. Xin; B. J. Nickoloff



Trichothecene-induced cytotoxicity on human cell lines  

Microsoft Academic Search

Trichothecene cytotoxicity of type A (T-2 toxin and HT-2 toxin), type B (deoxynivalenol, DON, and nivalenol, NIV), and type\\u000a D (satratoxins G and H) compounds was determined comparatively by using eight permanent human cell lines (Hep-G2, A549, CaCo-2,\\u000a HEp-2, A204, U937, RPMI 8226, and Jurkat). Viability of cells was measured by a water-soluble tetrazolium (WST-1) reagent\\u000a cell proliferation assay assessing

Carina Nielsen; Maximilian Casteel; Andrea Didier; Richard Dietrich; Erwin Märtlbauer



Establishment and characterization of a mutagenized cell line exhibiting the 'cell-in-cell' phenotype at a high frequency.  


Cell-in-cell structures represent live cell events in which one cell internalizes another. Because formation of cell-in-cell structures is a rare event in most cell types and the event is associated with cell death, there has been limited clarification of this phenomenon, and its physiological role and molecular mechanism are yet to be precisely elucidated. In this study, we established a mutagenized cell line that exhibited cell-in-cell structures at a more than 10-fold higher frequency as compared to the parent cells. Interestingly, both engulfment and invasion were increased in the mutagenized cell line as compared with that in the parent cell line in the suspension culture condition. This finding indicates that this mutagenized cell line showed an interchangeable status in terms of its ability to form cell-in-cell structures, and the system described here could be useful for elucidation of the mechanisms regulating the formation of cell-in-cell structures, including engulfment and invasion, in a given cellular environment. Further studies using this cell line are warranted to understand the mechanism of formation and biological significance of the cell-in-cell formation. PMID:24165024

Kahyo, Tomoaki; Sugimura, Haruhiko



Radiation sensitivities of 31 human oesophageal squamous cell carcinoma cell lines  

PubMed Central

The purpose of this study was to determine the radiosensitivities of 31 human oesophageal squamous cell carcinoma cell lines with a colony-formation assay. A large variation in radiosensitivity existed among 31 cell lines. Such a large variation may partly explain the poor result of radiotherapy for this cancer. One cell line (KYSE190) demonstrated an unusual radiosensitivity. Ataxia-telangiectasia-mutated (ATM) gene in these cells had five missense mutations, and ATM protein was truncated or degraded. Inability to phosphorylate Chk2 in the irradiated KYSE190 cells suggests that the ATM protein in these cells had lost its function. The dysfunctional ATM protein may be a main cause of unusual radiosensitivity of KYSE190 cells. Because the donor of these cells was not diagnosed with ataxia telangiectasia, mutations in ATM gene might have occurred during the initiation and progression of cancer. Radiosensitive cancer developed in non-hereditary diseased patients must be a good target for radiotherapy.

Ban, Sadayuki; Michikawa, Yuichi; Ishikawa, Ken-ichi; Sagara, Masashi; Watanabe, Koji; Shimada, Yutaka; Inazawa, Johji; Imai, Takashi



Transgenic cell lines for detection of animal viruses.  

PubMed Central

Rapid diagnostic assays based on direct detection of viral antigen or nucleic acid are being used with increasing frequency in clinical virology laboratories. Virus culture, however, remains the only way to detect infectious virus and to analyze clinically relevant viral phenotypes, such as drug resistance. Growth of viruses in cell culture is labor intensive and time-consuming and requires the use of many different cell lines. Transgenic technology, together with increasing knowledge of the molecular pathways of virus replication, offers the possibility of using genetically modified cell lines to improve virus growth in cell culture and to facilitate detection of virus-infected cells. Genetically modifying cells so that they express a reporter gene only after infection with a specific virus can allow the detection of infectious virus by rapid and simple enzyme assays such as beta-galactosidase assays without the need for antibodies. Although transgenic cells have recently been successfully used for herpes simplex virus detection, much more work needs to be done to adapt this technology to other human viral pathogens such as cytomegalovirus and respiratory viruses. This review offers some strategies for applying this technology to a wide spectrum of animal viruses.

Olivo, P D



Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines  

Microsoft Academic Search

Background  Cancer stem cells (CSCs) are regarded as the cause of tumor formation and recurrence. The isolation and identification of\\u000a CSCs could help to develop novel therapeutic strategies specifically targeting CSCs.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  Human hepatoma cell lines were plated in stem cell conditioned culture system allowed for sphere forming. To evaluate the\\u000a stemness characteristics of spheres, the self-renewal, proliferation, chemoresistance, tumorigenicity of the

Lu Cao; Yanming Zhou; Beibei Zhai; Jian Liao; Wen Xu; Ruixiu Zhang; Jing Li; Yu Zhang; Lei Chen; Haihua Qian; Mengchao Wu; Zhengfeng Yin



Differences in cell proliferation in rodent and human hepatic derived cell lines exposed to ciprofibrate  

Microsoft Academic Search

Humans appear to be refractory to some effects of peroxisome proliferators including alterations in cell proliferation, whereas rodents are susceptible. In this study, differences between the human and rat response to peroxisome proliferators were evaluated using rat and human tumour liver cell lines. Rat 7777 cells were more responsive than human HepG2 cells to ciprofibrate as they exhibited a higher

Marie-Claude Clemencet; Giuliana Muzio; Antonella Trombetta; Jeffrey M. Peters; Frank J. Gonzalez; Rosa A. Canuto; Norbert Latruffe



Functionally active Epstein-Barr virus-transformed follicular dendritic cell-like cell lines  

PubMed Central

Follicular dendritic cells (FDC) are unique nonlymphoid cells found only in germinal centers. FDC can be distinguished from other accessory cells based on a characteristic set of cell surface markers. It is known that FDC are able to rescue germinal center B cells from apoptosis. To investigate the role of FDC in the process of selection and maturation of B cells during germinal center reactions, we tried to establish factor-independent immortalized FDC-like cell lines. Because freshly isolated FDC express the Epstein-Barr Virus (EBV) receptor CD21, we attempted EBV transformation on isolated FDC. After incubation of FDC-enriched cell populations with EBV, cell lines were obtained consisting of slowly duplicating very large cells. These cell lines have a fibroblast-like morphology but could be clearly distinguished from several human fibroblast cell lines by displaying a different phenotype including intercellular adhesion molecule 1, CD40, and CD75 expression. Detection of the EBV-encoded proteins latent membrane protein 1 and Epstein-Barr virus nuclear antigen 2 in our FDC-like cell lines implicated successful EBV transformation. FDC-like cells are able to bind nonautologous B cells and preserve the latter from apoptosis. The binding of B cells to FDC-like cells is dependent on adhesion via lymphocyte function-associated antigen 1/intercellular adhesion molecule 1 and closely resembles the pattern of emperipolesis as described by others. These data demonstrate that FDC can be successfully infected by EBV, and that the cell lines obtained share phenotypic and functional characteristics with freshly isolated FDC.



72 FR 34591 - Expanding Approved Stem Cell Lines in Ethically Responsible Ways  

Federal Register 2010, 2011, 2012, 2013

...Order 13435--Expanding Approved Stem Cell Lines in Ethically Responsible Ways...June 20, 2007 Expanding Approved Stem Cell Lines in Ethically Responsible Ways...respect to research on pluripotent stem cells derived by ethically...



Ganglioside GM3 Can Induce Megakaryocytoid Differentiation of Human Leukemia Cell Line K562 Cells1  

Microsoft Academic Search

The role of acidic glycosphingolipids in cell growth and differentiation was investigated using the multipotent leukemia cell line KS62. When (¡Miwas added to cell culture media, the growth of K562 cells was remarkably inhibited and the cells were shown to have megakaryocytoid morphology. Ultrastructural study demonstrated that KS62 cells treated with CM., had platelet peroxidase-positive structures, which were con sidered

Mitsuru Nakamura; Keita Kirito; Jinko Yamanoi; Tasuku Wainai; Hisao Nojiri; Masaki Saito



Cytolytic replication of echoviruses in colon cancer cell lines  

PubMed Central

Background Colorectal cancer is one of the most common cancers in the world, killing nearly 50% of patients afflicted. Though progress is being made within surgery and other complementary treatments, there is still need for new and more effective treatments. Oncolytic virotherapy, meaning that a cancer is cured by viral infection, is a promising field for finding new and improved treatments. We have investigated the oncolytic potential of several low-pathogenic echoviruses with rare clinical occurrence. Echoviruses are members of the enterovirus genus within the family Picornaviridae. Methods Six colon cancer cell lines (CaCo-2, HT29, LoVo, SW480, SW620 and T84) were infected by the human enterovirus B species echovirus 12, 15, 17, 26 and 29, and cytopathic effects as well as viral replication efficacy were investigated. Infectivity was also tested in spheroids grown from HT29 cells. Results Echovirus 12, 17, 26 and 29 replicated efficiently in almost all cell lines and were considered highly cytolytic. The infectivity of these four viruses was further evaluated in artificial tumors (spheroids), where it was found that echovirus 12, 17 and 26 easily infected the spheroids. Conclusions We have found that echovirus 12, 17 and 26 have potential as oncolytic agents against colon cancer, by comparing the cytolytic capacity of five low-pathogenic echoviruses in six colon cancer cell lines and in artificial tumors.



Identifying genes related to radiation resistance in oral squamous cell carcinoma cell lines.  


Radioresistance is one of the main determinants of treatment outcome in oral cancer, but the prediction of radioresistance is difficult. The authors aimed to establish radioresistant oral squamous cell carcinoma (OSCC) cell lines to identify genes with altered expression in response to radioresistance. To induce radioresistant cell lines, the authors treated OSCC cell lines with an accumulated dosage of 60Gy over 30 cycles of radiotherapy. They compared the results from cDNA arrays and proteomics between non-radiated and radioresistant cell lines in order to identify changes in gene expression. Western blot analysis was used to validate the results. The cDNA array revealed 265 commonly up-regulated genes and 268 commonly down-regulated genes in radioresistant cell lines, 30 of which were cancer-related genes. Proteomics identified 51 proteins with commonly altered expression in radioresistant cell lines, 18 of which were cancer-related proteins. Both the cDNA array and proteomics indicated that NM23-H1 and PA2G4 were over-expressed. Western blot analysis showed increased expression of NM23-H1, but not PA2G4, in radioresistant cell lines. The authors concluded that NM23-H1 may be a radioresistance-related gene and over-expression of NM23-H1 could serve as a biomarker to predict radioresistance in OSCC. PMID:23196067

Lee, S Y; Park, H R; Cho, N H; Choi, Y P; Rha, S Y; Park, S W; Kim, S H



Processing of proSAAS in neuroendocrine cell lines.  


ProSAAS, a recently discovered granin-like protein, potently inhibits prohormone convertase (PC)1, and might also perform additional functions. In the present study, the processing of proSAAS was compared in two neuroendocrine cell lines overexpressing this protein: the AtT-20 mouse pituitary corticotrophic line and the PC12 rat adrenal phaeochromocytoma line. The processing of proSAAS was examined by pulse-chase analysis using [(3)H]leucine, by MS, and by chromatography and radioimmunoassay. Various smaller forms of proSAAS were detected, including peptides designated as little SAAS, PEN and big LEN. Because the PC-12 cells used in the present study do not express either PC1 or PC2, the finding that these cells efficiently cleave proSAAS indicates that these cleavages do not require either enzyme. Two of the peptides identified in AtT-20 media represent novel C-terminally truncated forms of PEN. In both cell lines, the secretion of the small proSAAS-derived peptides is stimulated by secretagogues. However, long-term treatment of wild-type AtT-20 cells with two different secretagogues (8-bromo-cAMP and a phorbol ester) does not affect levels of proSAAS mRNA; this treatment significantly increases PC1 mRNA by approx. 60-80%. The lack of co-regulation of proSAAS and PC1 mRNA implies that enzyme activity can be induced without an accompanying increase in the inhibitor. In addition, the finding that the peptides are secreted via the regulated pathway is consistent with the proposal that they may function as neuropeptides. PMID:11742530

Mzhavia, Nino; Qian, Yimei; Feng, Yun; Che, Fa-Yun; Devi, Lakshmi A; Fricker, Lloyd D



Fibronectin synthesized by a human hepatoma cell line  

SciTech Connect

Fibronectin is a family of immunologically similar glycoproteins which mediate a variety of cell-cell and cell-substratum interactions. It is a constituent of the extracellular matrix of connective tissue and circulates in plasma. When suspension and adherent cultures of a human hepatoma cell line (SK-HEP-1) were incubated in serum-free medium, the resulting conditioned medium contained material which was specifically immunoprecipitated by antisera to human plasma fibronectin. By double immunodiffusion, a component in the conditioned culture medium was shown to form a line of identity with fibronectin in human plasma and to migrate as an alpha 2- to beta-globulin during immunoelectrophoresis. Human fibronectin was quantified in conditioned medium by electroimmunodiffusion, and was found to increase for at least three days at about 0.1 micrograms/10(6) cells/day. Adherent cultures of SK-HEP-1 cells were incubated with L-(/sup 35/S)methionine to label newly synthesized proteins. Labeled fibronectin in conditioned medium or in cell extracts comigrated with fibronectin in human plasma as shown by autoradiography following crossed-immunoelectrophoresis. Fibronectin was demonstrated in the extra-cellular matrix of adherent SK-HEP-1 cultures by immunofluorescence. It was shown previously that SK-HEP-1 cells synthesize alpha 1-protease inhibitor, one of the products of normal hepatocytes. The finding that these hepatoma cells also synthesize fibronectin supports the concept that the hepatocyte may be one source of circulating fibronectin, a possibility consistent with the established role of this cell type in blood plasma protein synthesis.

Glasgow, J.E.; Colman, R.W.



Establishment of a corneal epithelial cell line spontaneously derived from human limbal cells.  


The objective of this study was to establish a spontaneously derived human corneal epithelial cell line from a normal human limbus that retains differentiation potential and proliferative properties under continuous cell culture. After 50 passages of epithelial cells obtained from human limbal tissue a cell line spontaneously emerged. The immortalized cells showed a cobblestone appearance and displayed dense microvilli on their apical cell surface membrane. Colony forming efficiency was 5-6% and population doubling time was 19.6 h. In the mRNA level, cytokeratin (CK) 3 and 12 were detected in this cell line. In the protein level, the cells expressed CK3, CK12, CK14, CK19, vimentin, and some other proteins such as F-actin and beta-tubulin and beta(1)-integrin. They lacked p63. The immortalized cells had a heteroploid karyotype, but did not exhibit tumorigenic features. When cultured on an air-liquid interface the cells could form stratified multilayer epithelia. In summary, all these results indicated that a new human corneal epithelial cell line was spontaneously established from normal limbal tissue through serial culture. This cell line would be useful for studies of corneal epithelial biology and reconstructive corneal tissue engineering. PMID:17223104

Liu, Jingbo; Song, Ge; Wang, Zhichong; Huang, Bing; Gao, Qianying; Liu, Bingqian; Xu, Ying; Liang, Xuanwei; Ma, Ping; Gao, Nan; Ge, Jian



Aberrant autophosphorylation of c-Kit receptor in canine mast cell tumor cell lines  

Microsoft Academic Search

Several studies indicated that KIT mutation could cause ligand-independent activation of c-Kit receptor in canine mast cell tumor (MCT). The objective of this study was to investigate mechanisms of c-Kit receptor activation in various canine MCT cell lines. Four cell lines, HRMC (derived from cutaneous MCT), VIMC1 (visceral MCT), CoMS1 (visceral MCT) and CMMC1 (cutaneous MCT), were cultured in stem

Yoshinori Takeuchi; Yasuhito Fujino; Manabu Watanabe; Takayuki Nakagawa; Koichi Ohno; Nobuo Sasaki; Sumio Sugano; Hajime Tsujimoto



Characterization of human embryonic stem cell lines by the International Stem Cell Initiative  

Microsoft Academic Search

The International Stem Cell Initiative characterized 59 human embryonic stem cell lines from 17 laboratories worldwide. Despite diverse genotypes and different techniques used for derivation and maintenance, all lines exhibited similar expression patterns for several markers of human embryonic stem cells. They expressed the glycolipid antigens SSEA3 and SSEA4, the keratan sulfate antigens TRA-1-60, TRA-1-81, GCTM2 and GCT343, and the

Oluseun Adewumi; Behrouz Aflatoonian; Lars Ahrlund-Richter; Michal Amit; Gemma Beighton; Paul A Bello; Nissim Benvenisty; Lorraine S Berry; Simon Bevan; Barak Blum; Justin Brooking; Kevin G Chen; Andre B H Choo; Gary A Churchill; Marie Corbel; Ivan Damjanov; Jon S Draper; Petr Dvorak; Katarina Emanuelsson; Roland A Fleck; Angela Ford; Karin Gertow; Marina Gertsenstein; Paul J Gokhale; Rebecca S Hamilton; Ales Hampl; Lyn E Healy; Outi Hovatta; Johan Hyllner; Marta P Imreh; Joseph Itskovitz-Eldor; Jamie Jackson; Jacqueline L Johnson; Mark Jones; Kehkooi Kee; Benjamin L King; Barbara B Knowles; Majlinda Lako; Franck Lebrin; Barbara S Mallon; Daisy Manning; Yoav Mayshar; Ronald D G Mckay; Anna E Michalska; Milla Mikkola; Masha Mileikovsky; Stephen L Minger; Harry D Moore; Christine L Mummery; Andras Nagy; Norio Nakatsuji; Carmel M O'Brien; Steve K W Oh; Cia Olsson; Timo Otonkoski; Kye-Yoon Park; Robert Passier; Hema Patel; Minal Patel; Roger Pedersen; Martin F Pera; Marian S Piekarczyk; Renee A Reijo Pera; Benjamin E Reubinoff; Allan J Robins; Janet Rossant; Peter Rugg-Gunn; Thomas C Schulz; Henrik Semb; Eric S Sherrer; Henrike Siemen; Glyn N Stacey; Miodrag Stojkovic; Hirofumi Suemori; Jin Szatkiewicz; Tikva Turetsky; Timo Tuuri; Steineke van den Brink; Kristina Vintersten; Sanna Vuoristo; Dorien Ward; Thomas A Weaver; Lesley A Young; Weidong Zhang; Peter W Andrews



Gliotoxin-induced cytotoxicity in three salmonid cell lines: Cell death by apoptosis and necrosis  

Microsoft Academic Search

Epithelial (CHSE-214), fibroblast (RTG-2) and macrophage (RTS11) cell lines from Chinook salmon and rainbow trout were tested for their sensitivity to gliotoxin, a fungal metabolite. Gliotoxin treatment for 6 or 24 h caused cell viability to decrease in a dose-dependent manner, with effective concentrations (EC50s) being similar for the three cell lines but varying with exposure time. Under some exposure

S. J. DeWitte-Orr; N. C. Bols



Relation of cell proliferation to expression of peripheral benzodiazepine receptors in human breast cancer cell lines  

Microsoft Academic Search

Peripheral benzodiazepine receptor (PBR) agonist [3H]Ro5-4864 has been shown to bind with high affinity to the human breast cancer cell line BT-20. Therefore, we investigated different human breast cancer cell lines with regard to binding to [3H]Ro5-4864 and staining with the PBR-specific monoclonal antibody 8D7. Results were correlated with cell proliferation characteristics. In flow cytometric analysis, the estrogen receptor (ER)-negative

Anne Beinlich; Renate Strohmeier; Manfred Kaufmann; Herbert Kuhl



Characterization of Human Neuroblastoma Cell Lines Established before and after Therapy.  

National Technical Information Service (NTIS)

Six new cell lines have been established from human neuroblastomas. Cell line SMS-KAN, from primary tumor before therapy, and line SMS-KANR, from bone marrow after chemotherapy and radiotherapy, were established from the same patient. Cell lines SMS-KCN (...

C. P. Reynolds J. L. Beidler B. A. Spengler D. A. Reynolds R. A. Ross



Double Minute Chromosomes and the Homogeneously Staining Regions in Chromosomes of a Human Neuroblastoma Cell Line  

Microsoft Academic Search

Four human neuroblastoma cell lines were studied by chromosome banding techniques. All of the lines contained a marker chromosome with a long nonbanding homogeneously staining region (HSR). The HSR-containing chromosome differed in each line. One line contained two classes of cells: one with an HSR marker chromosome and the other with double minute chromosomes. Each cell had one of these

Gloria Balaban-Malenbaum; Fred Gilbert



Development of a pluripotent ES-like cell line from Asian sea bass (Lates calcarifer)--an oviparous stem cell line mimicking viviparous ES cells.  


We report a pluripotent embryonic stem cell-like cell line designated as SBES from blastula stage embryos of Asian sea bass (Lates calcarifer), which is an economically important cultivable and edible marine fish species in India. The SBES cells were cultured at 28 degrees C in Leibovitz L-15 medium supplemented with 20% fetal bovine serum without a feeder layer. The ES-like cells were round or polygonal and grew exponentially in culture. The SBES cells exhibited an intense alkaline phosphatase activity and expression of transcription factor Oct 4. The undifferentiated state of these cells was maintained at low seeding densities and the cells formed embryoid bodies when seeded in bacteriological plates. On treatment with all-trans retinoic acid, these cells differentiated into neuron-like cells, muscle cells, and beating cardiomyocytes, indicating their pluripotency. This embryonic ES-like cell line derived from an oviparous fish blastula conserved several peculiar features of viviparous mammalian embryonic stem cell lines. The present study highlights the importance and potential of piscine ES-like cell line for stem cell research without evoking ethical issues and invasive interventions sparing mammalian embryos. PMID:17704967

Parameswaran, V; Shukla, Ravi; Bhonde, Ramesh; Hameed, A S Sahul



Characteristics of bovine inner cell mass-derived cell lines and their fate in chimeric conceptuses.  


Bovine embryonic stem (ES) cells have the potential to provide significant benefits in a range of agricultural and biomedical applications. Here, we employed a combination of conventional methods using glycogen synthase kinase 3 and mitogen-activated protein kinase inhibitors to establish ES cell lines from in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) bovine embryos. Five male cell lines were established from IVF embryos, and two female and three male cell lines from SCNT blastocysts; we named these lines bovine ES cell-like cells (bESLCs). The lines exhibited dome-shaped colonies, stained positively for alkaline phosphatase, and expressed pluripotent stem cell markers such as POU5F1, SOX2, and SSEA-1. The expression levels of these markers, especially for NANOG, varied among the cell lines. A DNA methylation assay showed the POU5F1 promoter region was hypomethylated compared to fibroblast cells. An in vitro differentiation assay showed that endoderm and ectoderm marker genes, but not mesoderm markers, were upregulated in differentiating bESLCs. To examine bESLCs in later embryonic stages, we created 22 chimeric blastocysts with a male bESLC line carrying a GFP marker gene and transferred these to a recipient cow. Four chimeric embryos were subsequently retrieved on Day 13 and retransferred to two recipient cows. One living fetus was obtained at Day 62. GFP signals were not identified in fetal cells by fluorescence microscopy; however, genomic PCR analysis detected the GFP gene in major organs. Clusters of GFP-positive cells were observed in amniotic membranes, suggesting that bESLCs can be categorized as a novel type of ICM-derived cells that can potentially differentiate into epiblast and hypoblast lineages. PMID:23782837

Furusawa, Tadashi; Ohkoshi, Katsuhiro; Kimura, Koji; Matsuyama, Shuichi; Akagi, Satoshi; Kaneda, Masahiro; Ikeda, Mitsumi; Hosoe, Misa; Kizaki, Keiichiro; Tokunaga, Tomoyuki



Can we develop ethically universal embryonic stem-cell lines?  

Microsoft Academic Search

Human embryonic stem-cell (hESC) research faces opposition from those who object to the destruction of human embryos. Over the past few years, a series of new approaches have been proposed for deriving hESC lines without injuring a living embryo. Each of these presents scientific challenges and raises ethical and political questions. Do any of these methods have the potential to

Ronald M Green



Gypsy moth cell lines divergent in viral susceptibility  

Microsoft Academic Search

Summary  A series of cell lines unique in insect virus susceptibility pattern have been isolated from the ovaries of the gypsy moth\\u000a (Lymantria dispar: Lepidoptera: Lymantriidae) on a synthetic medium with mammalian and avian serum supplementation. Growth curves showed the\\u000a poorest growth occurring on peptone-based media with somewhat better growth on amino-acid-based media. The best growth was\\u000a obtained with combined media.

R. H. Goodwin; G. J. Tompkins; P. McCawley



KIR Receptor-Ligand Incompatibility Predicts Killing of Osteosarcoma Cell Lines by Allogeneic NK Cells  

PubMed Central

Background The effectiveness of KIR incompatible, alloreactive NK cells has been primarily documented in hematological malignancies following stem cell transplant. This effect has not been thoroughly evaluated for pediatric solid tumors. In this study, we evaluated KIR receptor-ligand incompatibility of NK cells against osteosarcoma cell lines. Procedure Following the KIR receptor-ligand mismatch model, MHC I cell surface expression and KIR ligand mRNA content of 3 osteosarcoma cell lines was determined by flow cytometry and quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR), respectively. NK cells were isolated from healthy volunteer donor peripheral blood mononuclear cells (PBMCs) and KIR surface expression determined by flow cytometry. An Annexin-V based flow cytometric killing assay was used to determine % of dying osteosarcoma target cells by donor NK effector cells. Results One of 7 healthy volunteer donors tested lacked phenotypic expression of one KIR. However, variable expression of KIR ligands was observed in 3 osteosarcoma cell lines. The highest rates of dying cells were seen in osteosarcoma cells with the lowest KIR ligand expression. Following down-regulation of KIR ligand expression, an increased susceptibility to NK cell mediated killing was observed in a previously NK-resistant osteosarcoma cell line. Conclusions Variable MHC I and KIR ligand expression was observed in osteosarcoma cell lines and this resulted in variable susceptibility to NK cell mediated killing predicted by the degree of KIR receptor-ligand incompatibility. Collectively, these data provide rationale for the study of KIR incompatible stem cell transplant for osteosarcoma, although further studies with fresh osteosarcoma samples are necessary.

Delgado, David; Webster, Daniel E.; DeSantes, Kenneth B.; Durkin, Emily T.; Shaaban, Aimen F.



Toxicological evaluation of magnetic ionic liquids in human cell lines.  


Magnetic ionic liquids (MILs) are new solvents with an interesting broad of applications however their toxicity is still an open issue. In this paper we report the toxicity of [C(8)MIM] and [Choline-C(n)] based magnetic ionic liquids assessed in two human cell lines: normal skin fibroblasts (CRL-1502) and colorectal adenocarcinoma cells (CaCo-2), acquiring this last characteristics of human enterocytes after differentiation. The results showed that [CoCl(4)] and [MnCl(4)] are more prone to generate cytotoxicity. PMID:23561571

Frade, Raquel F M; Simeonov, Svilen; Rosatella, Andreia A; Siopa, Filipa; Afonso, Carlos A M



Identification of a FXIIIA variant in human neuroblastoma cell lines  

PubMed Central

FXIII is a transglutaminase consisting of two catalytic (FXIIIA) and two non-catalytic subunits (FXIIIB) in plasma, where this enzyme is responsible for stabilizing fibrin clots. Although possible functions of intracellular FXIIIA have been proposed, these remain to be established. We show that a 40 kDa protein species of FXIIIA is present in the human neuroblastoma cell lines SH-SY5Y and LAN5. These data reveal the presence of a new uncharacterised variant of FXIIIA, possibly due to an alternative splicing, in nervous cells.

lannaccone, Martina; Giuberti, Gaia; Vivo, Giulia De; Caraglia, Michele; Gentile, Vittorio



Sigma 1 binding in a human neuroblastoma cell line  

Microsoft Academic Search

Behaviorally, sigma1 agents modulate opioid analgesia. To examine possible mechanisms responsible for these interactions, we have identified a\\u000a cell line containing both sigma1 and opioid receptors. [3H](+)-pentazocine binding in BE(2)-C human neuroblastoma cells is high affinity (KD 3.4±0.7 nM) and high density (Bmax 2.98±0.14 pmol\\/mg protein). Competition studies reveal a selectivity profile similar to that of sigma1 sites in guinea

Jennifer Ryan-Moro; Chih-Cheng Chien; Kelly M. Standifer; Gavril W. Pasternak



icb-1 Gene counteracts growth of ovarian cancer cell lines.  


Human gene icb-1 has been originally identified to be involved in differentiation processes of cancer cells. To examine the function of icb-1 in ovarian cancer, we knocked down its expression in three ovarian cancer cell lines and performed microarray-based gene expression profiling with subsequent gene network modeling. Loss of icb-1 expression accelerated proliferation of SK-OV-3, OVCAR-3 and OAW-42 cells and led to upregulation of ovarian cancer biomarkers like KLK10 and CLDN16. Most of the upregulated genes were part of oncogenic pathways regulated by ER? or TNF. Our data suggest that icb-1 gene inhibits growth and progression of ovarian cancer cells. PMID:23474491

Treeck, Oliver; Schüler, Susanne; Häring, Julia; Skrzypczak, Maciej; Lattrich, Claus; Ortmann, Olaf



New cell lines from mouse epiblast share defining features with human embryonic stem cells.  


The application of human embryonic stem (ES) cells in medicine and biology has an inherent reliance on understanding the starting cell population. Human ES cells differ from mouse ES cells and the specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo before implantation in the uterus. Here we show that cell lines can be derived from the epiblast, a tissue of the post-implantation embryo that generates the embryo proper. These cells, which we refer to as EpiSCs (post-implantation epiblast-derived stem cells), express transcription factors known to regulate pluripotency, maintain their genomic integrity, and robustly differentiate into the major somatic cell types as well as primordial germ cells. The EpiSC lines are distinct from mouse ES cells in their epigenetic state and the signals controlling their differentiation. Furthermore, EpiSC and human ES cells share patterns of gene expression and signalling responses that normally function in the epiblast. These results show that epiblast cells can be maintained as stable cell lines and interrogated to understand how pluripotent cells generate distinct fates during early development. PMID:17597760

Tesar, Paul J; Chenoweth, Josh G; Brook, Frances A; Davies, Timothy J; Evans, Edward P; Mack, David L; Gardner, Richard L; McKay, Ronald D G



Cell-type-dependent thyroid hormone effects on glioma tumor cell lines.  


Purpose. The present study investigated the potential effects of long-term T3 treatment on glioma tumor cell lines. Thyroid hormone action on cell growth, differentiation and survival during development may be of therapeutic relevance Methods and Results 1321N1 cell line, an astrocytoma grade II, and U87MG, a glioblastoma grade IV, were exposed for 2 and 4 days in medium deprived of T3 and in medium containing 1?nM T3. T3 promoted re-differentiation in both cell lines. However, T3 increased cell proliferation in 1321N1 (2?days) which declined thereafter (4?days) while in U87MG resulted in suppression of cell proliferation. At the molecular level, a 2.9 fold increase in the expression of TR?1 receptor was observed in U87MG versus 1321N1, P < 0.05. TR?1 receptor was undetectable. These changes corresponded to a distinct pattern of T3-induced kinase signaling activation; T3 had no effect on ERK activation in both cell lines but significantly increased phospho-Akt levels in 1321N1. Conclusion. In conclusion, T3 can re-differentiate glioma tumor cells, whereas its effect on cell proliferation appears to be dependent on the type of tumor cell line with aggressive tumors being more sensitive to T3. TR?1 receptor may, at least in part, be implicated in this response. PMID:22229106

Liappas, Alexandros; Alexandros, Liappas; Mourouzis, Iordanis; Iordanis, Mourouzis; Zisakis, Athanasios; Athanasios, Zisakis; Economou, Konstantinos; Konstantinos, Economou; Lea, Robert-William; Robert-William, Lea; Pantos, Constantinos; Constantinos, Pantos



Cell-Type-Dependent Thyroid Hormone Effects on Glioma Tumor Cell Lines  

PubMed Central

Purpose. The present study investigated the potential effects of long-term T3 treatment on glioma tumor cell lines. Thyroid hormone action on cell growth, differentiation and survival during development may be of therapeutic relevance Methods and Results 1321N1 cell line, an astrocytoma grade II, and U87MG, a glioblastoma grade IV, were exposed for 2 and 4 days in medium deprived of T3 and in medium containing 1?nM T3. T3 promoted re-differentiation in both cell lines. However, T3 increased cell proliferation in 1321N1 (2?days) which declined thereafter (4?days) while in U87MG resulted in suppression of cell proliferation. At the molecular level, a 2.9 fold increase in the expression of TR?1 receptor was observed in U87MG versus 1321N1, P < 0.05. TR?1 receptor was undetectable. These changes corresponded to a distinct pattern of T3-induced kinase signaling activation; T3 had no effect on ERK activation in both cell lines but significantly increased phospho-Akt levels in 1321N1. Conclusion. In conclusion, T3 can re-differentiate glioma tumor cells, whereas its effect on cell proliferation appears to be dependent on the type of tumor cell line with aggressive tumors being more sensitive to T3. TR?1 receptor may, at least in part, be implicated in this response.

Alexandros, Liappas; Iordanis, Mourouzis; Athanasios, Zisakis; Konstantinos, Economou; Robert-William, Lea; Constantinos, Pantos



Analysis of Cell Surface N-glycosylation of the Human Embryonic Kidney 293T Cell Line  

Microsoft Academic Search

Protein glycosylation is a prominent posttranslational modification and is involved in many biological functions. Human cell lines used for the expression of recombinant glycoproteins present variations in their cell surface N-glycosylation due to their cell type–specific origin. We therefore investigated the presence of specific glycosyltransferases by RT-PCR and the cell surface N-glycan structures of HEK293T cells by MALDI-TOF-MS and MALDI-TOF\\/TOF-MS

Stefan O. Reinke; Marion Bayer; Markus Berger; Véronique Blanchard; Stephan Hinderlich



Coumarin modulates the cell-cycle progression of an MTV-EJras cell line  

Microsoft Academic Search

The effects of coumarin (1,2-benzopyrone) onras oncogene expression during the cell cycle of an MTV-EJras cell line was determined by flow cytometry.ras oncogene expression in cells was induced by dexamethasone and increased fivefold during G1\\/G0 phase and threefold in S phase. Dexamethasone also increased the percentage of cells in S phase from 21% to 31%, compared to phosphate-buffered-saline-treated control cells

J. Kahn; P. Preis; F. Waldman; A. Tseng Jr



Cytotoxic Activity of New Acetoxycoumarin Derivatives in Cancer Cell Lines  

PubMed Central

Background Coumarin and their derivatives are important and useful compounds with diverse pharmacological properties. In the present study, we evaluated the in vitro cytotoxic activity of new acetoxycoumarin derivatives: 4-(7-methoxy-4-methyl-2-oxo-2H-chromen-3-yl)phenyl acetate (1), 4-(1-methyl-3-oxo-3H-benzo[f]chromen-2-yl)phenyl acetate (2), 4-(6-propionamido-4-methyl-2-oxo-2H-chromen-3-yl)phenyl acetate (3), 4-(7-acetoxy-2-oxo-4-phenyl-2H-chromen-3-yl)phenyl acetate (4), 4-(2-oxo-4-phenyl-2H-chromen-3-yl)phenyl acetate (5), 4-(6-bromo-2-oxo-4-phenyl-2H-chromen-3-yl)phenyl acetate (6), 4-(7-(diethylamino)-4-methyl-2-oxo-2H-chromen-3-yl)phenyl acetate (7), 4-(6,8-dibromo-4-methyl-2-oxo-2H-chromen-3-yl)phenyl acetate (8) against A549 human lung cancer, CRL 1548 rat liver cancer and CRL 1439 normal rat liver cells. Materials and Methods The cytotoxic activity was evaluated by crystal violet dye-binding assay. The effect of compounds 5 and 7 on different phases of the cell cycle was determined using flow cytometry. Results In the A549 lung cancer cell line, the 50% lethal dose (LD50) values for compounds 1–4, 6 and 8 were found to be >100 ?M while those for 5 and 7 were 89.3 and 48.1 ?M, respectively after 48 h treatment. In the CRL 1548 liver cancer cell line, only compound 7 showed toxicity, with an LD50 of 45.1 ?M. Compounds 5 and 7 caused different cell phase arrest in lung and liver cancer cell lines. Conclusion The results indicate that 4-(7-(diethylamino)-4-methyl-2-oxo-2H-chromen-3-yl)phenyl acetate (7) had the highest cytotoxic activity in all of the examined cell lines.

Musa, Musiliyu A.; Badisa, Veera L. D.; Latinwo, Lekan M.; Cooperwood, John; Sinclair, Andre; Abdullah, Ahkinyala



Primed pluripotent cell lines derived from various embryonic origins and somatic cells in pig.  


Since pluripotent embryonic stem cell (ESC) lines were first derived from the mouse, tremendous efforts have been made to establish ESC lines in several domestic species including the pig; however, authentic porcine ESCs have not yet been established. It has proven difficult to maintain an ESC-like state in pluripotent porcine cell lines due to the frequent occurrence of spontaneous differentiation into an epiblast stem cell (EpiSC)-like state during culture. We have been able to derive EpiSC-like porcine ESC (pESC) lines from blastocyst stage porcine embryos of various origins, including in vitro fertilized (IVF), in vivo derived, IVF aggregated, and parthenogenetic embryos. In addition, we have generated induced pluripotent stem cells (piPSCs) via plasmid transfection of reprogramming factors (Oct4, Sox2, Klf4, and c-Myc) into porcine fibroblast cells. In this study, we analyzed characteristics such as marker expression, pluripotency and the X chromosome inactivation status in female of our EpiSC-like pESC lines along with our piPSC line. Our results show that these cell lines demonstrate the expression of genes associated with the Activin/Nodal and FGF2 pathways along with the expression of pluripotent markers Oct4, Sox2, Nanog, SSEA4, TRA 1-60 and TRA 1-81. Furthermore all of these cell lines showed in vitro differentiation potential, the X chromosome inactivation in female and a normal karyotype. Here we suggest that the porcine species undergoes reprogramming into a primed state during the establishment of pluripotent stem cell lines. PMID:23326334

Park, Jin-Kyu; Kim, Hye-Sun; Uh, Kyung-Jun; Choi, Kwang-Hwan; Kim, Hyeong-Min; Lee, Taeheon; Yang, Byung-Chul; Kim, Hyun-Jong; Ka, Hak-Hyun; Kim, Heebal; Lee, Chang-Kyu



Investigation of native fluorescence spectral difference among prostate cancer cell lines with different risk levels  

NASA Astrophysics Data System (ADS)

The alteration of native fluorophores among different types of cancer cell lines was investigated by the fluorescence spectroscopy. Different types of cancer cell lines with different risk levels, such as moderate metastatic (DU-145) and advanced metastatic (PC-3) cell lines as well as normal cell line (Fibroblast), were excited by the selective excitation wavelength of 300 nm to explore changes of the relative contents of tryptophan and NADH using principal component analysis (PCA). The higher relative content of tryptophan was observed in the advanced metastatic cancer cell lines in comparison with the moderate metastatic and non aggressive cell lines.

Pu, Yang; Xue, Jianpeng; Xu, Baogang; Wang, Wubao; Gu, Yueqing; Tang, Rui; Achilefu, S.; Ackerstaff, Ellen; Koutcher, Jason A.; Alfano, R. R.


Differentiation Properties of Avian Tumor Cell Lines: Analysis Using Chicken Fetal Antigen and Other Markers1  

Microsoft Academic Search

Three avian lymphoblastoid tumor cell lines were compared for the expression of the serologically complex cell surface antigen, chicken fetal antigen (CFA), and other properties asso ciated with differentiation. Two Marek's disease cell lines (CU- 12 and CU-36) and a cell line associated with lymphoid leukosis (CU-10) were all found to differ qualitatively in the expression of CFA. While both

Kathryn A. Trembicki; Muquarrab A. Qureshi; Rodney R. Dieted


Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line  

Microsoft Academic Search

In contrast to mouse epidermal cells, hu- man skin keratinocytes are rather resistant to transfor- mation in vitro. Immortalization has been achieved by SV40 but has resulted in cell lines with altered differentiation. We have established a spontaneously transformed human epithelial cell line from adult skin, which maintains full epidermal differentiation capacity. This HaCaT cell line is obviously immortal (>140

Petra Boukamp; T. Petrussevska; Dirk Breitkreutz; Jiirgen Hornung; Alex Markham; Norbert E. Fusenig



Hormone Sensitivity is Reflected in the Phospholipid Profiles of Breast Cancer Cell Lines  

Microsoft Academic Search

We have found that the profiles of total phospholipids in malignant breast cancer cell lines change going from hormone sensitive to highly hormone resistant cells lines. In particular, two phospholipid components that were absent or at very low levels in hormone sensitive MCF7 cells and moderately hormone sensitive cell lines (MIII, LCC2) were found in relatively high proportions in highly

Marina Sterin; Jack S. Cohen; Israel Ringel



Effects of Lipopolysaccharide on Newly Established Rat Dental Pulp–derived Cell Line with Odontoblastic Properties  

Microsoft Academic Search

To clarify mechanisms of pulp wound healing and regeneration, it is important to establish continuous odontoblast-lineage cell lines. In this study, we established the proliferating pulp progenitor cell lines from dental papilla cells of rat incisor. These cell lines showed high levels of alkaline phosphatase (ALP) activity, expression of Runx2 and dentin sialophosphoprotein (DSPP), and extracellular formation of mineralized nodules.

Kimiko Nomiyama; Chiaki Kitamura; Toshiyuki Tsujisawa; Masato Nagayoshi; Takahiko Morotomi; Masmichi Terashita; Tatsuji Nishihara



Antiproliferative effect of Tualang honey on oral squamous cell carcinoma and osteosarcoma cell lines  

Microsoft Academic Search

BACKGROUND: The treatment of oral squamous cell carcinomas (OSCC) and human osteosarcoma (HOS) includes surgery and\\/or radiotherapy which often lead to reduced quality of life. This study was aimed to study the antiproliferative activity of local honey (Tualang) on OSCC and HOS cell lines. METHODS: Several concentrations of Tualang honey (1% - 20%) were applied on OSCC and HOS cell

Abdulmlik A Ghashm; Nor H Othman; Mohammed N Khattak; Noorliza M Ismail; Rajan Saini



The Establishment of a Cell Line of Human Hormone-synthesizing Trophoblastic Cells in Vitro1  

Microsoft Academic Search

SUMMARY A human hormone-synthesizing trophoblastic cell system has been established in vitro and may prove to be the first func tional human embryonic cell line in continuous culture. Cho- rionic gonadotropin hormone produced by these cultures serve as a marker for identification of the trophoblastic cell. No interruption in this property nor change in cytologie display has occurred during 1.5

Roland A. Pattillo; George O. Gey



Suppression of Tumorigenicity in Human Cell Hybrids Derived from Cell Lines Expressing Different Activated ras Oncogenes  

Microsoft Academic Search

Four different human tissue-derived cell lines, each previously shown to express either a Ha-, Ki-, or N-ras-activated oncogene, were fused in four different paired combinations. The three combinations that involved the tumor line HT1080 (activated N-ras oncogene) were found to be tumorigenic in nude mice, but to different degrees. However, the fusion of the tumor lines EJ and SW480 (activated

Andrew G. Geiser; Michael J. Anderson; Eric J. Stanbridge



Cell Type-specific ?2-Adrenergic Receptor Clusters Identified Using Photoactivated Localization Microscopy Are Not Lipid Raft Related, but Depend on Actin Cytoskeleton Integrity*  

PubMed Central

Recent developments in the field of optical super-resolution techniques allow both a 10-fold increase in resolution as well as an increased ability to quantify the number of labeled molecules visualized in the fluorescence measurement. By using photoactivated localization microscopy (PALM) and an experimental approach based on the systematic comparison with a nonclustering peptide as a negative control, we found that the prototypical G protein-coupled receptor ?2-adrenergic receptor is partially preassociated in nanoscale-sized clusters only in the cardiomyocytes, such as H9C2 cells, but not in other cell lines, such as HeLa and Chinese hamster ovary (CHO). The addition of the agonist for very short times or the addition of the inverse agonist did not significantly affect the organization of receptor assembly. To investigate the mechanism governing cluster formation, we altered plasma membrane properties with cholesterol removal and actin microfilament disruption. Although cholesterol is an essential component of cell membranes and it is supposed to be enriched in the lipid rafts, its sequestration and removal did not affect receptor clustering, whereas the inhibition of actin polymerization did decrease the number of clusters. Our findings are therefore consistent with a model in which ?2 receptor clustering is influenced by the actin cytoskeleton, but it does not rely on lipid raft integrity, thus ruling out the possibility that cell type-specific ?2 receptor clustering is associated with the raft.

Scarselli, Marco; Annibale, Paolo; Radenovic, Aleksandra



Cell type-specific ?2-adrenergic receptor clusters identified using photoactivated localization microscopy are not lipid raft related, but depend on actin cytoskeleton integrity.  


Recent developments in the field of optical super-resolution techniques allow both a 10-fold increase in resolution as well as an increased ability to quantify the number of labeled molecules visualized in the fluorescence measurement. By using photoactivated localization microscopy (PALM) and an experimental approach based on the systematic comparison with a nonclustering peptide as a negative control, we found that the prototypical G protein-coupled receptor ?2-adrenergic receptor is partially preassociated in nanoscale-sized clusters only in the cardiomyocytes, such as H9C2 cells, but not in other cell lines, such as HeLa and Chinese hamster ovary (CHO). The addition of the agonist for very short times or the addition of the inverse agonist did not significantly affect the organization of receptor assembly. To investigate the mechanism governing cluster formation, we altered plasma membrane properties with cholesterol removal and actin microfilament disruption. Although cholesterol is an essential component of cell membranes and it is supposed to be enriched in the lipid rafts, its sequestration and removal did not affect receptor clustering, whereas the inhibition of actin polymerization did decrease the number of clusters. Our findings are therefore consistent with a model in which ?2 receptor clustering is influenced by the actin cytoskeleton, but it does not rely on lipid raft integrity, thus ruling out the possibility that cell type-specific ?2 receptor clustering is associated with the raft. PMID:22442147

Scarselli, Marco; Annibale, Paolo; Radenovic, Aleksandra



Aberrant autophosphorylation of c-Kit receptor in canine mast cell tumor cell lines.  


Several studies indicated that KIT mutation could cause ligand-independent activation of c-Kit receptor in canine mast cell tumor (MCT). The objective of this study was to investigate mechanisms of c-Kit receptor activation in various canine MCT cell lines. Four cell lines, HRMC (derived from cutaneous MCT), VIMC1 (visceral MCT), CoMS1 (visceral MCT) and CMMC1 (cutaneous MCT), were cultured in stem cell factor (SCF, a ligand of c-Kit receptor)-free medium and subjected to analyses of KIT mutation, c-Kit receptor phosphorylation, SCF expression and the effects of SCF stimulation. In addition, the SCF/c-Kit receptor autocrine mechanism was verified in HRMC cells. HRMC cells expressed wild type c-Kit receptor. Both VIMC1 and CoMS1 cells had the same one amino acid (AA) substitution in the extracellular domain of c-Kit receptor. CMMC1 cells had at least three variants of c-Kit receptor such as one AA deletion in the extracellular domain (variant A), one AA substitution in the extracellular domain as well as an internal tandem duplication in the juxtamembrane domain (variant B), and a nonsense mutation (variant C). Both mature and immature forms of c-Kit receptor were observed and the c-Kit receptors were phosphorylated in all cell lines. While both mature and immature forms of c-Kit receptor were substantially phosphorylated in CMMC1 cells, the immature form was slightly phosphorylated in other cell lines. Phosphorylation of c-Kit receptor in HRMC, VIMC1 and CoMS1 cells were enhanced by SCF stimulation whereas no enhancement was observed in CMMC1 cells. There was no effect of SCF stimulation on proliferation of all the cell lines. SCF protein was detectable in only HRMC cells although mRNA expression of SCF was detected in all the cell lines. A tyrosine kinase inhibitor Dasatinib (internal inhibitor) inhibited c-Kit receptor phosphorylation in HRMC cells whereas anti-canine SCF antibody (external inhibitor) had no inhibitory effect. Thus there could be no external SCF/c-Kit receptor autocrine mechanism whereas there could be an internal autocrine mechanism within HRMC cells. The results indicated that consistent c-Kit receptor phosphorylation could be caused by the stimulation with autocrine SCF in HRMC cells while it could be caused by functional mutations of KIT in VIMC1, CoMS1 and CMMC1 cells. As the four canine MCT cell lines had various aberrations associated with c-Kit receptor phosphorylation, KIT mutation and SCF expression, such molecular biological diversity might reflect the different biological behavior in canine MCT. PMID:20591500

Takeuchi, Yoshinori; Fujino, Yasuhito; Watanabe, Manabu; Nakagawa, Takayuki; Ohno, Koichi; Sasaki, Nobuo; Sugano, Sumio; Tsujimoto, Hajime



Determinants of the sensitivity of human small-cell lung cancer cell lines to methotrexate.  

PubMed Central

We have characterized the determinants of methotrexate (MTX) responsiveness in eight patient-derived cell lines of small-cell lung cancer (SCLC). Clonogenic survival was correlated with factors known to affect sensitivity to drug. NCI-H209 and NCI-H128 were most drug sensitive, with drug concentrations required to inhibit clonogenic survival by 50% with less than 0.1 microM MTX. Six cell lines (NCI-H187, NCI-H345, NCI-H60, NCI-H524, NCI-H146, and NCI-N417D) were relatively drug resistant. In all cell lines studied, higher molecular weight MTX-polyglutamates (MTX-PGs) with 3-5 glutamyl moieties (MTX-Glu3 through MTX-Glu5) were selectively retained. Relative resistance to low (1.0 microM) drug concentrations appeared to be largely due to decreased intracellular metabolism of MTX. Five of the six resistant lines were able to synthesize polyglutamates at higher (10 microM) drug concentrations, although one resistant cell line (NCI-N417D) did not synthesize higher molecular weight MTX-PGs, even after exposure to 10 microM drug. Two cell lines with resistance to 10 microM MTX (NCI-H146 and NCI-H524) synthesized and retained higher molecular weight MTX-PGs in excess of binding capacity after exposure to 10 microM drug. However, the specific activity of thymidylate synthase in these cell lines was low. MTX sensitivity in patient-derived cell lines of SCLC requires the ability of cells to accumulate and retain intracellular drug in the form of polyglutamate metabolites in excess of dihydrofolate reductase, as well as a high basal level of consumption of reduced folates in the synthesis of thymidylate.

Curt, G A; Jolivet, J; Carney, D N; Bailey, B D; Drake, J C; Clendeninn, N J; Chabner, B A



Multiple activities of a cloned cell line mediating natural killer cell function.  


A special class of immunologic cells can lyse or damage a variety of target cells, notably malignant cells in vitro. These cells have been called natural killer (NK) cells because lysis does not require deliberate immunization by tumor cells. Although these cells can be distinguished from conventional T cells, B cells, and phagocytic cells, they have been difficult to define. We describe a representative cloned cell line that was obtained by cloning Ig -Ly-5+ cells from spleen. This clone, Cl.Ly-1-2-NK-1+/11, displays Thy-1, Ly-5, Qat-4, Qat-5 and NK-1 cell surface antigens and lyses the NK-sensitive YAC-1 lymphoma cells, but does not lyse RL-12 cells, an NK-resistant lymphoma. In addition, this clone lysed the P815 mastocytoma, EL4 lymphoma, and lipopolysaccharide-activated B lymphocyte targets. This cloned population therefore combined information for a unique display of cell surface antigens and specialized function similar to "activated" NK cells. Because this cloned population forms conjugates with susceptible but not resistant target cells, it may prove useful to identify the structure of cell surface molecules that recognize foreign cells. Finally, cells of this clone also specificity lysed target cells coated by antibodies to determinants on the target cell surface, demonstrating that a single cloned cell population can mediate two specialized immunologic functions: antibody-dependent cellular cytotoxicity and NK cell lysis. PMID:6973001

Nabel, G; Bucalo, L R; Allard, J; Wigzell, H; Cantor, H



Fluorouracil Selectively Enriches Stem-like Leukemic Cells in a Leukemic Cell Line  

PubMed Central

Recent studies have reported that cancer stem cells (CSCs) could be isolated from solid cancer cell lines, in which the purity of CSCs was higher than that from tumor tissues. Separation of CSCs from leukemic cell lines was rarely reported. In this study, CD34+CD38- stem-like cell subsets in human KG-1a leukemic cell line were enriched by cytotoxic agent 5-fluorouracil (5-FU). After 4 days incubation of KG-1a cell line with 5-FU (50 ?g/ml), the CD34+CD38- subpopulation of cell lines was enriched more than 10 times. The enriched cells had proliferate potential in vitro, low level of RNA transcription and Hoechst 33342 dye efflux ability, accompanied by high expression of ATP-binding cassette transporter protein ABCG2. Our findings suggest that treatment with 5-FU offers an easy method to isolate leukemic stem-like subpopulation. It can facilitate studies of leukemic stem cell biology and the development of new therapeutic strategies.

Zhang, Ling; Yang, Song; He, Yu-Juan; Shao, Hui-Yuan; Wang, Li; Chen, Hui; Gao, Yu-Jie; Qing, Feng-Xian; Chen, Xian-Chun; Zhao, Liu-Yang; Tan, Shi



Mechanisms of prodigiosin cytotoxicity in human neuroblastoma cell lines.  


Prodigiosin is a bacterial red pigment with cytotoxic properties and potential antitumor activity that has been tested against different cancerous cells. In this study we report the effect and mechanisms of action of prodigiosin against different human neuroblastoma cell lines: SH-SY5Y, LAN-1, IMR-32 (N-type) and SK-N-AS (S-type). We compare the anticancerous effect of prodigiosin with that of cisplatin at different concentrations during 24 h of exposure. Prodigiosin is more potent, with IC50 values lower than 1.5 microM in N-type neuroblastoma cells and around 7 microM in the S-type neuroblastoma cell line. We describe prodigiosin as a proton sequestering agent that destroys the intracellular pH gradient, and propose that its main cytotoxic effect could be related to its action on mitochondria, where it exerts an uncoupling effect on the electronic chain transport of protons to mitochondrial ATP synthase. As a result of this action, ATP production is reduced but without decreasing in oxygen consumption. This mechanism of action differs from those induced by conventional chemotherapeutic drugs, suggesting a possible role for prodigiosin to enhance the effect of antitumor agents in the treatment of neuroblastoma. PMID:17678643

Francisco, Roser; Pérez-Tomás, Ricardo; Gimènez-Bonafé, Pepita; Soto-Cerrato, Vanessa; Giménez-Xavier, Pol; Ambrosio, Santiago



Altered Topoisomerase Ila in a Drug-resistant Small Cell Lung Cancer Cell Line Selected in VP-161  

Microsoft Academic Search

The H209\\/V6 cell line was derived from the H209 small cell lung cancer cell line by selection in etoposide (VP-16). Cytogenetic analysis indicates that the sensitive and resistant cell lines share 20 marker chromosomes and thus are clearly related. However, the H209\\/V6 cell line has four additional structurally altered chromosomes and a 2 N-modal chromo some number, while the H209

Shelagh E. L. Mirski; Cindy D. Evans; Kurt C. Almquist; Marilyn L. Slovak; Susan P. C. Cole


Characterization and chemosensitivity of two cell lines derived from human glioblastomas  

Microsoft Academic Search

Summary We have characterized two human glioblastoma cell lines, which were designated as YH cells and AM cells. The two cell lines maintained morphological appearance observed in the primary culture and immunohis-tochemically expressed glial fibrillary acidic protein (GFAP) and S-100 protein. Population doubling time for YH cells and AM cells indicated 30 hours and 25 hours, respectively, in an exponential

Ichiro Izumu; Katsuyoshi Mineura; Katsuo Watanabe; Masayoshi Kowada



Influence of 864 MHz electromagnetic field on growth kinetics of established cell line  

Microsoft Academic Search

Considering often contradictory data on biological effects of mobile phones frequencies on established cell culture lines, our study aimed at evaluating the influence of 864 MHz electromagnetic field on proliferation, colony forming ability and viability of Chinese hamster lung cells of line V79 cell. Prior to exposure for 1, 2 and 3 hours in transversal electromagnetic mode cell (TEM-cell) cell

Ivan Pavicic; Ivancica Trosic



Differences in radiation response among human cervix carcinoma cell lines.  


The radiobiological properties of five newly established carcinoma of the cervix cell lines have been determined. By means of an in vitro clonogenic assay, survival curves were compared at radiation dose rates of 150, 3.2 and 1.6 cGy/min. In terms of both acute radiosensitivity and the extent of low dose-rate sparing, large differences existed among the lines. Two of the lines, HX151c and HX160c, were radiosensitive (surviving fractions at 2 Gy of 0.23 and 0.33 respectively) and showed little radiation recovery capacity during protracted irradiation, whereas the remaining three lines had much higher surviving fraction at 2 Gy (up to 0.6) and showed considerable low dose-rate sparing. The data were well fitted by the Incomplete Repair model of Thames; repair half times ranged from 0.25 to 5.7 h. These findings indicate that large differences in intrinsic radiation recovery capacity exist within this tumour type, differences which emphasise that predictive testing of intrinsic radiosensitivity may be of clinical value in this disease. PMID:3222467

Kelland, L R; Steel, G G



Isolation and characterization of cancer stem like cells in human glioblastoma cell lines.  


To identify and compare the features of stem like cells in human glioblastoma cell lines U251, U87MG, A172 with primary cultured glioblastoma stem cells, the ratio of CD133+ cells, the ability of tumor sphere formation, and self-renewing capacity of U251, U87MG, A172 cells in serum free medium plus EGF, bFGF and B27 supplement were detected. The results suggested that there might be more cancer stem like cells in U251 cells compared with others. CD133+ cells enriched in SP cells and in U251 cells cultured with the serum free medium. They expressed the neural stem cell markers CD133 and Nestin, but lacked of neuronal and astrocyte marker MAP2, beta-III tubulin and GFAP. They could apparently generate both neurons and glial cells after serum retrieved in vitro. Gli1, Bmi1, Notch2 and PTEN were also found expressed highly in them. Moreover, CD133+ cells were more resistant to hypoxia, irradiations and some chemotherapeutics than CD133-cells. So we suggested that glioblastoma stem like cells were existed in CD133+ cells in U251 cell line with characteristics of self-renew and generation of an unlimited progeny of non-tumorigenic cells. Molecular and functional characterization of such a tumorigenic population may be exploited in the development of novel cancer therapeutic drugs. PMID:19232461

Qiang, Lei; Yang, Yong; Ma, Yan-Jun; Chen, Fei-Hong; Zhang, Ling-Bo; Liu, Wei; Qi, Qi; Lu, Na; Tao, Lei; Wang, Xiao-Tang; You, Qi-Dong; Guo, Qing-Long



Alu expression in human cell lines and their retrotranspositional potential  

PubMed Central

Background The vast majority of the 1.1 million Alu elements are retrotranspositionally inactive, where only a few loci referred to as ‘source elements’ can generate new Alu insertions. The first step in identifying the active Alu sources is to determine the loci transcribed by RNA polymerase III (pol III). Previous genome-wide analyses from normal and transformed cell lines identified multiple Alu loci occupied by pol III factors, making them candidate source elements. Findings Analysis of the data from these genome-wide studies determined that the majority of pol III-bound Alus belonged to the older subfamilies Alu S and Alu J, which varied between cell lines from 62.5% to 98.7% of the identified loci. The pol III-bound Alus were further scored for estimated retrotransposition potential (ERP) based on the absence or presence of selected sequence features associated with Alu retrotransposition capability. Our analyses indicate that most of the pol III-bound Alu loci candidates identified lack the sequence characteristics important for retrotransposition. Conclusions These data suggest that Alu expression likely varies by cell type, growth conditions and transformation state. This variation could extend to where the same cell lines in different laboratories present different Alu expression patterns. The vast majority of Alu loci potentially transcribed by RNA pol III lack important sequence features for retrotransposition and the majority of potentially active Alu loci in the genome (scored high ERP) belong to young Alu subfamilies. Our observations suggest that in an in vivo scenario, the contribution of Alu activity on somatic genetic damage may significantly vary between individuals and tissues.



A Comparative Analysis of Extra-Embryonic Endoderm Cell Lines  

PubMed Central

Prior to gastrulation in the mouse, all endodermal cells arise from the primitive endoderm of the blastocyst stage embryo. Primitive endoderm and its derivatives are generally referred to as extra-embryonic endoderm (ExEn) because the majority of these cells contribute to extra-embryonic lineages encompassing the visceral endoderm (VE) and the parietal endoderm (PE). During gastrulation, the definitive endoderm (DE) forms by ingression of cells from the epiblast. The DE comprises most of the cells of the gut and its accessory organs. Despite their different origins and fates, there is a surprising amount of overlap in marker expression between the ExEn and DE, making it difficult to distinguish between these cell types by marker analysis. This is significant for two main reasons. First, because endodermal organs, such as the liver and pancreas, play important physiological roles in adult animals, much experimental effort has been directed in recent years toward the establishment of protocols for the efficient derivation of endodermal cell types in vitro. Conversely, factors secreted by the VE play pivotal roles that cannot be attributed to the DE in early axis formation, heart formation and the patterning of the anterior nervous system. Thus, efforts in both of these areas have been hampered by a lack of markers that clearly distinguish between ExEn and DE. To further understand the ExEn we have undertaken a comparative analysis of three ExEn-like cell lines (END2, PYS2 and XEN). PYS2 cells are derived from embryonal carcinomas (EC) of 129 strain mice and have been characterized as parietal endoderm-like [1], END2 cells are derived from P19 ECs and described as visceral endoderm-like, while XEN cells are derived from blastocyst stage embryos and are described as primitive endoderm-like. Our analysis suggests that none of these cell lines represent a bona fide single in vivo lineage. Both PYS2 and XEN cells represent mixed populations expressing markers for several ExEn lineages. Conversely END2 cells, which were previously characterized as VE-like, fail to express many markers that are widely expressed in the VE, but instead express markers for only a subset of the VE, the anterior visceral endoderm. In addition END2 cells also express markers for the PE. We extended these observations with microarray analysis which was used to probe and refine previously published data sets of genes proposed to distinguish between DE and VE. Finally, genome-wide pathway analysis revealed that SMAD-independent TGFbeta signaling through a TAK1/p38/JNK or TAK1/NLK pathway may represent one mode of intracellular signaling shared by all three of these lines, and suggests that factors downstream of these pathways may mediate some functions of the ExEn. These studies represent the first step in the development of XEN cells as a powerful molecular genetic tool to study the endodermal signals that mediate the important developmental functions of the extra-embryonic endoderm. Our data refine our current knowledge of markers that distinguish various subtypes of endoderm. In addition, pathway analysis suggests that the ExEn may mediate some of its functions through a non-classical MAP Kinase signaling pathway downstream of TAK1.

Brown, Kemar; Legros, Stephanie; Artus, Jerome; Doss, Michael Xavier; Khanin, Raya; Hadjantonakis, Anna-Katerina; Foley, Ann



In vitro uptake of polystyrene microspheres: effect of particle size, cell line and cell density  

Microsoft Academic Search

Uptake of polycation–DNA particles is the first step in achieving gene delivery with non-viral vehicles. One of the important characteristics determining uptake of DNA particles is their size. Here we have characterized the ability of several cell lines to internalise labelled polystyrene microspheres of different sizes. All the cell lines tested ingested 20-nm microspheres avidly. With larger microspheres (93, 220,

Wolfgang Zauner; Neil A Farrow; Adrian M. R Haines



Aldehyde Dehydrogenase 1 Identifies Cells with Cancer Stem Cell-Like Properties in a Human Renal Cell Carcinoma Cell Line  

PubMed Central

Cancer stem cells (CSC) or cancer stem cell-like cells (CSC-LCs) have been identified in many malignant tumors. CSCs are proposed to be related with drug resistance, tumor recurrence, and metastasis and are considered as a new target for cancer treatment; however, there are only a few reports on CSCs or CSC-LCs in renal cell carcinoma (RCC). Different approaches have been reported for CSC identification, but there are no universal markers for CSC. We used two different approaches, the traditional side population (SP) approach, and the enzymatic (aldehyde dehydrogenase 1 (ALDH1)) approach to identify CSC-LC population in two RCC cell lines, ACHN and KRC/Y. We found that ACHN and KRC/Y contain 1.4% and 1.7% SP cells, respectively. ACHN SP cells showed a higher sphere forming ability, drug resistance, and a slightly higher tumorigenic ability in NOD/SCID mice than Non-SP (NSP) cells, suggesting that cells with CSC-LC properties are included in ACHN SP cells. KRC/Y SP and NSP cells showed no difference in such properties. ALDH1 activity analysis revealed that ACHN SP cells expressed a higher level of activity than NSP cells (SP vs. NSP: 32.7% vs 14.6%). Analysis of ALDH1-positive ACHN cells revealed that they have a higher sphere forming ability, self-renewal ability, tumorigenicity and express higher mRNA levels of CSC-LC property-related genes (e.g., ABC transporter genes, self-replication genes, anti-apoptosis genes, and so forth) than ALDH1-negative cells. Drug treatment or exposure to hypoxic condition induced a 2- to 3-fold increase in number of ALDH1-positive cells. In conclusion, the results suggest that the ALDH1-positive cell population rather than SP cells show CSC-LC properties in a RCC cell line, ACHN.

Ueda, Kosuke; Ogasawara, Sachiko; Akiba, Jun; Nakayama, Masamichi; Todoroki, Keita; Ueda, Keiko; Sanada, Sakiko; Suekane, Shigetaka; Noguchi, Masanori; Matsuoka, Kei; Yano, Hirohisa



Immunocytochemical analysis of human synovial lining cells: phenotypic relation to other marrow derived cells.  

PubMed Central

The antigenic phenotype of human synovial lining cells in normal and hyperplastic synovium intima was determined with a panel of monoclonal antibodies directed against a large number of well defined myeloid (macrophage/granulocyte associated) antigens. Synovial lining cells express numerous macrophage associated antigens, including CD11b (CR3), CD13, CD14, CD16 (FcRIII), CD18, CD32 (FcRII), CD45 (leucocyte common antigen), CD54 (ICAM-1), CD64 (FcRI), CD68, and CD71 (transferrin receptor). Few synovial lining cells expressed CD11a (LFA-1) and CD11c (p150,95). Subintimal macrophages expressed all the macrophage associated antigens which were present on synovial lining cells and, in addition, expressed CD15a, CD25 (interleukin-2 receptor), CD34, and CD35 (C3b receptor), none of which was present on synovial lining cells. Synovial lining cell expression of a wide range of macrophage antigens argues in favour of their marrow origin and membership of the mononuclear phagocyte system. Images

Athanasou, N A; Quinn, J



Establishment and characterisation of two cell lines derived from a primary adenocarcinoma of the duodenum  

PubMed Central

Aims—To establish two cell lines from a primary duodenal adenocarcinoma; to describe the morphological, growth, ploidy, and immunophenotypic characteristics of these cell lines. Methods—The cell lines, designated DAC/S and DAC/E, were characterised using both in vitro and in vivo cell culture techniques, light and electron microscopy, immunocytochemistry, and FACS analyses. Results—Both cell lines have an epithelial origin, are aneuploid and display characteristics of transformed cells. The cell lines differ from each other in morphology, doubling time and serum requirements. These cell lines are anchorage dependent and do not grow in nude mice. Conclusions—DAC/S and DAC/E cell lines are derived from neoplastic epithelium and could provide in vitro model systems for future investigations of the cell and molecular biology of duodenal neoplasia. Images

Golding, M; Stamp, G W H; Oates, T; Lalani, E-N



Production of skeletal muscle elements by cell lines derived from neoplastic rat mammary epithelial stem cells.  


Single-cell-cloned cell lines intermediate in morphology between the cuboidal epithelial and fully elongated myoepithelial-like cells have been isolated from the single-cell-cloned epithelial stem cell lines Rama 25 and Rama 37 originally obtained from dimethylbenz(a)anthracene-induced mammary tumors from Sprague-Dawley and Wistar-Furth rats, respectively. These are designated Rama 25-l1, Rama 25-l2, Rama 25-l4 (Sprague-Dawley) and Rama 50-55, Rama 59, and Rama 60 (Wistar-Furth), respectively. When growing as tumors in nude mice or syngeneic Wistar-Furth rats, respectively, many of the newly cloned cell lines give rise to spindle and giant, multinucleated cells which stain immunocytochemically with antisera to myoglobin and myosin and contain longitudinal fibrils, some of which contain phosphotungstic acid-hematoxylin-staining cross-striations. Ultrastructural analysis demonstrates the presence of A-, l-, and H-bands and Z-discs and the hexagonal arrangement of thick and thin filaments characteristic of skeletal muscle. Similar results are obtained with selected cloned cell lines growing on floating collagen gels in vitro. Thus, a developmentally committed mammary epithelial cell can give rise, under suitable conditions, to a well-differentiated mesenchymal lineage, that of skeletal muscle. It is suggested that such cells may be responsible for the generation of the well-differentiated mesenchymal elements seen in the mixed (epithelial and myoepithelial) tumors of glandular origin. PMID:6713401

Rudland, P S; Dunnington, D J; Gusterson, B; Monaghan, P; Hughes, C M



Adhesion of Actinobacillus actinomycetemcomitans to a human oral cell line.  


Two quantitative, rapid assays were developed to study the adhesion of Actinobacillus actinomycetemcomitans, an oral bacterium associated with periodontal disease, to human epithelial cells. The human oral carcinoma cell line KB was grown in microtiter plates, and adherent bacteria were detected by an enzyme-linked immunosorbent assay with purified anti-A. actinomycetemcomitans serum and horseradish peroxidase-conjugated secondary antibody or [3H]thymidine-labeled bacteria. Adhesion was found to be time dependent and increased linearly with increasing numbers of bacteria added. Variation in the level of adhesion was noted among strains of A. actinomycetemcomitans. Adhesion was not significantly altered by changes in pH (from pH 5 to 9) but was sensitive to sodium chloride concentrations greater than 0.15 M. Pooled human saliva was inhibitory for adhesion when bacteria were pretreated with saliva before being added to the cells. Pretreatment of the KB cells with saliva did not inhibit adhesion. Protease treatment of A. actinomycetemcomitans reduced adhesion of the bacteria to KB cells. The data are consistent with the hypothesis that a protein(s) is required for bacterial adhesion and that host components may play a role in modulating adhesion to epithelial cells. PMID:8063383

Mintz, K P; Fives-Taylor, P M



Cytotoxicity of cashew flavonoids towards malignant cell lines.  


The leaves of the Cashew plant (Anacardium occidentale L.) are used by the folk medicine in South America and West Africa. This plant is rich in flavonoids, which are polyphenolic compounds widespread in plants, and that have diverse physiological effects. In a sub-acute toxicity assay it was found that an ethanolic extract of Cashew leaves elicited lymphopenia in rats. The extract was also found to be cytotoxic and to induce apoptosis in Jurkat (acute lymphoblastic leukemia) cells. The crude ethanolic extract was fractionated and resolved by HPLC. One of the four fractions obtained led to the isolation of the biflavonoid agasthisflavone. [(3)H]-thymidine incorporation assays and flow cytometry analysis showed that the isolated compound displayed a high anti-proliferative effect in Jurkat cells with an IC(50) of 2.4 ?g/ml (4.45 ?M). The effect of agathisflavone on the acute promyelocytic leukemia cell line HL60, Burkitt lymphoma Raji cells and Hep-2 laryngeal carcinoma cells was also tested. The two latter ones were only mildly affected by agathisflavone. It is also shown that agathisflavone induces apoptosis in Jurkat cells and it this proposed that this is the likely mechanism of agathisflavone specific cytotoxicity. PMID:21106357

Konan, Nzi André; Lincopan, Nilton; Díaz, Ingrit Elida Collantes; de Fátima Jacysyn, Jacqueline; Tiba, Mirtes Midori Tanae; Amarante Mendes, João Gustavo Pessini; Bacchi, Elfriede Marianne; Spira, Beny



Characterization of a Dopaminergic Stimulatory Factor Derived from Monoclonal Cell Lines of Striatal Origin.  

National Technical Information Service (NTIS)

A lysate of an immortalized monoclonal cell line derived from the striatum (X61) contains two types of chemically distinct factors which are capable of increasing the dopamine content of an immortalized, dopaminergic mouse mesencephalic cell line (MN9D). ...

A. Heller L. Won M. Gross



Verification and Unmasking of Widely Used Human Esophageal Adenocarcinoma Cell Lines  

PubMed Central

For decades, hundreds of different human tumor type–specific cell lines have been used in experimental cancer research as models for their respective tumors. The veracity of experimental results for a specific tumor type relies on the correct derivation of the cell line. In a worldwide effort, we verified the authenticity of all available esophageal adenocarcinoma (EAC) cell lines. We proved that the frequently used cell lines SEG-1 and BIC-1 and the SK-GT-5 cell line are in fact cell lines from other tumor types. Experimental results based on these contaminated cell lines have led to ongoing clinical trials recruiting EAC patients, to more than 100 scientific publications, and to at least three National Institutes of Health cancer research grants and 11 US patents, which emphasizes the importance of our findings. Widespread use of contaminated cell lines threatens the development of treatment strategies for EAC.

Boonstra, Jurjen J.; van Marion, Ronald; Beer, David G.; Lin, Lin; Chaves, Paula; Ribeiro, Catarina; Pereira, A. Dias; Roque, Lucia; Darnton, S. Jane; Altorki, Nasser K.; Schrump, David S.; Klimstra, David S.; Tang, Laura H.; Eshleman, James R.; Alvarez, Hector; Shimada, Yutaka; van Dekken, Herman; Tilanus, Hugo W.



Raman spectra and discrimination of NPC cell line CNE1 and normal nasopharyngeal cell line NP69  

NASA Astrophysics Data System (ADS)

As a non-destructive and non-invasive technique, Raman spectroscopy (RS) plays an important role in the field of biomedical research. Great progress has been made in the research of biological samples from cellular level to macro-tissues. In this letter, advances of RS in tumor cells and some statistic algorithm developed in recent years for cancer differentiation and diagnosis are introduced. Also, Raman spectra of Nasopharyngeal Carcinoma (NPC) cell line CNE1 and normal nasopharyngeal cell line NP69 are acquired by confocal Raman micro-spectroscopy system. Raman bands are analyzed and compared to investigate the differences and relationship between CNE1 and NP69, Principle Components Analysis (PCA) is used to classify CNE1 and NP69 accurately with an accuracy of 100%. Comparing with CNE1, a blue-shift is observed in NP69 cells at band 936cm-1 and 2935cm-1 which are assigned to C-C stretch and CH3 stretching, respectively. Meanwhile, a red-shift is observed at 1338cm-1 assigned to A, G and C-H deformation vibration of protein. The results show that Raman spectroscopy has its potential and reliability to be one of the diagnostic methods for NPC and at the same time can provide valuable information for cancer early diagnosis.

Chen, Yang; Li, Yong-Zeng; Su, Ying; Lin, Ju-Qiang; Pan, Jian-Ji; Chen, Rong; Zou, Chang-Yan; Lin, Shaojun; Li, Chao



Evaluation of TV cell line viral susceptibility using conventional cell culture techniques  

Microsoft Academic Search

Despite the fact that a lot of methods have been developed for rapid virus detection, classic cell culture is still “the golden\\u000a standard”. The range of viruses that can be isolated and cultured in cell line systems is often limited by the susceptibility\\u000a of cells to support viral replication. Since the primary cell culture, the best cellular system available to

Coralia Bleotu; Carmen C. Diaconu; Mihaela Chivu; Irina Alexiu; Simona M. Ruta; Costin Cernescu



New cell lines from mouse epiblast share defining features with human embryonic stem cells  

Microsoft Academic Search

The application of human embryonic stem (ES) cells in medicine andbiologyhasaninherentrelianceonunderstandingthestarting cellpopulation.HumanEScellsdifferfrommouseEScellsandthe specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo before implantation in the uterus1-5. Here we show that cell lines can be derived from the epiblast, a tissue of the post- implantation embryo that

Josh G. Chenoweth; Frances A. Brook; Timothy J. Davies; Edward P. Evans; David L. Mack; Richard L. Gardner; Paul J. Tesar; Ronald D. G. McKay



Stable cell line of T-SV40 immortalized human glomerular visceral epithelial cells  

Microsoft Academic Search

Stable cell line of T-SV40 immortalized human glomerular visceral epithelial cells. Human subcultures (third passage) of glomerular visceral epithelial cells (VEC) isolated from one month old kidney were successfully transfected by two recombinant plasmids containing the cloned oncogenes from the simian virus 40 large T antigen and H-ras gene. One postcrisis cell clone (56\\/10 A1) was selected, propagated and characterized.

Françoise Delarue; Angela Virone; Jacqueline Hagege; Roger Lacave; Marie-Nëlle Peraldi; Colette Adida; Eric Rondeau; Jean Feunteun; Jean-Daniel Sraer



Selenium retention and inhibition of cell growth in mouse mammary epithelial cell lines in vitro  

Microsoft Academic Search

The steady state levels of growth inhibitory doses of inorganic selenium were examined in five different mammary epithelial\\u000a cell lines: MOD, COMMA-D, COMMA-F, COMMA-T, and YN-4. The retention of selenium was monitored using a radioactive isotope,75Se. Growth inhibition correlated with high levels of selenium in the cell. Generally, the retention of intracellular selenium\\u000a was not dependent upon cell density, cell

Daniel Medina; David Morrison; Carol J. Oborn



Reference Maps of Human ES and iPS Cell Variation Enable High-Throughput Characterization of Pluripotent Cell Lines  

PubMed Central

SUMMARY The developmental potential of human pluripotent stem cells suggests that they can produce disease-relevant cell types for biomedical research. However, substantial variation has been reported among pluripotent cell lines, which could affect their utility and clinical safety. Such cell-line-specific differences must be better understood before one can confidently use embryonic stem (ES) or induced pluripotent stem (iPS) cells in translational research. Toward this goal we have established genome-wide reference maps of DNA methylation and gene expression for 20 previously derived human ES lines and 12 human iPS cell lines, and we have measured the in vitro differentiation propensity of these cell lines. This resource enabled us to assess the epigenetic and transcriptional similarity of ES and iPS cells and to predict the differentiation efficiency of individual cell lines. The combination of assays yields a scorecard for quick and comprehensive characterization of pluripotent cell lines.

Bock, Christoph; Kiskinis, Evangelos; Verstappen, Griet; Gu, Hongcang; Boulting, Gabriella; Smith, Zachary D.; Ziller, Michael; Croft, Gist F.; Amoroso, Mackenzie W.; Oakley, Derek H.; Gnirke, Andreas; Eggan, Kevin; Meissner, Alexander



Establishment and characterisation of cell lines from patients with lung cancer (predominantly small cell carcinoma).  

PubMed Central

Tissue samples from 59 patients with lung cancer have been used to establish cell lines in culture. The primary diagnosis was small cell carcinoma in all except four. Most of the samples were of bone marrow but pleural effusions, lymph node biopsies and skin metastases were also included. The samples were usually split between HITES serum-free medium and HITES plus 2.5% foetal calf serum. A total of 19 cell lines were established and characterised. One line is large cell anaplastic lung carcinoma, four are B-lymphoblastoid and fourteen are small cell lung cancer. Considerable heterogeneity in gross morphology, neuroendocrine differentiation (by electron microscopy) and content of the enzyme L-dopa decarboxylase was seen. The use of HITES plus 2.5% foetal calf serum resulted in better establishment of cultures than did serum-free HITES.

Baillie-Johnson, H.; Twentyman, P. R.; Fox, N. E.; Walls, G. A.; Workman, P.; Watson, J. V.; Johnson, N.; Reeve, J. G.; Bleehen, N. M.



Establishment and maintenance of three Chinese human embryonic stem cell lines  

Microsoft Academic Search

To establish a potential resource for cell therapy and a developmental model for human diseases, we had isolated three Chinese\\u000a human embryonic stem cell lines from the inner cell mass of human blastocysts in 2002. All the three cell lines were grown\\u000a on mouse embryonic fibroblasts as feeder cells; one of these cell lines, chHES-3, has maintained its normal karyotype

Di Zhou; Ge Lin; Chang-Qing Xie; Bo Xiong; Xiao-Ying Zhou; Xiao-Bing Qian; Yi Liao; Guang-Xiu Lu



Retinal ganglion cell line apoptosis induced by hydrostatic pressure.  


Cellular responses to changes in pressure are implicated in numerous disease processes. In glaucoma apoptosis of retinal ganglion cells (RGCs) is associated with elevated intra-ocular pressure, however, the exact cellular mechanisms remain unclear. We have previously shown that pressure can induce apoptosis in B35 and PC12 neuronal cell lines, using an in vitro model for pressure elevation. A novel RGC line allows us to study the effects of pressure on retinal neurons. 'RGC-5' cultures were subjected to elevated ambient hydrostatic pressure conditions in our model. Experimental pressure conditions were 100 mm Hg and 30 mm Hg, representing acute (high) and chronic (lower-pressure) glaucoma, and 15 mm Hg for normal intra-ocular pressure, set above atmospheric pressure for 2 h. Negative controls were treated identically except for the application of pressure, while positive controls were generated by treatment with a known apoptotic stimulus. Apoptosis was determined by a combination of cell morphology and specific TUNEL and Annexin V fluorescent markers. These were assessed simultaneously by laser scanning cytometry (LSC), which also enabled quantitative marker analysis. RGC-5 neurons showed a significantly increased proportion of apoptotic cells compared with controls; maximal at 100 mm Hg, moderate at 30 mm Hg and not statistically significant at 15 mm Hg. This graded response, proportionate to the level of pressure elevation, is representative of the severity of analogous clinical settings (acute, chronic glaucoma and normal). These results complement earlier findings of pressure-induced apoptosis in other neuronal cultures. They suggest the possibility of novel mechanisms of pressure-related mechanotransduction and cell death, relevant to the pathogenesis of diseases such as glaucoma. PMID:16638612

Agar, Ashish; Li, Shaojuan; Agarwal, Neeraj; Coroneo, Minas T; Hill, Mark A



Multidrug resistance in tumour cells: characterization of the multidrug resistant cell line K562-Lucena 1.  


Multidrug resistance to chemotherapy is a major obstacle in the treatment of cancer patients. The best characterised mechanism responsible for multidrug resistance involves the expression of the MDR-1 gene product, P-glycoprotein. However, the resistance process is multifactorial. Studies of multidrug resistance mechanisms have relied on the analysis of cancer cell lines that have been selected and present cross-reactivity to a broad range of anticancer agents. This work characterises a multidrug resistant cell line, originally selected for resistance to the Vinca alkaloid vincristine and derived from the human erythroleukaemia cell K562. This cell line, named Lucena 1, overexpresses P-glycoprotein and have its resistance reversed by the chemosensitisers verapamil, trifluoperazine and cyclosporins A, D and G. Furthermore, we demonstrated that methylene blue was capable of partially reversing the resistance in this cell line. On the contrary, the use of 5-fluorouracil increased the resistance of Lucena 1. In addition to chemotherapics, Lucena 1 cells were resistant to ultraviolet A radiation and hydrogen peroxide and failed to mobilise intracellular calcium when thapsigargin was used. Changes in the cytoskeleton of this cell line were also observed. PMID:11246270

Rumjanek, V M; Trindade, G S; Wagner-Souza, K; de-Oliveira, M C; Marques-Santos, L F; Maia, R C; Capella, M A



Establishment of Immortalized Human Erythroid Progenitor Cell Lines Able to Produce Enucleated Red Blood Cells  

PubMed Central

Transfusion of red blood cells (RBCs) is a standard and indispensable therapy in current clinical practice. In vitro production of RBCs offers a potential means to overcome a shortage of transfusable RBCs in some clinical situations and also to provide a source of cells free from possible infection or contamination by microorganisms. Thus, in vitro production of RBCs may become a standard procedure in the future. We previously reported the successful establishment of immortalized mouse erythroid progenitor cell lines that were able to produce mature RBCs very efficiently. Here, we have developed a reliable protocol for establishing immortalized human erythroid progenitor cell lines that are able to produce enucleated RBCs. These immortalized cell lines produce functional hemoglobin and express erythroid-specific markers, and these markers are upregulated following induction of differentiation in vitro. Most importantly, these immortalized cell lines all produce enucleated RBCs after induction of differentiation in vitro, although the efficiency of producing enucleated RBCs remains to be improved further. To the best of our knowledge, this is the first demonstration of the feasibility of using immortalized human erythroid progenitor cell lines as an ex vivo source for production of enucleated RBCs.

Kurita, Ryo; Suda, Noriko; Sudo, Kazuhiro; Miharada, Kenichi; Hiroyama, Takashi; Miyoshi, Hiroyuki; Tani, Kenzaburo; Nakamura, Yukio



Selection of cell lines with enhanced invasive phenotype from xenografts of the human prostate cancer cell line WPE1-NB26.  


Prostate cancer is a leading cause of death from cancer in American men and metastasis the main cause of death. To better understand the disease and accelerate development of new therapies, in vivo models that reflect different disease stages are needed. A family of cell lines that mimics multiple steps in cancer development and tumor progression has been developed in our laboratory from the parent, non-tumorigenic, RWPE-1 cell line by transformation with N-methyl-N-nitrosourea (MNU). The MNU cell lines mimic multiple steps in tumor progression where WPE1-NB26 is the most malignant cell line. WPE1-NB26 cells form metastases in the lungs of athymic, male, nude mice after intravenous injection. Two new cell lines, WPE1-NB26-64 and WPE1-NB26-65, showing more malignant characteristics than the parent WPE1-NB26 cell line, were derived from tumors after subcutaneous injection of WPE1-NB26 cells into nude mice. The WPE1-NB26-64 and WPE1-NB26-65 cell lines show an increase in anchorage-dependent growth and invasive ability as compared to the parent WPE1-NB26 cells. While the parent WPE1-NB26 cells express barely detectable levels, the new cell lines produce high levels of matrix metalloproteinase MMP-2 and detectable levels of MMP-9. By immunostaining, all three cell lines were positive for cytokeratins CK18 and CK5/14. These cell lines, having the same lineage, represent additional steps in the multi-step process of tumor progression and provide novel and useful cell models for studies on tumor progression and for drug development for the treatment of prostate cancer. PMID:16471037

Rivette, Amanda S; Tokar, Erik J; Williams, Daniel E; Mackenzie, Charles D; Ablin, Richard J; Webber, Mukta M



Erythropoietin modulates intracellular calcium in a human neuroblastoma cell line  

PubMed Central

Recent investigations have shown that the glycoprotein erythropoietin (Epo) and its specific receptor (EpoR) are present in the mammalian brain including human, monkey and mouse. These findings suggest a local action of Epo in the nervous system. The aim of this study was to elucidate a possible functional interaction of Epo with neuronal cells. To examine the influence of externally applied Epo on Ca2+ homeostasis the human neuroblastoma cell line SK-N-MC was chosen as a suitable in vitro model for undifferentiated neuronal cells. Expression of the EpoR in SK-N-MC cells was detected by reverse transcription-PCR, Western blot and immunofluorescence analysis. Patch-clamp studies of SK-N-MC cells confirmed the expression of T-type Ca2+ channels, whose peak macroscopic current was increased by the addition of recombinant human Epo (rhEpo) to the bathing medium. Confocal laser scanning microscopy analysis of SK-N-MC cells confirmed a transient increase in intracellular free [Ca2+] in response to externally applied rhEpo. The transient response to Epo was dependent on external Ca2+ and remained even after depletion of internal Ca2+ stores by caffeine or thapsigargin. However, after depletion the response to Epo was absent when cells were superfused with the T-type Ca2+ channel blocker flunarizine. This study demonstrates that Epo can interact with neuronal cells by affecting Ca2+ homeostasis through an increase in Ca2+ influx via plasma membrane T-type voltage-dependent Ca2+ channels.

Assandri, Roberta; Egger, Marcel; Gassmann, Max; Niggli, Ernst; Bauer, Christian; Forster, Ian; Gorlach, Agnes



Differential induction of numerical chromosome changes by sodium butyrate in two transformed cell lines  

Microsoft Academic Search

Transformed cell lines generally exhibit similar response to a given chemical. However, when two transformed cell lines, the Chinese hamster (CHO 9) and mouse (L929) were treated with sodium butyrate (SB) to investigate its effect on chromosome distribution, the two lines behaved differently from each other. At concentrations as low as 10?7 M, the CHO 9 cells exhibited polyploidy as

Patricia Gomez-Vargas; Baldev K Vig



Inhibition of NF-?B Activity Enhances TRAIL Mediated Apoptosis in Breast Cancer Cell Lines  

Microsoft Academic Search

Most breast cancer cell lines are resistant to TNF-related apoptosis inducing ligand (TRAIL) induced apoptosis. In sensitive breast cancer cell lines TRAIL rapidly induces the cleavage and activation of caspases leading to the subsequent cleavage of downstream caspase substrates. In contrast, there is no caspase activation in the resistant cell lines. The transcription factor NF-?B can inhibit apoptosis induced by

Maccon M. Keane; Yaffa Rubinstein; Mauricio Cuello; Seth A. Ettenberg; Priya Banerjee; Marion M. Nau; Stan Lipkowitz



Establishment and Characterization of Seven Continuous Cell Lines from Freshwater Fish  

Microsoft Academic Search

Seven continuous cell lines were established from salmonid and nonsalmonid fishes. Salmonid cell lines derived from rainbow trout Oncorhynchus mykiss and chum salmon O. keta were designated RTE and RTE-2 (rainbow trout embryo), RTT (rainbow trout tail), and SEH (“sake” or chum salmon embryo head). Nonsalmonid cell lines derived from pond smelt Hypomesus olidus, chevron snakehead Channa striata, and goldfish

R. D. Fernandez; M. Yoshimizu; T. Kimura; Y. Ezura



Chemical Data Mining of the NCI Human Tumor Cell Line Database  

Microsoft Academic Search

The NCI Developmental Therapeutics Program Human Tumor cell line Dataset is a publicly available database that contains cellular assay screening data for over 40,000 compounds tested in sixty human tumor cell lines. The database also contains microarray assay gene expression data for the cell lines, and so it provides an excellent information resource particularly for testing data mining methods that

Huijun Wang; Jonathan Klinginsmith; Xiao Dong; Adam C. Lee; Rajarshi Guha; Yuqing Wu; Gordon M. Crippen; David J. Wild



Identification of cancer stem cells from hepatocellular carcinoma cell lines and their related microRNAs.  


The aim of this study was to identify cancer stem cells (CSC) from three hepatocellular carcinoma (HCC) cell lines and to screen for specific microRNAs (miRNAs) regulating CSCs. Side population (SP) phenotype analysis was used. Four factors in the staining process, the incubation time, shaking interval, culture time and Hoechst 33342 concentration were explored, respectively, to define the SP subtype. CSC characteristics of SP cells were verified by sphere-forming assay and tumorigenic ability in NOD/SCID mice. QPCR assay for 370 miRNAs was performed to identify the differential miRNA expression between SP and Non-SP (NSP) cells in the PLC/PRF/5 cell line. The selected miRNAs were tested again in SP and NSP cells from Huh-7 and Hep-3B cell lines by qPCR assay. All four factors influenced SP percentage, when the other three conditions were fixed, the optimal Hoechst 33342 concentrations determined were 11 µg/ml for PLC/PRF/5 cells, 4 µg/ml for Huh-7 and 5 µg/ml for Hep-3B cells. The resultant SP percentage was 0.73±0.12%, 0.49±0.04% and 0.63±0.08%, respectively. The purity of sorted SP cells was >85%. Floating spheres were formed by SP cells from all three cell lines, while NSP cells did not form a single floating sphere. Mice injected with SP cells on the right side formed more tumor masses compared to their counterpart NSP at the same injection dosage; qPCR profiling identified 27 differentially expressed miRNAs in PLC/PRF/5 cells. Subsequent qPCR assay showed that miR-9* and miR-194 were also downregulated in SP cells from Huh-7 and Hep-3B. The present study identified CSCs via SP and sphere-forming assay from three liver cancer cell lines. Altogether, 27 CSC-specific miRNAs were determined in PLC/PRF/5; miR-9* and miR-194 were identified as the common CSC-specific miRNAs across the three HCC cell lines. PMID:24002436

Xu, Yangmei; Xie, Yunqing; Wang, Xiangru; Chen, Xuefang; Liu, Qingyin; Ying, Mingang; Zheng, Qiuhong



Two new protocols to enhance the production and isolation of human induced pluripotent stem cell lines  

Microsoft Academic Search

There are two critical stages in the retroviral reprogramming of somatic cells to produce human induced pluripotent stem cell (hiPSC) lines. One is the production of high titer virus required to reprogram somatic cells; the other is identification of true hiPSC colonies from heterogeneous cell populations, and their isolation and expansion to generate a sustainable, pluripotent stem cell line. Here

Emily Dick; Elena Matsa; Jayson Bispham; Mojgan Reza; Michela Guglieri; Andrew Staniforth; Sue Watson; Rajendra Kumari; Hanns Lochmüller; Lorraine Young; David Darling; Chris Denning



Oxidative stress induces hypomethylation of LINE-1 and hypermethylation of the RUNX3 promoter in a bladder cancer cell line.  


Increased oxidative stress and changes in DNA methylation are frequently detected in bladder cancer patients. We previously demonstrated a relationship between increased oxidative stress and hypomethylation of the transposable long-interspersed nuclear element-1 (LINE-1). Promoter hypermethylation of a tumor suppressor gene, runt-related transcription factor 3 (RUNX3), may also be associated with bladder cancer genesis. In this study, we investigated changes of DNA methylation in LINE-1 and RUNX3 promoter in a bladder cancer cell (UM-UC-3) under oxidative stress conditions, stimulated by challenge with H2O2 for 72 h. Cells were pretreated with an antioxidant, tocopheryl acetate for 1 h to attenuate oxidative stress. Methylation levels of LINE-1 and RUNX3 promoter were measured by combined bisulfite restriction analysis PCR and methylation-specific PCR, respectively. Levels of LINE-1 methylation were significantly decreased in H2O2-treated cells, and reestablished after pretreated with tocopheryl acetate. Methylation of RUNX3 promoter was significantly increased in cells exposed to H2O2. In tocopheryl acetate pretreated cells, it was markedly decreased. In conclusion, hypomethylation of LINE-1 and hypermethylation of RUNX3 promoter in bladder cancer cell line was experimentally induced by reactive oxygen species (ROS). The present findings support the hypothesis that oxidative stress promotes urothelial cell carcinogenesis through modulation of DNA methylation. Our data also imply that mechanistic pathways of ROS-induced alteration of DNA methylation in a repetitive DNA element and a gene promoter might differ. PMID:23886181

Wongpaiboonwattana, Wikrom; Tosukhowong, Piyaratana; Dissayabutra, Thasinas; Mutirangura, Apiwat; Boonla, Chanchai



Doxycycline Alters Metabolism and Proliferation of Human Cell Lines  

PubMed Central

The tetracycline antibiotics are widely used in biomedical research as mediators of inducible gene expression systems. Despite many known effects of tetracyclines on mammalian cells–including inhibition of the mitochondrial ribosome–there have been few reports on potential off-target effects at concentrations commonly used in inducible systems. Here, we report that in human cell lines, commonly used concentrations of doxycycline change gene expression patterns and concomitantly shift metabolism towards a more glycolytic phenotype, evidenced by increased lactate secretion and reduced oxygen consumption. We also show that these concentrations are sufficient to slow proliferation. These findings suggest that researchers using doxycycline in inducible expression systems should design appropriate controls to account for potential confounding effects of the drug on cellular metabolism.

Cass, Ashley; Braas, Daniel; York, Autumn G.; Bensinger, Steven J.; Graeber, Thomas G.; Christofk, Heather R.



Doxycycline alters metabolism and proliferation of human cell lines.  


The tetracycline antibiotics are widely used in biomedical research as mediators of inducible gene expression systems. Despite many known effects of tetracyclines on mammalian cells-including inhibition of the mitochondrial ribosome-there have been few reports on potential off-target effects at concentrations commonly used in inducible systems. Here, we report that in human cell lines, commonly used concentrations of doxycycline change gene expression patterns and concomitantly shift metabolism towards a more glycolytic phenotype, evidenced by increased lactate secretion and reduced oxygen consumption. We also show that these concentrations are sufficient to slow proliferation. These findings suggest that researchers using doxycycline in inducible expression systems should design appropriate controls to account for potential confounding effects of the drug on cellular metabolism. PMID:23741339

Ahler, Ethan; Sullivan, William J; Cass, Ashley; Braas, Daniel; York, Autumn G; Bensinger, Steven J; Graeber, Thomas G; Christofk, Heather R



Cell lines derived from feline fibrosarcoma display unstable chromosomal aneuploidy and additionally centrosome number aberrations.  


The purpose of the study was to evaluate clonality and presence of numerical chromosomal and centrosomal aberrations in 5 established feline fibrosarcoma cell lines and in a fetal dermal fibroblast cell line as a control. The clonality of all cell lines was examined using limited-dilution cloning. The number of chromosomes was counted in metaphase spreads. The immunocytochemical analysis of centrosome numbers was performed by indirect immunofluorescence using a monoclonal antibody that targets ?-tubulin, a well-characterized component of centrosomes. Monoclonal cell populations could be established from all cell lines. In all feline fibrosarcoma cell lines, the number of chromosomes deviated abnormally from the normal feline chromosome number of 2n = 38, ranging from 19 to 155 chromosomes per cell. Centrosome hyperamplification was observed in all 5 feline fibrosarcoma cell lines with a proportion of cells (5.7 to 15.2%) having more than 2 centrosomes. In the control cell line, only 0.6% of the cells had more than 2 centrosomes. In conclusion, the examinations revealed that centrosome hyperamplification occurs in feline fibrosarcoma cell lines. The feline fibrosarcoma cell lines possessed 10 to 25 times as many cells with centrosome hyperamplification as the control cell line. These observations suggest an association of numerical centrosome aberrations with karyotype instability by increasing the frequency of chromosome missegregation. The results of this study may be helpful for further characterization of feline fibrosarcomas and may contribute to the knowledge of cytogenetic factors that may be important for the pathogenesis of feline fibrosarcomas. PMID:21527782

von Erichsen, J; Hecht, W; Löhberg-Gruene, C; Reinacher, M



Kaposi's sarcoma-derived cell line SLK is not of endothelial origin, but is a contaminant from a known renal carcinoma cell line.  


Kaposi's sarcoma (KS) is an endothelial cell-derived tumor. Investigations of the molecular mechanisms of KS pathogenesis and the identification of drugs for treatment of KS depend critically on valid cell-culture models. Two major immortalized cell lines are available for KS research. Recently, the KS cell line KS Y-1 has been shown to be cross-contaminated with the T24 urinary bladder cancer cell line (ATCC HTB-4). Here, we show by short tandem repeat profiling that the second KS cell line, SLK, is indistinguishable from the clear-cell renal-cell carcinoma cell line Caki-1. Immunocytochemical detection of cytokeratin expression confirmed the epithelial-cell origin of SLK cells. Our findings indicate that SLK cells are not of endothelial origin and should not be used in future studies as a model for KS-derived endothelial tumor cells. We suggest that in the future, more attention needs to be paid to the authenticity of cells in lines derived from human tissues. PMID:22987579

Stürzl, Michael; Gaus, Dominika; Dirks, Wilhelm G; Ganem, Don; Jochmann, Ramona



Characterisation and manipulation of docetaxel resistant prostate cancer cell lines  

PubMed Central

Background There is no effective treatment strategy for advanced castration-resistant prostate cancer. Although Docetaxel (Taxotere®) represents the most active chemotherapeutic agent it only gives a modest survival advantage with most patients eventually progressing because of inherent or acquired drug resistance. The aims of this study were to further investigate the mechanisms of resistance to Docetaxel. Three Docetaxel resistant sub-lines were generated and confirmed to be resistant to the apoptotic and anti-proliferative effects of increasing concentrations of Docetaxel. Results The resistant DU-145 R and 22RV1 R had expression of P-glycoprotein and its inhibition with Elacridar partially and totally reversed the resistant phenotype in the two cell lines respectively, which was not seen in the PC-3 resistant sublines. Resistance was also not mediated in the PC-3 cells by cellular senescence or autophagy but multiple changes in pro- and anti-apoptotic genes and proteins were demonstrated. Even though there were lower basal levels of NF-?B activity in the PC-3 D12 cells compared to the Parental PC-3, docetaxel induced higher NF-?B activity and I?B phosphorylation at 3 and 6 hours with only minor changes in the DU-145 cells. Inhibition of NF-?B with the BAY 11-7082 inhibitor reversed the resistance to Docetaxel. Conclusion This study confirms that multiple mechanisms contribute to Docetaxel resistance and the central transcription factor NF-?B plays an immensely important role in determining docetaxel-resistance which may represent an appropriate therapeutic target.



In vitro evaluation of a new nitrosourea, TCNU, against human small cell lung cancer cell lines  

Microsoft Academic Search

The cytotoxic activity of a new nitrosourea, TCNU, was compared with that of BCNU in five human small cell lung cancer cell lines in vitro. TCNU was found to be equivalent or inferior to BCNU when compared on a microgram to microgram basis. If the potential of in vitro phase II trials for selection of new drugs can be validated,

Henrik Roed; Lars L. Vindeløv; Mogens Spang-Thomsen; Ib J. Christensen; Heine Høi Hansen



Expression of Germ Cell Nuclear Factor (GCNF) by Ovarian Cancer Cell Lines  

Microsoft Academic Search

Ovarian cancer is the fifth most common cancer (neoplasm) among women and the third most common gynecological cancer behind endometrial and cervical cancer (1). Epithelial ovarian cancer (EOC) represents ~90% of all ovarian cancers. Recently, we have observed the novel expression of the nuclear receptor, the germ cell nuclear factor (GCNF) in several ovarian cancer cell lines. This is noteworthy

Sowmya Srikanthan; Jeffrey V. May


3-Bromopyruvate induces necrotic cell death in sensitive melanoma cell lines.  


Clinicians successfully utilize high uptake of radiolabeled glucose via PET scanning to localize metastases in melanoma patients. To take advantage of this altered metabolome, 3-bromopyruvate (BrPA) was used to overcome the notorious resistance of melanoma to cell death. Using four melanoma cell lines, BrPA triggered caspase independent necrosis in two lines, whilst the other two lines were resistant to killing. Mechanistically, sensitive cells differed from resistant cells by; constitutively lower levels of glutathione, reduction of glutathione by BrPA only in sensitive cells; increased superoxide anion reactive oxygen species, loss of outer mitochondrial membrane permeability, and rapid ATP depletion. Sensitive cell killing was blocked by N-acetylcysteine or glutathione. When glutathione levels were reduced in resistant cell lines, they became sensitive to killing by BrPA. Taken together, these results identify a metabolic-based Achilles' heel in melanoma cells to be exploited by use of BrPA. Future pre-clinical and clinical trials are warranted to translate these results into improved patient care for individuals suffering from metastatic melanoma. PMID:20430010

Qin, J-Z; Xin, H; Nickoloff, B J



Cytotoxic effects of mistletoe (Viscum album L.) in head and neck squamous cell carcinoma cell lines.  


Head and neck squamous cell carcinoma is a complex disease with several etiologic factors and different molecular changes that may trigger certain events; it is also globally one of the most common malignancies in this topography. Extracts from Viscum album L. (VA) (mistletoe) have been used as adjuvant therapies with promising results in several types of cancer, mainly in European countries. In vitro studies have demonstrated that various types of VA may have cytotoxicity in carcinoma cells, activating the apoptotic cascade or leading cells to necrosis. This study aimed to verify the effects of three types of VA extracts (Iscador Qu Spezial, Iscador P and Iscador M) in squamous cell carcinoma of the tongue cell lines SCC9 and SCC25, not previously studied. A concentration of 0.3 mg/ml (IC50) of the drugs induced apoptosis, affecting gene expression and protein levels of AKT, PTEN and CYCLIN D1. It was concluded that VA extracts have a cytotoxic effect on SCC9 and SCC25 cell lines, but while SCC9 cell line was more resistant to the action of the drugs, Iscador Qu Spezial and Iscador M have higher cytotoxic potential in both cell lines compared to Iscador P. PMID:24026291

Klingbeil, M Fátima G; Xavier, Flávia C A; Sardinha, Luiz R; Severino, Patricia; Mathor, Monica B; Rodrigues, Rodrigo V; Pinto, Décio S



Responses in mantle cell lymphoma cells to SNS-032 depend on the biological context of each cell line  

PubMed Central

SNS-032 is a potent inhibitor of cyclin-dependent kinases (Cdk) 2, 7 and 9 that regulate the cell cycle and transcription. Our studies in indolent primary chronic lymphocytic leukemia cells demonstrated that SNS-032 inhibited transcription, diminished the anti-apoptotic protein Mcl-1, and induced apoptosis. The present study focuses on evaluating this compound in four proliferating mantle cell lymphoma (MCL) lines (Jeko-1, Granta 519, Mino and SP-53). Consistent with its action against Cdk9 and Cdk7, SNS-032 inhibited the phosphorylation of RNA pol II in all 4 lines and blocked RNA synthesis. The transcripts and protein levels of short-lived proteins decreased, including cyclin D1 and Mcl-1. Cell growth was inhibited in a concentration-dependent manner in all lines. Apoptosis was induced in JeKo-1, Mino and SP-53 cells without disrupting cell cycle distribution. However, apoptosis was limited in Granta cells; rather, there was a significant reduction of clonogenic survival. SiRNA was used to specifically knock down Mcl-1 and cyclin D1 in JeKo-1 and Granta cells. Knocking down Mcl-1 induced significant apoptosis in Jeko-1 cells but not Granta cells. Reducing cyclin D1, rather than Mcl-1 was associated with loss of clonogenic survival in Granta cells. Thus, these results indicated that MCL cell lines have distinct mechanisms sustaining their survival, and the mechanism of action SNS-032 is dependent on the biological context of an individual line.

Chen, Rong; Chubb, Sherri; Cheng, Tiewei; Hawtin, Rachael E; Gandhi, Varsha; Plunkett, William



Cloning of human XAF1 gene promoter and assay of its transcription activity in a variety of cell lines  

Microsoft Academic Search

To investigate the regulation of tumor suppressor XAF1 gene expression in digestive system cancers, we studied XAF1 gene promoter\\u000a transcription activity and mRNA level in digestive system cancer cell lines (human hepatoma cell line HepG2, human colon cancer\\u000a cell line LoVo, and human gastric cancer cell line AGS) and nontumor cell lines (human embryonic liver cell line L02 (L02\\u000a cells)

Qiong Chen; Qing Yu; Yuhu Song; Peiyuan Li; Ying Chang; Zhijun Wang; Lifeng Liu; Wei Wu; Jusheng Lin



LCC15MB Cells are MDAMB-435: A Review of Misidentified Breast and prostate cell lines  

Microsoft Academic Search

Current stocks of the LCC15-MB cell line, which we originally isolated from a human breast-bone metastasis, were found to\\u000a be genetically matched to the MDA-MB-435 cell line from the Lombardi Cancer Center (MDA-MB-435-LCC) using comparative genomic\\u000a hybridisation, DNA microsatellite analysis and chromosomal number. LCC15-MB stocks used for our previously published studies\\u000a as well as the earliest available LCC15-MB cells also

Erik W. Thompson; Mark Waltham; Susan J. Ramus; Anne-Marie Hutchins; Jane E. Armes; Ian G. Campbell; Elizabeth D. Williams; Phillip R. Thompson; James M. Rae; Michael D. Johnson; Robert Clarke



All-trans-retinoic acid induces cell growth arrest in a human medulloblastoma cell line.  


Medulloblastomas (MBs) are the most common malignant brain tumors of childhood. Antitumor agents promoting long-term survival with limited toxicities are thus far lacking. Preliminary findings suggest that retinoic acid (RA) derivatives (retinoids) exert antitumor effects by inhibiting cell proliferation and inducing cell differentiation, apoptosis, and growth arrest, and RAs have been specifically shown to induce apoptosis in some MB cells. However, there is no conclusive evidence of retinoids inducing cell growth arrest in MBs. The aim of this study is to investigate whether retinoids play a role in cell-cycle arrest of MB cells. All-trans-retinoic acid (ATRA) was selected for these studies as it is known to have the capacity of inducing cell cycle arrest and apoptosis in other types of cancer cells. Three MB cell lines (DAOY, D283 and D341) were subjected to ATRA treatment. The proportions of cells in the G0/G1 phase of cell cycle and in apoptosis were evaluated. The results showed that cell growth arrest, rather than apoptosis, was the main mechanism by which RA inhibited cell proliferation in the MB cell line DAOY, but not in the others (D283 and D341). Decreased expression of CyclinD1 and C-myc which regulate the transition of cell cycle was observed in DAOY cells following drug treatment, suggesting that these genes might be involved in ATRA retardation of cell cycle progression. Expression of RARbeta, a mediator of the action of retinoids, was also induced by RA in DAOY cells, implying that RAR-beta might also be involved in the mechanism of RA-induced cell cycle arrest. In conclusion, we have provided evidence for the first time that RA may induce cell cycle arrest in vitro in DAOY MB cells via inhibition of CyclinD1 or C-myc. PMID:17453147

Chang, Qing; Chen, Zhengshan; You, Jiangfeng; McNutt, Michael A; Zhang, Ting; Han, Zhihui; Zhang, Xiaoyan; Gong, Encong; Gu, Jiang



Colony Formation from Protoplasts of Nitrate Reductase-deficient Rice Cell Lines.  


Nitrate reductase-deficient cell lines were selected when rice calli grown from microspores by anther culture were inoculated on chlorate medium. 67 calli resistant to chlorate were then transferred to NO(3) medium on which nitrate reductase-deficient cells cannot grow. Calli of two cell lines subsequently did not grow on this medium. These two cell lines, 120-2 and 144, contained no nitrate reductase activity. Xanthine dehydrogenase, but not cytochrome c reductase activity was present in these two lines. Both cell lines consisted of small cell aggregates in young stage. Viable protoplasts were easily isolated from these cell lines. Colonies formed on modified AA medium (Müller and Grafe 1978) containing 2 mg · l(-1) 2,4-D, glucose, and sucrose. These cell lines could be useful for gene manipulation in rice. PMID:23195716

Wakasa, K; Kobayashi, M; Kamada, H



Sedimentation field flow fractionation purification of immature neural cells from a human tumor neuroblastoma cell line.  


The use of stem cells for therapeutic applications is now an important objective for the future. Stem cell preparation is difficult and time-consuming depending on the origin of cells. Sedimentation field flow fractionation (SdFFF) is an effective tool for cell separation, respecting integrity and viability. We used the human neuroblastic SH-SY5Y clone of the SK-N-SH cell line as a source of immature neural cells. Our results demonstrated that by using SdFFF cell sorter under strictly defined conditions, and immunological cell characterization, we are now able to provide, in less than 15 min, a sterile, viable, usable and purified immature neural cell fraction without inducting cell differentiation. PMID:12798175

Lautrette, C; Cardot, P J P; Vermot-Desroches, C; Wijdenes, J; Jauberteau, M O; Battu, S



Characterization of homozygous deletions in laryngeal squamous cell carcinoma cell lines.  


The majority of classical tumor suppressor genes, such as CDKN2A or RB1, were identified by delineation of biallelic losses called homozygous deletions. To systematically identify homozygous deletions in laryngeal squamous cell carcinoma and to unravel novel putative tumor suppressor genes we screened three laryngeal squamous cell carcinoma cell lines (LSCC) using array comparative genomic hybridization (array-CGH). Out of 31 candidate regions for homozygous deletions identified by array-CGH, 5 were verified further by PCR. Among others, these homozygous deletions affected the tumor suppressor gene CDKN2A and the apoptosis-inducing STK17A gene. To assess the frequency of the identified deletions we investigated the affected sites in 9 additional LSCC cell lines. In 5 of the 9 cell lines the CDKN2A gene was homozygously lost. Thus, CDKN2A was homozygously deleted in 7 of the 12 cell lines. No other recurrent homozygous deletions were found. Homozygous deletions was a frequent mechanism of CDKN2A inactivation. Moreover, we identified several other genes, including the putative tumor suppressor gene STK17A, which may be inactivated by homozygous deletions and thus are potentially implicated in laryngeal squamous cell carcinoma development. PMID:18558287

Giefing, Maciej; Martin-Subero, Jose Ignacio; Kiwerska, Katarzyna; Jarmuz, Ma?gorzata; Grenman, Reidar; Siebert, Reiner; Szyfter, Krzysztof



Oxaliplatin, tetraplatin, cisplatin, and carboplatin: Spectrum of activity in drug-resistant cell lines and in the cell lines of the national cancer institute's anticancer drug screen panel  

Microsoft Academic Search

The present study was designed to explore the activity of platinum compounds in cisplatinresistant cell lines, the unselected cell lines of the National Cancer Institute's Anticancer Drug Screen, and the potential for use in combination. The activities of four platinum compounds in cisplatin-resistant KB and A2780 cells were investigated. The cells were highly resistant to cisplatin and cross-resistant to carboplatin,

Olivier Rixe; Waldo Ortuzar; Manuel Alvarez; Ricardo Parker; Eddie Reed; Ken Paull; Tito Fojo



Establishment of a clonal cell line that differentiates into adipose cells in vitro  

Microsoft Academic Search

Summary  From the fibroblastic cells of murine mammary tumor tissue we isolated a clonal cell line that could be induced to differentiate\\u000a into fat-accumulating cells in vitro. Differentiation began after the cultures had reached confluence and was accompanied\\u000a by (a) an increase in the incorporation of sodium acetate into the triglyceride (TG) fraction of cellular lipids, (b) a more\\u000a than 50-fold

Akiyoshi Hiragun; Mayumi Sato; Hiromi Mitsui



Mink Lung Epithelial Cells: Unique Cell Line That Supports Influenza A and B Virus Replication  

PubMed Central

We have demonstrated for the first time that a mink lung epithelial cell line (Mv1Lu) supports the replication of influenza A and B viruses, including the recently isolated H5N1 avian and human Hong Kong strains, to titers comparable to those in MDCK cells. These results suggest that Mv1Lu cells might serve as an alternative system for the isolation and cultivation of influenza A and B viruses and may be useful for vaccine development.

Schultz-Cherry, Stacey; Dybdahl-Sissoko, Naomi; McGregor, Martha; Hinshaw, Virginia S.



Atrazine Exposure Decreases Cell Proliferation in Chinese Hamster Ovary (CHO-K1) Cell Line  

Microsoft Academic Search

The aim of this study was to determine the toxic effect of atrazine at the ovarian cellular level. Chinese Hamster Ovary (CHO-K1) cell line was used to evaluate the degree of in vitro atrazine cytotoxicity and the morphological changes were followed\\u000a during the cell death. Application of four bioassays confirmed that atrazine decreases ovarian cell proliferation and IC50 were determined with

I. Kmeti?; V. Gaurina Sr?ek; I. Slivac; B. Šimi?; Z. Kniewald; J. Kniewald



Susceptibility of Multidrug-resistant Human Leukemia Cell Lines to Human Interleukin 2-activated Killer Cells  

Microsoft Academic Search

Considering the possibility to overcome drug resistance by other treatment strategies than chemotherapy we investigated the susceptibility of three independently selected multidrug-resistant sublines of the T- lymphoblastoid leukemic cell line CCRF-CEM to lymphokine-activated killer (LAK) cells. We found that two of the multidrug-resistant sublines were significantly less susceptible targets to LAK cells. A third one, however, was as susceptible as

Astrid Kimmig; Volker Gekeler; Manfred Neumann; Gerd Frese; Rupert Handgretinger; Gabriella Kardos; Heyke Diddens; Dietrich Niethammer



Characterization of homozygous deletions in laryngeal squamous cell carcinoma cell lines  

Microsoft Academic Search

The majority of classical tumor suppressor genes, such as CDKN2A or RB1, were identified by delineation of biallelic losses called homozygous deletions. To systematically identify homozygous deletions in laryngeal squamous cell carcinoma and to unravel novel putative tumor suppressor genes we screened three laryngeal squamous cell carcinoma cell lines (LSCC) using array comparative genomic hybridization (array-CGH). Out of 31 candidate

Maciej Giefing; Jose Ignacio Martin-Subero; Katarzyna Kiwerska; Ma?gorzata Jarmu?; Reidar Grenman; Reiner Siebert; Krzysztof Szyfter



Multidrug Resistance in a Human Small Cell Lung Cancer Cell Line Selected in Adriamycin1  

Microsoft Academic Search

A multidrug resistant variant (H69AR) of the human small cell lung cancer cell line NCI-H69 was obtained by culturing these cells in grad ually increasing doses of Adriamycin up to 0.8 *tMafter a total of 14 months. H69AR expresses the multidrug resistant phenotype because it is cross-resistant to anthracycline analogues including daunomycin, epi- rubicin, menogaril, and mitoxantrone as well as

L. Mirski; James H. Gerlach; Susan P. C. Cole



Curcumin downregulates cell survival mechanisms in human prostate cancer cell lines  

Microsoft Academic Search

While the role of nuclear transcription factor activator protein-1 (AP-1) in cell proliferation, and of nuclear factor-?B (NF-?B) in the suppression of apoptosis are known, their role in survival of prostate cancer cells is not well understood. We investigated the role of NF-?B and AP-1 in the survival of human androgen-independent (DU145) and -dependent (LNCaP) prostate cancer cell lines. Our

Asok Mukhopadhyay; Carlos Bueso-Ramos; Devasis Chatterjee; Panayotis Pantazis; Bharat B Aggarwal



The biological characteristics of glioma stem cells in human glioma cell line SHG44.  


Gliomas are the most common tumors of the central nervous system (CNS) and a frequent cause of death. The treatment of malignant gliomas is often palliative due to their high recurrence rate. A growing body of evidence suggests that glioma may arise from cancer stem cells (CSC) correlated with neural stem cells (NSC), with the capacity for self-renewal and multipotency. CSCs have been isolated from human gliomas and numerous other solid tumors. It is assumed that a number of established malignant cell lines also contain a rare subpopulation of stem cells. This study was designed to investigate the proportion of CSCs in the human glioma cell line SHG44 and to study the limitations of CD133 immunophenotyping in glioma stem cell research. SHG44 cells were cultured in both serum-containing and serum-free medium. The similar shape in growth curves (in the exponential growth phase) revealed that most cells participated in the population amplification. Time gradient BrdU labeling and monoclonal assay revealed that almost every single cell participated in the division growth (98.82%) and possessed the ability to form clones (96.19%). No significant difference was found in the proportions of CD133+ cells in the serum-containing and serum-free groups (38.25%/37.9