NASA Bioreactor

NASA Technical Reports Server (NTRS)

1998-01-01

Biotechnology Specimen Temperature Controller (BSTC) will cultivate cells until their turn in the bioreactor; it can also be used in culturing experiments that do not require the bioreactor. The BSTC comprises four incubation/refrigeration chambers individually set at 4 to 50 degreesC (near-freezing to above body temperature). Each chamber holds three rugged tissue chamber modules (12 total), clear Teflon bags holding 30 ml of growth media, all positioned by a metal frame. Every 7 to 21 days (depending on growth rates), an astronaut uses a shrouded syringe and the bags' needleless injection ports to transfer a few cells to a fresh media bag, and to introduce a fixative so that the cells may be studied after flight. The design also lets the crew sample the media to measure glucose, gas, and pH levels, and to inspect cells with a microscope. The controller is monitored by the flight crew through a 23-cm (9-inch) color computer display on the face of the BSTC. This view shows the BTSC with the front panel open. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Prostate tumor grown in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2001-01-01

This prostate cancer construct was grown during NASA-sponsored bioreactor studies on Earth. Cells are attached to a biodegradable plastic lattice that gives them a head start in growth. Prostate tumor cells are to be grown in a NASA-sponsored Bioreactor experiment aboard the STS-107 Research-1 mission in 2002. Dr. Leland Chung of the University of Virginia is the principal investigator. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: NASA and the University of Virginia.

Cells growing in NASA Bioreactor

NASA Technical Reports Server (NTRS)

1998-01-01

For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

NASA Classroom Bioreactor

NASA Technical Reports Server (NTRS)

Scully, Robert

2004-01-01

Exploration of space provides a compelling need for cell-based research into the basic mechanisms that underlie the profound changes that occur in terrestrial life that is transitioned to low gravity environments. Toward that end, NASA developed a rotating bioreactor in which cells are cultured while continuously suspended in a cylinder in which the culture medium rotates with the cylinder. The randomization of the gravity vector accomplished by the continuous rotation, in a low shear environment, provides an analog of microgravity. Because cultures grown in bioreactors develop structures and functions that are much closer to those exhibited by native tissue than can be achieved with traditional culture methods, bioreactors have contributed substantially to advancing research in the fields of cancer, diabetes, infectious disease modeling for vaccine production, drug efficacy, and tissue engineering. NASA has developed a Classroom Bioreactor (CB) that is built from parts that are easily obtained and assembled, user-friendly and versatile. It can be easily used in simple school settings to examine the effect cultures of seeds or cells. An educational brief provides assembly instructions and lesson plans that describes activities in science, math and technology that explore free fall, microgravity, orbits, bioreactors, structure-function relationships and the scientific method.

Heart tissue grown in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2001-01-01

Lisa Freed and Gordana Vunjak-Novakovic, both of the Massachusetts Institute of Technology (MIT), have taken the first steps toward engineering heart muscle tissue that could one day be used to patch damaged human hearts. Cells isolated from very young animals are attached to a three-dimensional polymer scaffold, then placed in a NASA bioreactor. The cells do not divide, but after about a week start to cornect to form a functional piece of tissue. Functionally connected heart cells that are capable of transmitting electrical signals are the goal for Freed and Vunjak-Novakovic. Electrophysiological recordings of engineered tissue show spontaneous contractions at a rate of 70 beats per minute (a), and paced contractions at rates of 80, 150, and 200 beats per minute respectively (b, c, and d). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: NASA and MIT.

Heart tissue grown in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2001-01-01

Lisa Freed and Gordana Vunjak-Novakovic, both of the Massachusetts Institute of Technology (MIT), have taken the first steps toward engineering heart muscle tissue that could one day be used to patch damaged human hearts. Cells isolated from very young animals are attached to a three-dimensional polymer scaffold, then placed in a NASA bioreactor. The cells do not divide, but after about a week start to cornect to form a functional piece of tissue. Here, a transmission electron micrograph of engineered tissue shows a number of important landmarks present in functional heart tissue: (A) well-organized myofilaments (Mfl), z-lines (Z), and abundant glycogen granules (Gly); and (D) intercalcated disc (ID) and desmosomes (DES). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: MIT

Colon tumor cells grown in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2001-01-01

These photos compare the results of colon carcinoma cells grown in a NASA Bioreactor flown on the STS-70 Space Shuttle in 1995 flight and ground control experiments. The cells grown in microgravity (left) have aggregated to form masses that are larger and more similar to tissue found in the body than the cells cultured on the ground (right). The principal investigator is Milburn Jessup of the University of Texas M. D. Anderson Cancer Center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Credit: NASA and University of Texas M. D. Anderson Cancer Center.

Tissue grown in space in NASA Bioreactor

NASA Technical Reports Server (NTRS)

1998-01-01

For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, such as the culture section shown here, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. The two white circles within the tumor are part of a plastic lattice that helped the cells associate. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Tissue grown in space in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2001-01-01

Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. Final samples from Mir and Earth appeared histologically cartilaginous throughout their entire cross sections (5-8 mm thick), with the exception of fibrous outer capsules. Constructs grown on Earth (A) appeared to have a more organized extracellular matrix with more uniform collagen orientation as compared with constructs grown on Mir (B), but the average collagen fiber diameter was similar in the two groups (22 +- 2 nm) and comparable to that previously reported for developing articular cartilage. Randomly oriented collagen in Mir samples would be consistent with previous reports that microgravity disrupts fibrillogenesis. These are transmission electron micrographs of constructs from Mir (A) and Earth (B) groups at magnifications of x3,500 and x120,000 (Inset). The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Credit: Proceedings of the National Academy of Sciences.

NASA Bioreactors Advance Disease Treatments

NASA Technical Reports Server (NTRS)

2009-01-01

the body. Experiments conducted by Johnson scientist Dr. Thomas Goodwin proved that the NASA bioreactor could successfully cultivate cells using simulated microgravity, resulting in three-dimensional tissues that more closely approximate those in the body. Further experiments conducted on space shuttle missions and by Wolf as an astronaut on the Mir space station demonstrated that the bioreactor s effects were even further expanded in space, resulting in remarkable levels of tissue formation. While the bioreactor may one day culture red blood cells for injured astronauts or single-celled organisms like algae as food or oxygen producers for a Mars colony, the technology s cell growth capability offers significant opportunities for terrestrial medical research right now. A small Texas company is taking advantage of the NASA technology to advance promising treatment applications for diseases both common and obscure.

Tissue grown in space in NASA Bioreactor

NASA Technical Reports Server (NTRS)

1998-01-01

Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens of cartilage tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. Constructs grown on Mir (A) tended to become more spherical, whereas those grown on Earth (B) maintained their initial disc shape. These findings might be related to differences in cultivation conditions, i.e., videotapes showed that constructs floated freely in microgravity but settled and collided with the rotating vessel wall at 1g (Earth's gravity). In particular, on Mir the constructs were exposed to uniform shear and mass transfer at all surfaces such that the tissue grew equally in all directions, whereas on Earth the settling of discoid constructs tended to align their flat circular areas perpendicular to the direction of motion, increasing shear and mass transfer circumferentially such that the tissue grew preferentially in the radial direction. A and B are full cross sections of constructs from Mir and Earth groups shown at 10-power. C and D are representative areas at the construct surfaces enlarged to 200-power. They are stained red with safranin-O. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). Photo credit: Proceedings of the National Academy of Sciences.

Bioreactor principles

NASA Technical Reports Server (NTRS)

2001-01-01

Cells cultured on Earth (left) typically settle quickly on the bottom of culture vessels due to gravity. In microgravity (right), cells remain suspended and aggregate to form three-dimensional tissue. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Bioreactor

NASA Technical Reports Server (NTRS)

1996-01-01

The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues currently being cultured in rotating bioreactors by investigators

NASA's Bioreactor: Growing Cells in a Simulated Microgravity Environment

NASA Technical Reports Server (NTRS)

Richardson, Denise

2003-01-01

National Science Education Standards (NSES), Science for All Americans, the Secretary's Commission on Achieving Necessary Skills (SCANS) as well as the National Aeronautics and Space Administration (NASA) are all making an effort to promote scientific literacy in America. Unfortunately, major evaluation programs such as the National Assessment of Educational Progress (NAEP) and the Third International Mathematics and Science Study (TIMSS) have provided information that suggested our students are not able to compete with peers from comparable countries. Although results indicated that American students are recalling memorized, factual knowledge well enough, the real problem is the ability to apply what they know. Concerned with these reports, the National Science Teacher's Association (NSTA) has developed a mission to support innovation and high quality in science teaching and learning for every student. NSTA recommends less emphasis on factual knowledge (memorization) and information and more understanding of the concepts. Science process skills are considered imperative to prepare America's students for the 21st century. The National Aeronautics and Space Administration (NASA) supports this mission and adds that NASA strives to help prepare and encourage the next generation of researchers and explorers. One method that NASA supports educators and its mission is to publish educational briefs. NASA describes a brief as a publication that ranges from one-to-thirty pages. The focus is on mission discoveries and results. The brief provides curriculum to educators that supports their objectives and NASA's interest. Educational Briefs are specific to the grade level and course so that educators may have choices that fit their methods and students level. Sometimes, the brief includes lessons and activities teachers may use. For example, NASA's Microgravity Division has designed a student bioreactor. Consequently, an Educational Brief is being written that focuses on how

A Good Neighborhood for Cells: Bioreactor Demonstration System (BDS-05)

NASA Technical Reports Server (NTRS)

Chung, Leland W. K.; Goodwin, Thomas J. (Technical Monitor)

2002-01-01

Good neighborhoods help you grow. As with a city, the lives of a cell are governed by its neighborhood connections Connections that do not work are implicated in a range of diseases. One of those connections - between prostate cancer and bone cells - will be studied on STS-107 using the Bioreactor Demonstration System (BDS-05). To improve the prospects for finding novel therapies, and to identify biomarkers that predict disease progression, scientists need tissue models that behave the same as metastatic or spreading cancer. This is one of several NASA-sponsored lines of cell science research that use the microgravity environment of orbit in an attempt to grow lifelike tissue models for health research. As cells replicate, they "self associate" to form a complex matrix of collagens, proteins, fibers, and other structures. This highly evolved microenvironment tells each cell who is next door, how it should grow arid into what shapes, and how to respond to bacteria, wounds, and other stimuli. Studying these mechanisms outside the body is difficult because cells do not easily self-associate outside a natural environment. Most cell cultures produce thin, flat specimens that offer limited insight into how cells work together. Ironically, growing cell cultures in the microgravity of space produces cell assemblies that more closely resemble what is found in bodies on Earth. NASA's Bioreactor comprises a miniature life support system and a rotating vessel containing cell specimens in a nutrient medium. Orbital BDS experiments that cultured colon and prostate cancers have been highly promising.

Rotating Bioreactor

NASA Technical Reports Server (NTRS)

1988-01-01

The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues currently being cultured in rotating bioreactors by investigators.

Bioreactor rotating wall vessel

NASA Technical Reports Server (NTRS)

2001-01-01

The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

Video of Tissue Grown in Space in NASA Bioreactor

NASA Technical Reports Server (NTRS)

2003-01-01

Principal investigator Leland Chung grew prostate cancer and bone stromal cells aboard the Space Shuttle Columbia during the STS-107 mission. Although the experiment samples were lost along with the ill-fated spacecraft and crew, he did obtain downlinked video of the experiment that indicates the enormous potential of growing tissues in microgravity. Cells grown aboard Columbia had grown far larger tissue aggregates at day 5 than did the cells grown in a NASA bioreactor on the ground.

NASA-approved rotary bioreactor enhances proliferation of human epidermal stem cells and supports formation of 3D epidermis-like structure.

PubMed

Lei, Xiao-hua; Ning, Li-na; Cao, Yu-jing; Liu, Shuang; Zhang, Shou-bing; Qiu, Zhi-fang; Hu, Hui-min; Zhang, Hui-shan; Liu, Shu; Duan, En-kui

2011-01-01

The skin is susceptible to different injuries and diseases. One major obstacle in skin tissue engineering is how to develop functional three-dimensional (3D) substitute for damaged skin. Previous studies have proved a 3D dynamic simulated microgravity (SMG) culture system as a "stimulatory" environment for the proliferation and differentiation of stem cells. Here, we employed the NASA-approved rotary bioreactor to investigate the proliferation and differentiation of human epidermal stem cells (hEpSCs). hEpSCs were isolated from children foreskins and enriched by collecting epidermal stem cell colonies. Cytodex-3 micro-carriers and hEpSCs were co-cultured in the rotary bioreactor and 6-well dish for 15 days. The result showed that hEpSCs cultured in rotary bioreactor exhibited enhanced proliferation and viability surpassing those cultured in static conditions. Additionally, immunostaining analysis confirmed higher percentage of ki67 positive cells in rotary bioreactor compared with the static culture. In contrast, comparing with static culture, cells in the rotary bioreactor displayed a low expression of involucrin at day 10. Histological analysis revealed that cells cultured in rotary bioreactor aggregated on the micro-carriers and formed multilayer 3D epidermis structures. In conclusion, our research suggests that NASA-approved rotary bioreactor can support the proliferation of hEpSCs and provide a strategy to form multilayer epidermis structure.

Osteocytes Mechanosensing in NASA Rotating Wall Bioreactor

NASA Technical Reports Server (NTRS)

Spatz, Jordan; Sibonga, Jean; Wu, Honglu; Barry, Kevin; Bouxsein, Mary; Pajevic, Paola Divieti

2010-01-01

Osteocyte cells are the most abundant (90%) yet least understood bone cell type in the human body. Osteocytes are theorized to be the mechanosensors and transducers of mechanical load for bones, yet the biological mechanism of this action remains elusive. However, recent discoveries in osteocyte cell biology have shed light on their importance as key mechanosensing cells regulating bone remodeling and phosphate homeostasis. The aim of this project was to characterize gene expression patterns and protein levels following exposure of MLO-Y4, a very well characterized murine osteocyte-like cell line, to simulated microgravity using the NASA Rotating Wall Vessel (RWV) Bioreactor. To determine mechanistic pathways of the osteocyte's gravity sensing ability, we evaluated in vitro gene and protein expression of osteocytes exposed to simulated microgravity. Improved understanding of the fundamental mechanisms of mechano transduction at the osteocyte cellular level may lead to revolutionary treatment otions to mitigate the effects of bone loss encountered by astronauts on long duration space missions and provide tailored treatment options for maintaining bone strength of immobilized/partially paralyzed patients here on Earth.

Reduced-Gravity Experiments Conducted to Help Bioreactor Development

NASA Technical Reports Server (NTRS)

Niederhaus, Charles E.; Nahra, Henry K.; Kizito, John P.

2004-01-01

The NASA Glenn Research Center and the NASA Johnson Space Center are collaborating on fluid dynamic investigations for a future cell science bioreactor to fly on the International Space Station (ISS). Project Manager Steven Gonda from the Cellular Biotechnology Program at Johnson is leading the development of the Hydrodynamic Focusing Bioreactor--Space (HFB-S) for use on the ISS to study tissue growth in microgravity. Glenn is providing microgravity fluid physics expertise to help with the design and evaluation of the HFB-S. These bioreactors are used for three-dimensional tissue culture, which cannot be done in ground-based labs in normal gravity. The bioreactors provide a continual supply of oxygen for cell growth, as well as periodic replacement of cell culture media with nutrients. The bioreactor must provide a uniform distribution of oxygen and nutrients while minimizing the shear stresses on the tissue culture.

Long term organ culture of human prostate tissue in a NASA-designed rotating wall bioreactor

NASA Technical Reports Server (NTRS)

Margolis, L.; Hatfill, S.; Chuaqui, R.; Vocke, C.; Emmert-Buck, M.; Linehan, W. M.; Duray, P. H.

1999-01-01

PURPOSE: To maintain ex vivo integral prostatic tissue including intact stromal and ductal elements using the NASA-designed Rotating Wall Vessel (RWV) which maintains colocalized cells in an environment that promotes both three-dimensional cellular interactions together with the uniform mass transfer of nutrients and metabolic wastes. MATERIALS AND METHODS: Samples of normal prostate were obtained as a byproduct of transurethral prostatectomy or needle biopsy. Prostatic tissue dissected into small 1 x 1 mm. blocks was cultured in the Rotating Wall Vessel (RWV) Bioreactor for various time periods and analyzed using histological, immunochemical, and total cell RNA assays. RESULTS: We report the long term maintenance of benign explanted human prostate tissue grown in simple culture medium, under the simulated microgravity conditions afforded by the RWV bioreactor. Mesenchymal stromal elements including blood vessels and architecturally preserved tubuloglandular acini were maintained for a minimum of 28 days. Cytokeratins, vimentin and TGF-beta2 receptor and ligand were preserved through the entire culture period as revealed by immunocytochemistry. Prostatic acid phosphatase (PAP) was continuously expressed during the culture period, although somewhat decreased. Prostatic specific antigen (PSA) and its transcript were down regulated over time of culture. Prostatic carcinoma cells from the TSU cell line were able to invade RWV-cultured benign prostate tissue explants. CONCLUSIONS: The RWV bioreactor represents an additional new technology for culturing prostate tissue for further investigations concerning the basic physiology and pathobiology of this clinically important tissue.

Salmonella Typhimurium grown in a rotating wall bioreactor

NASA Technical Reports Server (NTRS)

2003-01-01

Salmonella typhimurium appears green in on human intestinal tissue (stained red) cultured in a NASA rotating wall bioreactor. Dr. Cheryl Nickerson of Tulane University is studying the effects of simulated low-g on a well-known pathogen, Salmonella typhimurium, a bacterium that causes two to four million cases of gastrointestinal illness in the United States each year. While most healthy people recover readily, S. typhimurium can kill people with weakened immune systems. Thus, a simple case of food poisoning could disrupt a space mission. Using the NASA rotating-wall bioreactor, Nickerson cultured S. typhimurium in modeled microgravity. Mice infected with the bacterium died an average of three days faster than the control mice, indicating that S. typhimurium's virulence was enhanced by the bioreactor. Earlier research showed that 3 percent of the genes were altered by exposure to the bioreactor. Nickerson's work earned her a 2001 Presidential Early Career Award for Scientists and Engineers.

Use of NASA Bioreactor in Engineering Tissue for Bone Repair

NASA Technical Reports Server (NTRS)

Duke, Pauline

1998-01-01

This study was proposed in search for a new alternative for bone replacement or repair. Because the systems commonly used in repair of bony defects form bone by going through a cartilaginous phase, implantation of a piece of cartilage could enhance the healing process by having a more advanced starting point. However, cartilage has seldom been used to replace bone due, in part, to the limitations in conventional culture systems that did not allow production of enough tissue for implants. The NASA-developed bioreactors known as STLV (Slow Turning Lateral Vessel) provide homogeneous distribution of cells, nutrients, and waste products, with less damaging turbulence and shear forces than conventional systems. Cultures under these conditions have higher growth rates, viability, and longevity, allowing larger "tissue-like" aggregates to form, thus opening the possibilities of producing enough tissue for implantation, along with the inherent advantages of in vitro manipulations. To assure large numbers of cells and to eliminate the use of timed embryos, we proposed to use an immortalized mouse limb bud cell line as the source of cells.

Bioreactor Landfill Research and Demonstration Project Northern Oaks Landfill, Harrison, MI

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhao, Xiando; Voice, Thomas; and Hashsham, Syed A.

2006-08-29

A bioreactor landfill cell with 1.2-acre footprint was constructed, filled, operated, and monitored at Northern Oaks Recycling and Disposal Facility (NORDF) at Harrison, MI. With a filled volume of 74,239 cubic yards, the cell contained approximately 35,317 tons of municipal solid waste (MSW) and 20,777 tons of cover soil. It was laid on the slope of an existing cell but separated by a geosynthetic membrane liner. After the cell reached a design height of 60 feet, it was covered with a geosynthetic membrane cap. A three-dimensional monitoring system to collect data at 48 different locations was designed and installed duringmore » the construction phase of the bioreactor cell. Each location had a cluster of monitoring devices consisting of a probe to monitor moisture and temperature, a leachate collection basin, and a gas sampling port. An increase in moisture content of the MSW in the bioreactor cell was achieved by pumping leachate collected on-site from various other cells, as well as recirculation of leachate from the bioreactor landfill cell itself. Three types of leachate injection systems were evaluated in this bioreactor cell for their efficacy to distribute pumped leachate uniformly: a leachate injection pipe buried in a 6-ft wide horizontal stone mound, a 15-ft wide geocomposite drainage layer, and a 60-ft wide geocomposite drainage layer. All leachate injection systems were installed on top of the compacted waste surface. The distribution of water and resulting MSW moisture content throughout the bioreactor cell was found to be similar for the three designs. Water coming into and leaving the cell (leachate pumped in, precipitation, snow, evaporation, and collected leachate) was monitored in order to carry out a water balance. Using a leachate injection rate of 26 – 30 gal/yard3, the average moisture content increased from 25% to 35% (wet based) over the period of this study. One of the key aspects of this bioreactor landfill study was to evaluate

NASA Technology Demonstrations Missions Program Overview

NASA Technical Reports Server (NTRS)

Turner, Susan

2011-01-01

The National Aeronautics and Space Administration (NASA) Fiscal Year 2010 (FY10) budget introduced a new strategic plan that placed renewed emphasis on advanced missions beyond Earth orbit. This supports NASA s 2011 strategic goal to create innovative new space technologies for our exploration, science, and economic future. As a result of this focus on undertaking many and more complex missions, NASA placed its attention on a greater investment in technology development, and this shift resulted in the establishment of the Technology Demonstrations Missions (TDM) Program. The TDM Program, within the newly formed NASA Office of the Chief Technologist, supports NASA s grand challenges by providing a steady cadence of advanced space technology demonstrations (Figure 1), allowing the infusion of flexible path capabilities for future exploration. The TDM Program's goal is to mature crosscutting capabilities to flight readiness in support of multiple future space missions, including flight test projects where demonstration is needed before the capability can transition to direct mission The TDM Program has several unique criteria that set it apart from other NASA program offices. For instance, the TDM Office matures a small number of technologies that are of benefit to multiple customers to flight technology readiness level (TRL) 6 through relevant environment testing on a 3-year development schedule. These technologies must be crosscutting, which is defined as technology with potential to benefit multiple mission directorates, other government agencies, or the aerospace industry, and they must capture significant public interest and awareness. These projects will rely heavily on industry partner collaboration, and funding is capped for all elements of the flight test demonstration including planning, hardware development, software development, launch costs, ground operations, and post-test assessments. In order to inspire collaboration across government and industry

Some process control/design considerations in the development of a microgravity mammalian cell bioreactor

NASA Technical Reports Server (NTRS)

Goochee, Charles F.

1987-01-01

The purpose is to review some of the physical/metabolic factors which must be considered in the development of an operating strategy for a mammalian cell bioreactor. Emphasis is placed on the dissolved oxygen and carbon dioxide requirements of growing mammalian epithelial cells. Literature reviews concerning oxygen and carbon dioxide requirements are discussed. A preliminary, dynamic model which encompasses the current features of the NASA bioreactor is presented. The implications of the literature survey and modeling effort on the design and operation of the NASA bioreactor are discussed.

Enhanced biodegradation of methylhydrazine and hydrazine contaminated NASA wastewater in fixed-film bioreactor.

PubMed

Nwankwoala, A U; Egiebor, N O; Nyavor, K

2001-01-01

The aerobic biodegradation of National Aeronautics and Space Administration (NASA) wastewater that contains mixtures of highly concentrated methylhydrazine/hydrazine, citric acid and their reaction product was studied on a laboratory-scale fixed film trickle-bed reactor. The degrading organisms, Achromobacter sp., Rhodococcus B30 and Rhodococcus J10, were immobilized on coarse sand grains used as support-media in the columns. Under continuous flow operation, Rhodococcus sp. degraded the methylhydrazine content of the wastewater from a concentration of 10 to 2.5 mg/mL within 12 days and the hydrazine from approximately 0.8 to 0.1 mg/mL in 7 days. The Achromobacter sp. was equally efficient in degrading the organics present in the wastewater, reducing the concentration of the methylhydrazine from 10 to approximately 5 mg/mL within 12 days and that of the hydrazine from approximately 0.8 to 0.2 mg/mL in 7 days. The pseudo first-order rate constants of 0.137 day(-1) and 0.232 day(-1) were obtained for the removal of methylhydrazine and hydrazine, respectively, in wastewater in the reactor column. In the batch cultures, rate constants for the degradation were 0.046 and 0.079 day(-1) for methylhydrazine and hydrazine respectively. These results demonstrate that the continuous flow bioreactor afford greater degradation efficiencies than those obtained when the wastewater was incubated with the microbes in growth-limited batch experiments. They also show that wastewater containing hydrazine is more amenable to microbial degradation than one that is predominant in methylhydrazine, in spite of the longer lag period observed for hydrazine containing wastewater. The influence of substrate concentration and recycle rate on the degradation efficiency is reported. The major advantages of the trickle-bed reactor over the batch system include very high substrate volumetric rate of turnover, higher rates of degradation and tolerance of the 100% concentrated NASA wastewater. The

Erythroid cell growth and differentiation in vitro in the simulated microgravity environment of the NASA rotating wall vessel bioreactor

NASA Technical Reports Server (NTRS)

Sytkowski, A. J.; Davis, K. L.

2001-01-01

Prolonged exposure of humans and experimental animals to the altered gravitational conditions of space flight has adverse effects on the lymphoid and erythroid hematopoietic systems. Although some information is available regarding the cellular and molecular changes in lymphocytes exposed to microgravity, little is known about the erythroid cellular changes that may underlie the reduction in erythropoiesis and resultant anemia. We now report a reduction in erythroid growth and a profound inhibition of erythropoietin (Epo)-induced differentiation in a ground-based simulated microgravity model system. Rauscher murine erythroleukemia cells were grown either in tissue culture vessels at 1 x g or in the simulated microgravity environment of the NASA-designed rotating wall vessel (RWV) bioreactor. Logarithmic growth was observed under both conditions; however, the doubling time in simulated microgravity was only one-half of that seen at 1 x g. No difference in apoptosis was detected. Induction with Epo at the initiation of the culture resulted in differentiation of approximately 25% of the cells at 1 x g, consistent with our previous observations. In contrast, induction with Epo at the initiation of simulated microgravity resulted in only one-half of this degree of differentiation. Significantly, the growth of cells in simulated microgravity for 24 h prior to Epo induction inhibited the differentiation almost completely. The results suggest that the NASA RWV bioreactor may serve as a suitable ground-based microgravity simulator to model the cellular and molecular changes in erythroid cells observed in true microgravity.

NASA experiments onboard the controlled impact demonstration

NASA Technical Reports Server (NTRS)

Hayduk, R. J.; Alfaro-Bou, E.; Fasanella, E. L.

1985-01-01

The structural crashworthiness tests conducted by NASA on the December 1, 1984 controlled impact demonstration are discussed. The components and locations of the data acquisition and photographic systems developed by NASA to evaluate impact loads throughout the aircraft structure and the transmission of loads into the dummies are described. The effectiveness of the NASA designed absorbing seats and the vertical, longitudinal, and transverse impact loads are measured. Data that is extremely applicable to crash dynamics structural research was obtained by the data acquisition system and very low load levels were measured for the NASA energy absorbing seats.

Growing Three-Dimensional Corneal Tissue in a Bioreactor

NASA Technical Reports Server (NTRS)

Spaulding, Glen F.; Goodwin, Thomas J.; Aten, Laurie; Prewett, Tacey; Fitzgerald, Wendy S.; OConnor, Kim; Caldwell, Delmar; Francis, Karen M.

2003-01-01

Spheroids of corneal tissue about 5 mm in diameter have been grown in a bioreactor from an in vitro culture of primary rabbit corneal cells to illustrate the production of optic cells from aggregates and tissue. In comparison with corneal tissues previously grown in vitro by other techniques, this tissue approximates intact corneal tissue more closely in both size and structure. This novel three-dimensional tissue can be used to model cell structures and functions in normal and abnormal corneas. Efforts continue to refine the present in vitro method into one for producing human corneal tissue to overcome the chronic shortage of donors for corneal transplants: The method would be used to prepare corneal tissues, either from in vitro cultures of a patient s own cells or from a well-defined culture from another human donor known to be healthy. As explained in several articles in prior issues of NASA Tech Briefs, generally cylindrical horizontal rotating bioreactors have been developed to provide nutrient-solution environments conducive to the 30 NASA Tech Briefs, October 2003 growth of delicate animal cells, with gentle, low-shear flow conditions that keep the cells in suspension without damaging them. The horizontal rotating bioreactor used in this method, denoted by the acronym "HARV," was described in "High-Aspect-Ratio Rotating Cell-Culture Vessel" (MSC-21662), NASA Tech Briefs, Vol. 16, No. 5 (May, 1992), page 150.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Human primary breast tumor cells after 49 days of growth in a NASA Bioreactor. Tumor cells aggregate on microcarrier beads (indicated by arrow). NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Jearne Becker, University of South Florida

NASA Ground-Truthing Capabilities Demonstrated

NASA Technical Reports Server (NTRS)

Lopez, Isaac; Seibert, Marc A.

2004-01-01

NASA Research and Education Network (NREN) ground truthing is a method of verifying the scientific validity of satellite images and clarifying irregularities in the imagery. Ground-truthed imagery can be used to locate geological compositions of interest for a given area. On Mars, astronaut scientists could ground truth satellite imagery from the planet surface and then pinpoint optimum areas to explore. These astronauts would be able to ground truth imagery, get results back, and use the results during extravehicular activity without returning to Earth to process the data from the mission. NASA's first ground-truthing experiment, performed on June 25 in the Utah desert, demonstrated the ability to extend powerful computing resources to remote locations. Designed by Dr. Richard Beck of the Department of Geography at the University of Cincinnati, who is serving as the lead field scientist, and assisted by Dr. Robert Vincent of Bowling Green State University, the demonstration also involved researchers from the NASA Glenn Research Center and the NASA Ames Research Center, who worked with the university field scientists to design, perform, and analyze results of the experiment. As shown real-time Hyperion satellite imagery (data) is sent to a mass storage facility, while scientists at a remote (Utah) site upload ground spectra (data) to a second mass storage facility. The grid pulls data from both mass storage facilities and performs up to 64 simultaneous band ratio conversions on the data. Moments later, the results from the grid are accessed by local scientists and sent directly to the remote science team. The results are used by the remote science team to locate and explore new critical compositions of interest. The process can be repeated as required to continue to validate the data set or to converge on alternate geophysical areas of interest.

Human cell culture in a space bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R.

1988-01-01

Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

NASA Lidar system support and MOPA technology demonstration

NASA Technical Reports Server (NTRS)

Laughman, L. M.; Capuano, B.; Wayne, R. J.

1986-01-01

A series of lidar design and technology demonstration tasks in support of a CO2 lidar program is discussed. The first of these tasks is discussed in Section VI of this report under the heading of NASA Optical Lidar Design and it consists of detailed recommendations for the layout of a CO2 Doppler lidar incorporating then existing NASA optical components and mounts. The second phase of this work consisted of the design, development, and delivery to NASA of a novel acousto-optic laser frequency stabilization system for use with the existing NASA ring laser transmitter. The second major task in this program encompasses the design and experimental demonstration of a master oscillator-power amplifier (MOPA) laser transmitter utilizing a commercially available laser as the amplifier. The MOPA design including the low chirp master oscillator is discussed in detail. Experimental results are given for one, two and three pass amplification. The report includes operating procedures for the MOPA system.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

High magnification view of human primary breast tumor cells after 56 days of culture in a NASA Bioreactor. The arrow points to bead surface indicating breast cancer cells (as noted by the staining of tumor cell intermediate filaments). NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Jearne Becker, University of South Florida

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

High magnification of view of tumor cells aggregate on microcarrier beads, illustrting breast cells with intercellular boundaires on bead surface and aggregates of cells achieving 3-deminstional growth outward from bead after 56 days of culture in a NASA Bioreactor. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Jearne Becker, University of South Florida.

EVALUATION PLAN FOR TWO LARGE-SCALE LANDFILL BIOREACTOR TECHNOLOGIES

EPA Science Inventory

Abstract - Waste Management, Inc., is operating two long-term bioreactor studies at the Outer Loop Landfill in Louisville, KY, including facultative landfill bioreactor and staged aerobic-anaerobic landfill bioreactor demonstrations. A Quality Assurance Project Plan (QAPP) was p...

NASA's Bioreactor: Growing Cells in a Microgravity Environment. Educational Brief.

ERIC Educational Resources Information Center

National Aeronautics and Space Administration, Washington, DC.

This brief discusses growing cells in a microgravity environment for grades 9-12. Students are provided with plans for building a classroom bioreactor that can then be used with the included activity on seed growth in a microgravity environment. Additional experimental ideas are also suggested along with a history and background on microgravity…

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Time-lapse exposure depicts Bioreactor rotation. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunourous tissues.

BIOREACTOR DESIGN - OUTER LOOP LANDFILL, LOUISVILLE, KY

EPA Science Inventory

Bioreactor field demonstration projects are underway at the Outer Loop Landfill in Louisville, KY, USA. The research effort is a cooperative research effort between US EPA and Waste Management Inc. Two primary kinds of municipal waste bioreactors are under study at this site. ...

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Human primary breast tumor cells after 56 days of culture in a NASA Bioreactor. A cross-section of a construct, grown from surgical specimens of brease cancer, stained for microscopic examination, reveals areas of tumor cells dispersed throughout the non-epithelial cell background. The arrow denotes the foci of breast cancer cells. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Jearne Becker, University of South Florida

Suspension cell culture in microgravity and development of a space bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R.

1987-01-01

NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Dr. Harry Mahtani analyzes the gas content of nutrient media from Bioreactor used in research on human breast cancer. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunourous tissues.

Human Peripheral Blood Mononuclear Cells Cultured in Normal and Hyperglycemic Media in Simulated Microgravity Using NASA Bioreactors

NASA Technical Reports Server (NTRS)

Lawless, DeSales

2003-01-01

We sought answers to several questions this summer at NASA Johnson Space Center. Initial studies involved the in vitro culture of human peripheral blood mononuclear in cells in different conditioned culture media. Several human cancer clones were similarly studied to determine responses to aberrant glycosylation by the argon laser. The cells were grown at unit gravity in flasks and in simulated microgravity using NASA bioreactors. The cells in each instance were analyzed by flow cytometry. Cell cycle analysis was acquired by staining nuclear DNA with propidium iodide. Responses to the laser stimulation was measured by observing autofluorescence emitted in the green and red spectra after stimulation. Extent of glycosylation correlated with the intensity of the laser stimulated auto-fluorescence. Our particular study was to detect and monitor aberrant glycosylation and its role in etiopathogenesis. Comparisons were made between cells known to be neoplastic and normal cell controls using the same Laser Induced Autofluorescence technique. Studies were begun after extensive literature searches on using the antigen presenting potential of dendritic cells to induce proliferation of antigen specific cytotoxic T-cells. The Sendai virus served as the antigen. Our goal is to generate sufficient numbers of such cells in the simulated microgravity environment for use in autologous transplants of virally infected individuals including those positive for hepatitis and HIV.

CECE: A Deep Throttling Demonstrator Cryogenic Engine for NASA's Lunar Lander

NASA Technical Reports Server (NTRS)

Giuliano, Victor J.; Leonard, Timothy G.; Adamski, Walter M.; Kim, Tony S.

2007-01-01

As one of the first technology development programs awarded under NASA's Vision for Space Exploration, the Pratt & Whitney Rocketdyne (PWR) Deep Throttling, Common Extensible Cryogenic Engine (CECE) program was selected by NASA in November 2004 to begin technology development and demonstration toward a deep throttling, cryogenic Lunar Lander engine for use across multiple human and robotic lunar exploration mission segments with extensibility to Mars. The CECE program leverages the maturity and previous investment of a flight-proven hydrogen/oxygen expander cycle engine, the RL10, to develop and demonstrate an unprecedented combination of reliability, safety, durability, throttlability, and restart capabilities in a high-energy, cryogenic engine. NASA Marshall Space Flight Center and NASA Glenn Research Center personnel were integral design and analysis team members throughout the requirements assessment, propellant studies and the deep throttling demonstrator elements of the program. The testbed selected for the initial deep throttling demonstration phase of this program was a minimally modified RL10 engine, allowing for maximum current production engine commonality and extensibility with minimum program cost. In just nine months from technical program start, CECE Demonstrator No. 1 engine testing in April/May 2006 at PWR's E06 test stand successfully demonstrated in excess of 10:1 throttling of the hydrogen/oxygen expander cycle engine. This test provided an early demonstration of a viable, enabling cryogenic propulsion concept with invaluable system-level technology data acquisition toward design and development risk mitigation for both the subsequent CECE Demonstrator No. 2 program and to the future Lunar Lander Design, Development, Test and Evaluation effort.

NASA Habitat Demonstration Unit (HDU) Deep Space Habitat Analog

NASA Technical Reports Server (NTRS)

Howe, A. Scott; Kennedy, Kriss J.; Gill, Tracy

2013-01-01

The NASA Habitat Demonstration Unit (HDU) vertical cylinder habitat was established as a exploration habitat testbed platform for integration and testing of a variety of technologies and subsystems that will be required in a human-occupied planetary surface outpost or Deep Space Habitat (DSH). The HDU functioned as a medium-fidelity habitat prototype from 2010-2012 and allowed teams from all over NASA to collaborate on field analog missions, mission operations tests, and system integration tests to help shake out equipment and provide feedback for technology development cycles and crew training. This paper documents the final 2012 configuration of the HDU, and discusses some of the testing that took place. Though much of the higher-fidelity functionality has 'graduated' into other NASA programs, as of this writing the HDU, renamed Human Exploration Research Analog (HERA), will continue to be available as a volumetric and operational mockup for NASA Human Research Program (HRP) research from 2013 onward.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Epithelial and fibroblast cell coculture: Long-term growth human mammary epithelial cells (HMEC) admixed in coculture with fibroblast from the same initial breast tissue grown as 3-dimenstional constructions in the presence of attachment beads in the NASA Bioreactor. A: A typical constrct about 2.0 mm in diameter without beads on the surface. The center of these constrcts is hollow, and beads are organized about the irner surface. Although the coculture provides smaller constructs than the monoculture, the metabolic of the organized cells is about the same. B, C, D: Closer views of cells showing that the shape of cells and cell-to-cell interactions apprear different in the coculture than in the monoculture constructs. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Richmond, NASA/Marshall Space Flight Center (MSFC).

Bioreactor Scalability: Laboratory-Scale Bioreactor Design Influences Performance, Ecology, and Community Physiology in Expanded Granular Sludge Bed Bioreactors

PubMed Central

Connelly, Stephanie; Shin, Seung G.; Dillon, Robert J.; Ijaz, Umer Z.; Quince, Christopher; Sloan, William T.; Collins, Gavin

2017-01-01

Studies investigating the feasibility of new, or improved, biotechnologies, such as wastewater treatment digesters, inevitably start with laboratory-scale trials. However, it is rarely determined whether laboratory-scale results reflect full-scale performance or microbial ecology. The Expanded Granular Sludge Bed (EGSB) bioreactor, which is a high-rate anaerobic digester configuration, was used as a model to address that knowledge gap in this study. Two laboratory-scale idealizations of the EGSB—a one-dimensional and a three- dimensional scale-down of a full-scale design—were built and operated in triplicate under near-identical conditions to a full-scale EGSB. The laboratory-scale bioreactors were seeded using biomass obtained from the full-scale bioreactor, and, spent water from the distillation of whisky from maize was applied as substrate at both scales. Over 70 days, bioreactor performance, microbial ecology, and microbial community physiology were monitored at various depths in the sludge-beds using 16S rRNA gene sequencing (V4 region), specific methanogenic activity (SMA) assays, and a range of physical and chemical monitoring methods. SMA assays indicated dominance of the hydrogenotrophic pathway at full-scale whilst a more balanced activity profile developed during the laboratory-scale trials. At each scale, Methanobacterium was the dominant methanogenic genus present. Bioreactor performance overall was better at laboratory-scale than full-scale. We observed that bioreactor design at laboratory-scale significantly influenced spatial distribution of microbial community physiology and taxonomy in the bioreactor sludge-bed, with 1-D bioreactor types promoting stratification of each. In the 1-D laboratory bioreactors, increased abundance of Firmicutes was associated with both granule position in the sludge bed and increased activity against acetate and ethanol as substrates. We further observed that stratification in the sludge-bed in 1-D laboratory

Multifunctional Bioreactor System for Human Intestine Tissues

PubMed Central

2017-01-01

The three-dimensional (3D) cultivation of intestinal cells and tissues in dynamic bioreactor systems to represent in vivo intestinal microenvironments is essential for developing regenerative medicine treatments for intestinal diseases. We have previously developed in vitro human intestinal tissue systems using a 3D porous silk scaffold system with intestinal architectures and topographical features for the adhesion, growth, and differentiation of intestinal cells under static culture conditions. In this study, we designed and fabricated a multifunctional bioreactor system that incorporates pre-epithelialized 3D silk scaffolds in a dynamic culture environment for in vitro engineering of human intestine tissues. The bioreactor system allows for control of oxygen levels in perfusion fluids (aerobic simulated intestinal fluid (SIF), microaerobic SIF, and anaerobic SIF), while ensuring control over the mechanical and chemical microenvironments present in native human intestines. The bioreactor system also enables 3D cell culture with spatial separation and cultivation of cocultured epithelial and stromal cells. Preliminary functional analysis of tissues housed in the bioreactor demonstrated that the 3D tissue constructs survived and maintained typical phenotypes of intestinal epithelium, including epithelial tight junction formation, intestinal biomarker expression, microvilli formation, and mucus secretion. The unique combination of a dynamic bioreactor and 3D intestinal constructs offers utility for engineering human intestinal tissues for the study of intestinal diseases and discovery options for new treatments. PMID:29333491

NASA Solar Array Demonstrates Commercial Potential

NASA Technical Reports Server (NTRS)

Creech, Gray

2006-01-01

A state-of-the-art solar-panel array demonstration site at NASA's Dryden Flight Research Center provides a unique opportunity for studying the latest in high-efficiency solar photovoltaic cells. This five-kilowatt solar-array site (see Figure 1) is a technology-transfer and commercialization success for NASA. Among the solar cells at this site are cells of a type that was developed in Dryden Flight Research Center s Environmental Research Aircraft and Sensor Technology (ERAST) program for use in NASA s Helios solar-powered airplane. This cell type, now denoted as A-300, has since been transferred to SunPower Corporation of Sunnyvale, California, enabling mass production of the cells for the commercial market. High efficiency separates these advanced cells from typical previously commercially available solar cells: Whereas typical previously commercially available cells are 12 to 15 percent efficient at converting sunlight to electricity, these advanced cells exhibit efficiencies approaching 23 percent. The increase in efficiency is due largely to the routing of electrical connections behind the cells (see Figure 2). This approach to increasing efficiency originated as a solution to the problem of maximizing the degree of utilization of the limited space available atop the wing of the Helios airplane. In retrospect, the solar cells in use at this site could be used on Helios, but the best cells otherwise commercially available could not be so used, because of their lower efficiencies. Historically, solar cells have been fabricated by use of methods that are common in the semiconductor industry. One of these methods includes the use of photolithography to define the rear electrical-contact features - diffusions, contact openings, and fingers. SunPower uses these methods to produce the advanced cells. To reduce fabrication costs, SunPower continues to explore new methods to define the rear electrical-contact features. The equipment at the demonstration site includes

Cell Separations in Microgravity and Development of a Space Bioreactor

NASA Technical Reports Server (NTRS)

Morrison, D. R.

1985-01-01

A bioreactor optimized for operations in space is now being developed. The current research is focused on determining the optimum cell-bead ratios, medium content and proper maintenance conditions required to keep living cell specimens alive and healthy for the entire flight. The bioreactor development project has recently added a microprocessor/computer to the JSC prototype for control and data analysis. Appropriate new technology is being combined with the current bioreactor designs and tested to determine what specific features must be included in the fabrication of a bioreactor designed to operate for STS demonstration tests. Considerations include: (1) circulation and resupply of culture media; (2) sensors required to monitor temperature, cell growth, mass transport, and oxygen consumption; and (3) inflight control of shear stress on cells, gas transfer in microgravity, diffusion, and intracellular transport. These data and results from the JSC prototype bioreactor test will be used for the design and construction of a small space bioreactor for the Orbiter middeck.

Bioreactor technology for production of valuable algal products

NASA Astrophysics Data System (ADS)

Liu, Guo-Cai; Cao, Ying

1998-03-01

Bioreactor technology has long been employed for the production of various (mostly cheap) food and pharmaceutical products. More recently, research has been mainly focused on the development of novel bioreactor technology for the production of high—value products. This paper reports the employment of novel bioreactor technology for the production of high-value biomass and metabolites by microalgae. These high-value products include microalgal biomass as health foods, pigments including phycocyanin and carotenoids, and polyunsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid. The processes involved include heterotrophic and mixotrophic cultures using organic substrates as the carbon source. We have demonstrated that these bioreactor cultivation systems are particularly suitable for the production of high-value products from various microalgae. These cultivation systems can be further modified to improve cell densities and productivities by using high cell density techniques such as fed-batch and membrane cell recycle systems. For most of the microalgae investigated, the maximum cell concentrations obtained using these bioreactor systems in our laboratories are much higher than any so far reported in the literature.

Bio-reactor chamber

NASA Technical Reports Server (NTRS)

Chandler, Joseph A. (Inventor)

1989-01-01

A bioreactor for cell culture is disclosed which provides for the introduction of fresh medium without excessive turbulent action. The fresh medium enters the bioreactor through a filter with a backwash action which prevents the cells from settling on the filter. The bioreactor is sealed and depleted medium is forced out of the container as fresh medium is added.

Lithium-Ion Battery Demonstrated for NASA Desert Research and Technology Studies

NASA Technical Reports Server (NTRS)

Bennett, William R.; Baldwin, Richard S.

2008-01-01

Lithium-ion batteries have attractive performance characteristics that are well suited to a number of NASA applications. These rechargeable batteries produce compact, lightweight energy-storage systems with excellent cycle life, high charge/discharge efficiency, and low self-discharge rate. NASA Glenn Research Center's Electrochemistry Branch designed and produced five lithium-ion battery packs configured to power the liquid-air backpack (LAB) on spacesuit simulators. The demonstration batteries incorporated advanced, NASA-developed electrolytes with enhanced low-temperature performance characteristics. The objectives of this effort were to (1) demonstrate practical battery performance under field-test conditions and (2) supply laboratory performance data under controlled laboratory conditions. Advanced electrolyte development is being conducted under the Exploration Technology Development Program by the NASA Jet Propulsion Laboratory. Three field trials were successfully completed at Cinder Lake from September 10 to 12, 2007. Extravehicular activities of up to 1 hr and 50 min were supported, with residual battery capacity sufficient for 30 min of additional run time. Additional laboratory testing of batteries and cells is underway at Glenn s Electrochemical Branch.

Latest Changes to NASA's Laser Communication Relay Demonstration Project

NASA Technical Reports Server (NTRS)

Edwards, Bernard L.; Israel, David J.; Vithlani, Seema K.

2018-01-01

Over the last couple of years, NASA has been making changes to the Laser Communications Relay Demonstration Project (LCRD), a joint project between NASA's Goddard Space Flight Center (GSFC), the Jet Propulsion Laboratory, California Institute of Technology (JPL), and the Massachusetts Institute of Technology Lincoln Laboratory (MIT/LL). The changes made makes LCRD more like a future Earth relay system that has both high speed optical and radio frequency links. This will allow LCRD to demonstrate a more detailed concept of operations for a future operational mission critical Earth relay. LCRD is expected to launch in June 2019 and is expected to be followed a couple of years later with a prototype user terminal on the International Space Station. LCRD's architecture will allow it to serve as a testbed in space and this paper will provide an update of its planned capabilities and experiments.

MONITORING GUIDANCE FOR BIOREACTOR LANDFILLS

EPA Science Inventory

Experimental bioreactor landfill operations at operating Municipal Solid Waste (MSW) landfills can be approved under the research development and demonstration (RD&D) provisions of 30CFR 258.4. To provide a basis for consistent data collection for future decision-making in suppor...

MONITORING APPROACHES FOR BIOREACTOR LANDFILLS

EPA Science Inventory

Experimental bioreactor landfill operations at operating Municipal Solid Waste (MSW) landfills can be approved under the research development and demonstration (RD&D) provisions of 40 CFR 258.4. To provide a basis for consistent data collection for future decision-making in suppo...

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Epithelial cell monoculture: Long-term growth of human mammary epithelial cells (HMEC) grown in monoculture as 3-dimensional constructions in the presence of attachment beads in the NASA Bioreactor. A: A typical construct about 3.5 mm (less than 1/8th inch) in diameter with slightly dehydrted, crinkled beads contained on the surface as well as within the 3-dimensional structure. B: The center of these constructs is hollow. Crinkling of the beads causes a few to fall out, leaving crater-like impressiions in the construct. The central impression shows a small hole that accesses the hollow center of the construct. C: A closeup view of the cells and the hole the central impression. D: Closer views of cells in the construct showing sell-to-cell interactions. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Richmond, NASA/Marshall Space Flight Center (MSFC).

Demonstrating Robotic Autonomy in NASA's Intelligent Systems Project

NASA Technical Reports Server (NTRS)

Morris, Robert; Smith, Ben; Estlin, Tara; Pedersen, Liam

2004-01-01

This paper will provide an overview of NASA's investments in autonomy during the past five years within the Intelligent Systems Project, with particular attention paid to investments that have resulted in mission infusion of autonomy technology, in particular, into the recent Mars Exploration Rover (MER) mission. The content of the paper will be divided into two primary topic areas: a technical overview of the component technologies developed under the program, and a programmatic overview of the history and organization of the NASA IS project itself, with a focus on describing the program elements related to autonomy and intelligent robotics. The paper will also provide an overview of the September 2004 autonomy demonstrations, including a discussion of objectives, organization, and preliminary results (to the extent they are available before the submission deadline).

Visualizing medium and biodistribution in complex cell culture bioreactors using in vivo imaging.

PubMed

Ratcliffe, E; Thomas, R J; Stacey, A J

2014-01-01

There is a dearth of technology and methods to aid process characterization, control and scale-up of complex culture platforms that provide niche micro-environments for some stem cell-based products. We have demonstrated a novel use of 3d in vivo imaging systems to visualize medium flow and cell distribution within a complex culture platform (hollow fiber bioreactor) to aid characterization of potential spatial heterogeneity and identify potential routes of bioreactor failure or sources of variability. This can then aid process characterization and control of such systems with a view to scale-up. Two potential sources of variation were observed with multiple bioreactors repeatedly imaged using two different imaging systems: shortcutting of medium between adjacent inlet and outlet ports with the potential to create medium gradients within the bioreactor, and localization of bioluminescent murine 4T1-luc2 cells upon inoculation with the potential to create variable seeding densities at different points within the cell growth chamber. The ability of the imaging technique to identify these key operational bioreactor characteristics demonstrates an emerging technique in troubleshooting and engineering optimization of bioreactor performance. © 2013 American Institute of Chemical Engineers.

Cultivation of mammalian cells using a single-use pneumatic bioreactor system.

PubMed

Obom, Kristina M; Cummings, Patrick J; Ciafardoni, Janelle A; Hashimura, Yasunori; Giroux, Daniel

2014-10-10

Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized medicine innovations. However, translating these advances into therapeutic applications will require in vitro systems that allow for robust, flexible, and cost effective bioreactor systems. There are several bioreactor systems currently utilized in research and commercial settings; however, many of these systems are not optimal for establishing, expanding, and monitoring the growth of different cell types. The culture parameters most challenging to control in these systems include, minimizing hydrodynamic shear, preventing nutrient gradient formation, establishing uniform culture medium aeration, preventing microbial contamination, and monitoring and adjusting culture conditions in real-time. Using a pneumatic single-use bioreactor system, we demonstrate the assembly and operation of this novel bioreactor for mammalian cells grown on micro-carriers. This bioreactor system eliminates many of the challenges associated with currently available systems by minimizing hydrodynamic shear and nutrient gradient formation, and allowing for uniform culture medium aeration. Moreover, the bioreactor's software allows for remote real-time monitoring and adjusting of the bioreactor run parameters. This bioreactor system also has tremendous potential for scale-up of adherent and suspension mammalian cells for production of a variety therapeutic proteins, monoclonal antibodies, stem cells, biosimilars, and vaccines.

Lessons Learned from Inlet Integration Analysis of NASA's Low Boom Flight Demonstrator

NASA Technical Reports Server (NTRS)

Friedlander, David; Heath, Christopher; Castner, Ray

2017-01-01

In 2016, NASA's Aeronautics Research Mission Directorate announced the New Aviation Horizons Initiative with a goal of designing/building several X-Planes, including a Low Boom Flight Demonstrator (LBFD). That same year, NASA awarded a contract to Lockheed Martin (LM) to advance the LBFD concept through preliminary design. Several configurations of the LBFD aircraft were analyzed by both LM engineers and NASA researchers. This presentation focuses on some of the CFD simulations that were run by NASA Glenn researchers. NASA's FUN3D V13.1 code was used for all adjoint-based grid refinement studies and Spalart-Allmaras turbulence model was used during adaptation. It was found that adjoint-based grid adaptation did not accurately capture inlet performance for high speed top-aft-mounted propulsion.

Use of in-field bioreactors demonstrate groundwater filtration influences planktonic bacterial community assembly, but not biofilm composition

DOE PAGES

Christensen, Geoffrey A.; Moon, Ji Won; Veach, Allison M.; ...

2018-03-20

Using in-field bioreactors, we investigated the influence of exogenous microorganisms in groundwater planktonic and biofilm microbial communities as part of the Integrated Field Research Challenge (IFRC). After an acclimation period with source groundwater, bioreactors received either filtered (0.22 μM filter) or unfiltered well groundwater in triplicate and communities were tracked routinely for 23 days after filtration was initiated. To address geochemical influences, the planktonic phase was assayed periodically for protein, organic acids, physico-/geochemical measurements and bacterial community (via 16S rRNA gene sequencing), while biofilms (i.e. microbial growth on sediment coupons) were targeted for bacterial community composition at the completion ofmore » the experiment (23 d). Based on Bray-Curtis distance, planktonic bacterial community composition varied temporally and between treatments (filtered, unfiltered bioreactors). Notably, filtration led to an increase in the dominant genus, Zoogloea relative abundance over time within the planktonic community, while remaining relatively constant when unfiltered. At day 23, biofilm communities were more taxonomically and phylogenetically diverse and substantially different from planktonic bacterial communities; however, the biofilm bacterial communities were similar regardless of filtration. These results suggest that although planktonic communities were sensitive to groundwater filtration, bacterial biofilm communities were stable and resistant to filtration. Bioreactors are useful tools in addressing questions pertaining to microbial community assembly and succession. These data provide a first step in understanding how an extrinsic factor, such as a groundwater inoculation and flux of microbial colonizers, impact how microbial communities assemble in environmental systems.« less

Use of in-field bioreactors demonstrate groundwater filtration influences planktonic bacterial community assembly, but not biofilm composition

PubMed Central

Moon, JiWon; Veach, Allison M.; Mosher, Jennifer J.; Wymore, Ann M.; van Nostrand, Joy D.; Zhou, Jizhong; Hazen, Terry C.; Arkin, Adam P.; Elias, Dwayne A.

2018-01-01

Using in-field bioreactors, we investigated the influence of exogenous microorganisms in groundwater planktonic and biofilm microbial communities as part of the Integrated Field Research Challenge (IFRC). After an acclimation period with source groundwater, bioreactors received either filtered (0.22 μM filter) or unfiltered well groundwater in triplicate and communities were tracked routinely for 23 days after filtration was initiated. To address geochemical influences, the planktonic phase was assayed periodically for protein, organic acids, physico-/geochemical measurements and bacterial community (via 16S rRNA gene sequencing), while biofilms (i.e. microbial growth on sediment coupons) were targeted for bacterial community composition at the completion of the experiment (23 d). Based on Bray-Curtis distance, planktonic bacterial community composition varied temporally and between treatments (filtered, unfiltered bioreactors). Notably, filtration led to an increase in the dominant genus, Zoogloea relative abundance over time within the planktonic community, while remaining relatively constant when unfiltered. At day 23, biofilm communities were more taxonomically and phylogenetically diverse and substantially different from planktonic bacterial communities; however, the biofilm bacterial communities were similar regardless of filtration. These results suggest that although planktonic communities were sensitive to groundwater filtration, bacterial biofilm communities were stable and resistant to filtration. Bioreactors are useful tools in addressing questions pertaining to microbial community assembly and succession. These data provide a first step in understanding how an extrinsic factor, such as a groundwater inoculation and flux of microbial colonizers, impact how microbial communities assemble in environmental systems. PMID:29558522

Use of in-field bioreactors demonstrate groundwater filtration influences planktonic bacterial community assembly, but not biofilm composition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Christensen, Geoffrey A.; Moon, Ji Won; Veach, Allison M.

Using in-field bioreactors, we investigated the influence of exogenous microorganisms in groundwater planktonic and biofilm microbial communities as part of the Integrated Field Research Challenge (IFRC). After an acclimation period with source groundwater, bioreactors received either filtered (0.22 μM filter) or unfiltered well groundwater in triplicate and communities were tracked routinely for 23 days after filtration was initiated. To address geochemical influences, the planktonic phase was assayed periodically for protein, organic acids, physico-/geochemical measurements and bacterial community (via 16S rRNA gene sequencing), while biofilms (i.e. microbial growth on sediment coupons) were targeted for bacterial community composition at the completion ofmore » the experiment (23 d). Based on Bray-Curtis distance, planktonic bacterial community composition varied temporally and between treatments (filtered, unfiltered bioreactors). Notably, filtration led to an increase in the dominant genus, Zoogloea relative abundance over time within the planktonic community, while remaining relatively constant when unfiltered. At day 23, biofilm communities were more taxonomically and phylogenetically diverse and substantially different from planktonic bacterial communities; however, the biofilm bacterial communities were similar regardless of filtration. These results suggest that although planktonic communities were sensitive to groundwater filtration, bacterial biofilm communities were stable and resistant to filtration. Bioreactors are useful tools in addressing questions pertaining to microbial community assembly and succession. These data provide a first step in understanding how an extrinsic factor, such as a groundwater inoculation and flux of microbial colonizers, impact how microbial communities assemble in environmental systems.« less

A versatile modular bioreactor platform for Tissue Engineering

PubMed Central

Schuerlein, Sebastian; Schwarz, Thomas; Krziminski, Steffan; Gätzner, Sabine; Hoppensack, Anke; Schwedhelm, Ivo; Schweinlin, Matthias; Walles, Heike

2016-01-01

Abstract Tissue Engineering (TE) bears potential to overcome the persistent shortage of donor organs in transplantation medicine. Additionally, TE products are applied as human test systems in pharmaceutical research to close the gap between animal testing and the administration of drugs to human subjects in clinical trials. However, generating a tissue requires complex culture conditions provided by bioreactors. Currently, the translation of TE technologies into clinical and industrial applications is limited due to a wide range of different tissue‐specific, non‐disposable bioreactor systems. To ensure a high level of standardization, a suitable cost‐effectiveness, and a safe graft production, a generic modular bioreactor platform was developed. Functional modules provide robust control of culture processes, e.g. medium transport, gas exchange, heating, or trapping of floating air bubbles. Characterization revealed improved performance of the modules in comparison to traditional cell culture equipment such as incubators, or peristaltic pumps. By combining the modules, a broad range of culture conditions can be achieved. The novel bioreactor platform allows using disposable components and facilitates tissue culture in closed fluidic systems. By sustaining native carotid arteries, engineering a blood vessel, and generating intestinal tissue models according to a previously published protocol the feasibility and performance of the bioreactor platform was demonstrated. PMID:27492568

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunourous tissues. Here, two High-Aspect Ratio Vessels turn at about 12 rmp to keep breast tissue constructs suspended inside the culture media. Syringes allow scientists to pull for analysis during growth sequences. The tube in the center is a water bubbler that dehumidifies the air to prevent evaporation of the media and thus the appearance of destructive bubbles in the bioreactor.

MONITORING APPROACHES FOR BIOREACTOR LANDFILLS - Report

EPA Science Inventory

Experimental bioreactor landfill operations at operating Municipal Solid Waste (MSW) landfills can be approved under the research development and demonstration (RD&D) provisions of 30CFR 258.4. To provide a basis for consistent data collection for future decision-making in suppor...

Bioreactor design concepts

NASA Technical Reports Server (NTRS)

Bowie, William

1987-01-01

Two parallel lines of work are underway in the bioreactor laboratory. One of the efforts is devoted to the continued development and utilization of a laboratory research system. That system's design is intended to be fluid and dynamic. The sole purpose of such a device is to allow testing and development of equipment concepts and procedures. Some of the results of those processes are discussed. A second effort is designed to produce a flight-like bioreactor contained in a double middeck locker. The result of that effort has been to freeze a particular bioreactor design in order to allow fabrication of the custom parts. The system is expected to be ready for flight in early 1988. However, continued use of the laboratory system will lead to improvements in the space bioreactor. Those improvements can only be integrated after the initial flight series.

Continuous, packed-bed, enzymatic bioreactor production and stability of feruloyl soy glycerides

USDA-ARS?s Scientific Manuscript database

The synthesis of feruloyl soy glycerides was demonstrated on a pilot-scale (1 metric ton/year) in a continuous, four-column series, packed-bed, enzymatic bioreactor (herinafter referred to as the bioreactor). Ethyl ferulate and soybean oil were combined and converted at 3.5 kg/d over Candida antarti...

Earth imaging and scientific observations by SSTI ``Clark'' a NASA technology demonstration spacecraft

NASA Astrophysics Data System (ADS)

Hayduk, Robert J.; Scott, Walter S.; Walberg, Gerald D.; Butts, James J.; Starr, Richard D.

1997-01-01

The Small Satellite Technology Initiative (SSTI) is a National Aeronautics and Space Administration (NASA) program to demonstrate smaller, high technology satellites constructed rapidly and less expensively. Under SSTI, NASA funded the development of ``Clark,'' a high technology demonstration satellite to provide 3-m resolution panchromatic and 15-m resolution multispectral images, as well as collect atmospheric constituent and cosmic x-ray data. The 690-lb. satellite, to be launched in early 1997, will be in a 476 km, circular, sun-synchronous polar orbit. This paper describes the program objectives, the technical characteristics of the sensors and satellite, image processing, archiving and distribution. Data archiving and distribution will be performed by NASA Stennis Space Center and by the EROS Data Center, Sioux Falls, South Dakota, USA.

A comparison of orbitally-shaken and stirred-tank bioreactors: pH modulation and bioreactor type affect CHO cell growth and protein glycosylation.

PubMed

Monteil, Dominique T; Juvet, Valentin; Paz, Jonathan; Moniatte, Marc; Baldi, Lucia; Hacker, David L; Wurm, Florian M

2016-09-01

Orbitally shaken bioreactors (OSRs) support the suspension cultivation of animal cells at volumetric scales up to 200 L and are a potential alternative to stirred-tank bioreactors (STRs) due to their rapid and homogeneous mixing and high oxygen transfer rate. In this study, a Chinese hamster ovary cell line producing a recombinant antibody was cultivated in a 5 L OSR and a 3 L STR, both operated with or without pH control. Effects of bioreactor type and pH control on cell growth and metabolism and on recombinant protein production and glycosylation were determined. In pH-controlled bioreactors, the glucose consumption and lactate production rates were higher relative to cultures grown in bioreactors without pH control. The cell density and viability were higher in the OSRs than in the STRs, either with or without pH control. Volumetric recombinant antibody yields were not affected by the process conditions, and a glycan analysis of the antibody by mass spectrometry did not reveal major process-dependent differences in the galactosylation index. The results demonstrated that OSRs are suitable for recombinant protein production from suspension-adapted animal cells. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1174-1180, 2016. © 2016 American Institute of Chemical Engineers.

Overview of NASA Glenn Aero/Mobile Communications Demonstrations

NASA Technical Reports Server (NTRS)

Brooks, David; Hoder, Doug; Wilkins, Ryan

2004-01-01

The Glenn Research Center at Lewis Field (GRC) has been involved with several other NASA field centers on various networking and RF communications demonstrations and experiments since 1998. These collaborative experiments investigated communications technologies new to aviation, such as wideband Ku satcom, L-band narrowband satcom, and IP (Internet Protocol), using commercial off-the-shelf (COTS) components These technologies can be used to distribute weather and hazard data, air traffic management and airline fleet management information, and passenger cabin Internet service.

Overview of NASA Glenn Aero/Mobile Communication Demonstrations

NASA Technical Reports Server (NTRS)

Brooks, David; Hoder, Doug; Wilkins, Ryan

2004-01-01

The Glenn Research Center at Lewis Field (GRC) has been involved with several other NASA field centers on various networking and RF communications demonstrations and experiments since 1998. These collaborative experiments investigated communications technologies new to aviation, such as wideband Ku satcom, L-band narrowband satcom, and IP (Internet Protocol), using commercial off-the-shelf (COTS) components These technologies can be used to distribute weather and hazard data, air traffic management and airline fleet management information, and passenger cabin Internet service.

Optimizing hydraulic retention times in denitrifying woodchip bioreactors treating recirculating aquaculture system wastewater

USDA-ARS?s Scientific Manuscript database

The performance of wood-based denitrifying bioreactors to treat high-nitrate wastewaters from aquaculture systems has not previously been demonstrated. Four pilot-scale woodchip bioreactors (approximately 1:10 scale) were constructed and operated for 268 d to determine the optimal range of design hy...

A versatile modular bioreactor platform for Tissue Engineering.

PubMed

Schuerlein, Sebastian; Schwarz, Thomas; Krziminski, Steffan; Gätzner, Sabine; Hoppensack, Anke; Schwedhelm, Ivo; Schweinlin, Matthias; Walles, Heike; Hansmann, Jan

2017-02-01

Tissue Engineering (TE) bears potential to overcome the persistent shortage of donor organs in transplantation medicine. Additionally, TE products are applied as human test systems in pharmaceutical research to close the gap between animal testing and the administration of drugs to human subjects in clinical trials. However, generating a tissue requires complex culture conditions provided by bioreactors. Currently, the translation of TE technologies into clinical and industrial applications is limited due to a wide range of different tissue-specific, non-disposable bioreactor systems. To ensure a high level of standardization, a suitable cost-effectiveness, and a safe graft production, a generic modular bioreactor platform was developed. Functional modules provide robust control of culture processes, e.g. medium transport, gas exchange, heating, or trapping of floating air bubbles. Characterization revealed improved performance of the modules in comparison to traditional cell culture equipment such as incubators, or peristaltic pumps. By combining the modules, a broad range of culture conditions can be achieved. The novel bioreactor platform allows using disposable components and facilitates tissue culture in closed fluidic systems. By sustaining native carotid arteries, engineering a blood vessel, and generating intestinal tissue models according to a previously published protocol the feasibility and performance of the bioreactor platform was demonstrated. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Design challenges for space bioreactors

NASA Technical Reports Server (NTRS)

Seshan, P. K.; Petersen, G. R.

1989-01-01

The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived.

Streamlined bioreactor-based production of human cartilage tissues.

PubMed

Tonnarelli, B; Santoro, R; Adelaide Asnaghi, M; Wendt, D

2016-05-27

Engineered tissue grafts have been manufactured using methods based predominantly on traditional labour-intensive manual benchtop techniques. These methods impart significant regulatory and economic challenges, hindering the successful translation of engineered tissue products to the clinic. Alternatively, bioreactor-based production systems have the potential to overcome such limitations. In this work, we present an innovative manufacturing approach to engineer cartilage tissue within a single bioreactor system, starting from freshly isolated human primary chondrocytes, through the generation of cartilaginous tissue grafts. The limited number of primary chondrocytes that can be isolated from a small clinically-sized cartilage biopsy could be seeded and extensively expanded directly within a 3D scaffold in our perfusion bioreactor (5.4 ± 0.9 doublings in 2 weeks), bypassing conventional 2D expansion in flasks. Chondrocytes expanded in 3D scaffolds better maintained a chondrogenic phenotype than chondrocytes expanded on plastic flasks (collagen type II mRNA, 18-fold; Sox-9, 11-fold). After this "3D expansion" phase, bioreactor culture conditions were changed to subsequently support chondrogenic differentiation for two weeks. Engineered tissues based on 3D-expanded chondrocytes were more cartilaginous than tissues generated from chondrocytes previously expanded in flasks. We then demonstrated that this streamlined bioreactor-based process could be adapted to effectively generate up-scaled cartilage grafts in a size with clinical relevance (50 mm diameter). Streamlined and robust tissue engineering processes, as the one described here, may be key for the future manufacturing of grafts for clinical applications, as they facilitate the establishment of compact and closed bioreactor-based production systems, with minimal automation requirements, lower operating costs, and increased compliance to regulatory guidelines.

NASA Systems Autonomy Demonstration Project - Development of Space Station automation technology

NASA Technical Reports Server (NTRS)

Bull, John S.; Brown, Richard; Friedland, Peter; Wong, Carla M.; Bates, William

1987-01-01

A 1984 Congressional expansion of the 1958 National Aeronautics and Space Act mandated that NASA conduct programs, as part of the Space Station program, which will yield the U.S. material benefits, particularly in the areas of advanced automation and robotics systems. Demonstration programs are scheduled for automated systems such as the thermal control, expert system coordination of Station subsystems, and automation of multiple subsystems. The programs focus the R&D efforts and provide a gateway for transfer of technology to industry. The NASA Office of Aeronautics and Space Technology is responsible for directing, funding and evaluating the Systems Autonomy Demonstration Project, which will include simulated interactions between novice personnel and astronauts and several automated, expert subsystems to explore the effectiveness of the man-machine interface being developed. Features and progress on the TEXSYS prototype thermal control system expert system are outlined.

A Modular Instrumentation System for NASA's Habitat Demonstration Unit

NASA Technical Reports Server (NTRS)

Rojdev, Kristina; Kennedy, Kriss; Yim, Hester; Wagner, Raymond S.; Hong, Todd; Studor, George; Delaune, Paul

2010-01-01

NASA's human spaceflight program is focused on developing technologies to expand the reaches of human exploration and science activities beyond low earth orbit. A critical aspect of living in space or on planetary surfaces is habitation, which provides a safe and comfortable space in which humans can live and work. NASA is seeking out the best option for habitation by exploring several different concepts through the Habitat Demonstration Unit (HDU) project. The purpose of this HDU is to develop a fully autonomous habitation system that enables human exploration of space. One critical feature of the HDU project that helps to accomplish its mission of autonomy is the instrumentation system that monitors key subsystems operating within a Habitat configuration. The following paper will discuss previous instrumentation systems used in analog habitat concepts and how the current instrumentation system being implemented on the HDU1-PEM, or pressurized excursion module, is building upon the lessons learned of those previous systems. Additionally, this paper will discuss the benefits and the limitations of implementing a wireless sensor network (WSN) as the basis for data transport in the instrumentation system. Finally, this paper will address the experiences and lessons learned with integration, testing prior to deployment, and field testing at the JSC rock yard. NASA is developing the HDU1-PEM as a step towards a fully autonomous habitation system that enables human exploration of space. To accomplish this purpose, the HDU project is focusing on development, integration, testing, and evaluation of habitation systems. The HDU will be used as a technology pull, testbed, and integration environment in which to advance NASA's understanding of alternative mission architectures, requirements, and operations concepts definition and validation. This project is a multi-year effort. In 2010, the HDU1-PEM will be in a pressurized excursion module configuration, and in 2011 the

NASA's Habitat Demonstration Unit (HDU) Pressurized Excursion Module (PEM) In-Field Demonstration at Desert RATS 2010

NASA Technical Reports Server (NTRS)

Tri, Terry O.; Kennedy, Kriss J.; Toups, Larry; Gill, Tracy R.; Howe, A. Scott

2011-01-01

This paper describes the construction, assembly, subsystem integration, transportation, and field testing operations associated with the Habitat Demonstration Unit (HDU) Pressurized Excursion Module (PEM) and discusses lessons learned. In a one-year period beginning summer 2009, a tightly scheduled design-develop-build process was utilized by a small NASA "tiger team" to produce the functional HDU-PEM prototype in time to participate in the 2010 Desert Research and Technology Studies (Desert RATS) field campaign. The process required the coordination of multiple teams, subcontractors, facility management and safety staff. It also required a well-choreographed material handling and transportation process to deliver the finished product from the NASA-Johnson Space Center facilities to the remote Arizona desert locations of the field test. Significant findings of this paper include the team s greater understanding of the HDU-PEM s many integration issues and the in-field training the team acquired which will enable the implementation of the next-generation of improvements and development of high-fidelity field operations in a harsh environment. The Desert RATS analog environment is being promoted by NASA as an efficient means to design, build, and integrate multiple technologies in a mission architecture context, with the eventual goal of evolving the technologies into robust flight hardware systems. The HDU-PEM in-field demonstration at Desert RATS 2010 provided a validation process for the integration team, which has already begun to retool for the 2011 field tests that require an adapted architecture.

NASA's ATM Technology Demonstration-1: Integrated Concept of Arrival Operations

NASA Technical Reports Server (NTRS)

Baxley, Brian T.; Swenson, Harry N.; Prevot, Thomas; Callantine, Todd J.

2012-01-01

This paper describes operations and procedures envisioned for NASA s Air Traffic Management (ATM) Technology Demonstration #1 (ATD-1). The ATD-1 Concept of Operations (ConOps) demonstration will integrate three NASA technologies to achieve high throughput, fuel-efficient arrival operations into busy terminal airspace. They are Traffic Management Advisor with Terminal Metering (TMA-TM) for precise time-based schedules to the runway and points within the terminal area, Controller-Managed Spacing (CMS) decision support tools for terminal controllers to better manage aircraft delay using speed control, and Flight deck Interval Management (FIM) avionics and flight crew procedures to conduct airborne spacing operations. The ATD-1 concept provides de-conflicted and efficient operations of multiple arrival streams of aircraft, passing through multiple merge points, from top-of-descent (TOD) to touchdown. It also enables aircraft to conduct Optimized Profile Descents (OPDs) from en route altitude to the runway, using primarily speed control to maintain separation and schedule. The ATD-1 project is currently addressing the challenges of integrating the three technologies, and implantation into an operational environment. Goals of the ATD-1 demonstration include increasing the throughput of high-density airports, reducing controller workload, increasing efficiency of arrival operations and the frequency of trajectory-based operations, and promoting aircraft ADS-B equipage.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Isolation of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Isolate of long-term growth human mammary epithelial cells (HMEC) from outgrowth of duct element; cells shown soon after isolation and early in culture in a dish. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Tichmond, NASA/Marshall Space Flight Center (MSFC).

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Breast tissue specimens in traditional sample dishes. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunourous tissues.

NASA Facts: Edison Demonstration of Spacecraft Networks (EDSN) Mission

NASA Technical Reports Server (NTRS)

Ord, Stephen; Yost, Bruce D.; Petro, Andrew J.

2013-01-01

NASA's Edison Demonstration of Smallsat Networks (EDSN) mission will launch and deploy a swarm of 8 cubesats into a loose formation approximately 500 km above Earth. EDSN will develop technology to send multiple, advanced, yet affordable nanosatellites into space with cross-link communications to enable a wide array of scientific, commercial, and academic research. Other goals of the mission include lowering the cost and shortening the development time for future small spacecraft.

Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells

PubMed Central

Osiecki, Michael J.; Michl, Thomas D.; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B.; Griesser, Hans J.; Doran, Michael R.

2015-01-01

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs. PMID:26660475

Packed Bed Bioreactor for the Isolation and Expansion of Placental-Derived Mesenchymal Stromal Cells.

PubMed

Osiecki, Michael J; Michl, Thomas D; Kul Babur, Betul; Kabiri, Mahboubeh; Atkinson, Kerry; Lott, William B; Griesser, Hans J; Doran, Michael R

2015-01-01

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

Aggregation of Culture Expanded Human Mesenchymal Stem Cells in Microcarrier-based Bioreactor.

PubMed

Yuan, Xuegang; Tsai, Ang-Chen; Farrance, Iain; Rowley, Jon; Ma, Teng

2018-03-15

Three-dimensional aggregation of human mesenchymal stem cells (hMSCs) has been used to enhance their therapeutic properties but current fabrication protocols depend on laboratory methods and are not scalable. In this study, we developed thermal responsive poly(N-isopropylacrylamide) grafted microcarriers (PNIPAM-MCs), which supported expansion and thermal detachment of hMSCs at reduced temperature (23.0 °C). hMSCs were cultured on the PNIPAM-MCs in both spinner flask (SF) and PBS Vertical-Wheel (PBS-VW) bioreactors for expansion. At room temperature, hMSCs were detached as small cell sheets, which subsequently self-assembled into 3D hMSC aggregates in PBS-VW bioreactor and remain as single cells in SF bioreactor owing to different hydrodynamic conditions. hMSC aggregates generated from the bioreactor maintained comparable immunomodulation and cytokine secretion properties compared to the ones made from the AggreWell ® . The results of the current study demonstrate the feasibility of scale-up production of hMSC aggregates in the suspension bioreactor using thermal responsive microcarriers for integrated cell expansion and 3D aggregation in a close bioreactor system and highlight the critical role of hydrodynamics in self-assembly of detached hMSC in suspension.

Biofabrication of customized bone grafts by combination of additive manufacturing and bioreactor knowhow.

PubMed

Costa, Pedro F; Vaquette, Cédryck; Baldwin, Jeremy; Chhaya, Mohit; Gomes, Manuela E; Reis, Rui L; Theodoropoulos, Christina; Hutmacher, Dietmar W

2014-09-01

This study reports on an original concept of additive manufacturing for the fabrication of tissue engineered constructs (TEC), offering the possibility of concomitantly manufacturing a customized scaffold and a bioreactor chamber to any size and shape. As a proof of concept towards the development of anatomically relevant TECs, this concept was utilized for the design and fabrication of a highly porous sheep tibia scaffold around which a bioreactor chamber of similar shape was simultaneously built. The morphology of the bioreactor/scaffold device was investigated by micro-computed tomography and scanning electron microscopy confirming the porous architecture of the sheep tibiae as opposed to the non-porous nature of the bioreactor chamber. Additionally, this study demonstrates that both the shape, as well as the inner architecture of the device can significantly impact the perfusion of fluid within the scaffold architecture. Indeed, fluid flow modelling revealed that this was of significant importance for controlling the nutrition flow pattern within the scaffold and the bioreactor chamber, avoiding the formation of stagnant flow regions detrimental for in vitro tissue development. The bioreactor/scaffold device was dynamically seeded with human primary osteoblasts and cultured under bi-directional perfusion for two and six weeks. Primary human osteoblasts were observed homogenously distributed throughout the scaffold, and were viable for the six week culture period. This work demonstrates a novel application for additive manufacturing in the development of scaffolds and bioreactors. Given the intrinsic flexibility of the additive manufacturing technology platform developed, more complex culture systems can be fabricated which would contribute to the advances in customized and patient-specific tissue engineering strategies for a wide range of applications.

NASA Systems Autonomy Demonstration Program - A step toward Space Station automation

NASA Technical Reports Server (NTRS)

Starks, S. A.; Rundus, D.; Erickson, W. K.; Healey, K. J.

1987-01-01

This paper addresses a multiyear NASA program, the Systems Autonomy Demonstration Program (SADP), whose main objectives include the development, integration, and demonstration of automation technology in Space Station flight and ground support systems. The role of automation in the Space Station is reviewed, and the main players in SADP and their roles are described. The core research and technology being promoted by SADP are discussed, and a planned 1988 milestone demonstration of the automated monitoring, operation, and control of a complete mission operations subsystem is addressed.

Space bioreactor: Design/process flow

NASA Technical Reports Server (NTRS)

Cross, John H.

1987-01-01

The design of the space bioreactor stems from three considerations. First, and foremost, it must sustain cells in microgravity. Closely related is the ability to take advantage of the weightlessness and microgravity. Lastly, it should fit into a bioprocess. The design of the space bioreactor is described in view of these considerations. A flow chart of the bioreactor is presented and discussed.

Production of recombinant adeno-associated vectors using two bioreactor configurations at different scales

PubMed Central

Negrete, Alejandro; Kotin, Robert M.

2007-01-01

The conventional methods for producing recombinant adeno-associated virus (rAAV) rely on transient transfection of adherent mammalian cells. To gain acceptance and achieve current good manufacturing process (cGMP) compliance, clinical grade rAAV production process should have the following qualities: simplicity, consistency, cost effectiveness, and scalability. Currently, the only viable method for producing rAAV in large-scale, e.g.≥1016 particles per production run, utilizes Baculovirus Expression Vectors (BEVs) and insect cells suspension cultures. The previously described rAAV production in 40 L culture using a stirred tank bioreactor requires special conditions for implementation and operation not available in all laboratories. Alternatives to producing rAAV in stirred-tank bioreactors are single-use, disposable bioreactors, e.g. Wave™. The disposable bags are purchased pre-sterilized thereby eliminating the need for end-user sterilization and also avoiding cleaning steps between production runs thus facilitating the production process. In this study, rAAV production in stirred tank and Wave™ bioreactors was compared. The working volumes were 10 L and 40 L for the stirred tank bioreactors and 5 L and 20 L for the Wave™ bioreactors. Comparable yields of rAAV, ~2e+13 particles per liter of cell culture were obtained in all volumes and configurations. These results demonstrate that producing rAAV in large scale using BEVs is reproducible, scalable, and independent of the bioreactor configuration. Keywords: adeno-associated vectors; large-scale production; stirred tank bioreactor; wave bioreactor; gene therapy. PMID:17606302

High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.

PubMed

Rameez, Shahid; Mostafa, Sigma S; Miller, Christopher; Shukla, Abhinav A

2014-01-01

Decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. Advanced Microscale Bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant Chinese Hamster Ovary cell line. The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3 L, 15 L, and 200 L stirred tank bioreactors. The processes used in this article involve complex feed formulations, perturbations, and strict process control within the design space, which are in-line with processes used for commercial scale manufacturing of biopharmaceuticals. Changes to important process parameters in ambr™ resulted in predictable cell growth, viability and titer changes, which were in good agreement to data from the conventional larger scale bioreactors. ambr™ was found to successfully reproduce variations in temperature, dissolved oxygen (DO), and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and DO through adjustments to the Proportional and Integral control loop. The data presented here demonstrates the utility of the ambr™ system as a high throughput system for cell culture process development. © 2014 American Institute of Chemical Engineers.

Bioreactor droplets from liposome-stabilized all-aqueous emulsions

NASA Astrophysics Data System (ADS)

Dewey, Daniel C.; Strulson, Christopher A.; Cacace, David N.; Bevilacqua, Philip C.; Keating, Christine D.

2014-08-01

Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ~130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.

Bioreactor droplets from liposome-stabilized all-aqueous emulsions.

PubMed

Dewey, Daniel C; Strulson, Christopher A; Cacace, David N; Bevilacqua, Philip C; Keating, Christine D

2014-08-20

Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ~130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.

Space Bioreactor Science Workshop

NASA Technical Reports Server (NTRS)

Morrison, Dennis R. (Editor)

1987-01-01

The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and a slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells. Applications of microcarrier cultures, development of the first space bioreactor flight system, shear and mixing effects on cells, process control, and methods to monitor cell metabolism and nutrient requirements are among the topics covered.

Microbial Bioreactor Development in the ALS NSCORT

NASA Astrophysics Data System (ADS)

Mitchell, Cary; Whitaker, Dawn; Banks, M. Katherine; Heber, Albert J.; Turco, Ronald F.; Nies, Loring F.; Alleman, James E.; Sharvelle, Sybil E.; Li, Congna; Heller, Megan

The NASA Specialized Center of Research and Training in Advanced Life Support (the ALS NSCORT), a partnership of Alabama A & M, Howard, and Purdue Universities, was established by NASA in 2002 to develop technologies that will reduce the Equivalent System Mass (ESM) of regenerative processes within future space life-support systems. A key focus area of NSCORT research has been the development of efficient microbial bioreactors for treatment of human, crop, and food-process wastes while enabling resource recovery. The approach emphasizes optimizing the energy-saving advantages of hydrolytic enzymes for biomass degradation, with focus on treatment of solid wastes including crop residue, paper, food, and human metabolic wastes, treatment of greywater, cabin air, off-gases from other treatment systems, and habitat condensate. This summary includes important findings from those projects, status of technology development, and recommendations for next steps. The Plant-based Anaerobic-Aerobic Bioreactor-Linked Operation (PAABLO) system was developed to reduce crop residue while generating energy and/or food. Plant residues initially were added directly to the bioreactor, and recalcitrant residue was used as a substrate for growing plants or mushrooms. Subsequently, crop residue was first pretreated with fungi to hydrolyze polymers recalcitrant to bacteria, and leachate from the fungal beds was directed to the anaerobic digester. Exoenzymes from the fungi pre-soften fibrous plant materials, improving recovery of materials that are more easily biodegraded to methane that can be used for energy reclamation. An Autothermal Thermophilic Aerobic Digestion (ATAD) system was developed for biodegradable solid wastes. Objectives were to increase water and nutrient recovery, reduce waste volume, and inactivate pathogens. Operational parameters of the reactor were optimized for degradation and resource recovery while minimizing system requirements and footprint. The start-up behavior

A simple eccentric stirred tank mini-bioreactor: mixing characterization and mammalian cell culture experiments.

PubMed

Bulnes-Abundis, David; Carrillo-Cocom, Leydi M; Aráiz-Hernández, Diana; García-Ulloa, Alfonso; Granados-Pastor, Marisa; Sánchez-Arreola, Pamela B; Murugappan, Gayathree; Alvarez, Mario M

2013-04-01

In industrial practice, stirred tank bioreactors are the most common mammalian cell culture platform. However, research and screening protocols at the laboratory scale (i.e., 5-100 mL) rely primarily on Petri dishes, culture bottles, or Erlenmeyer flasks. There is a clear need for simple-easy to assemble, easy to use, easy to clean-cell culture mini-bioreactors for lab-scale and/or screening applications. Here, we study the mixing performance and culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Laser induced fluorescence (LIF) experiments and computational fluid dynamics (CFD) computations are used to identify the operational conditions required for adequate mixing. Mammalian cell culture experiments were conducted with two different cell models. The specific growth rate and the maximum cell density of Chinese hamster ovary (CHO) cell cultures grown in the mini-bioreactor were comparable to those observed for 6-well culture plates, Erlenmeyer flasks, and 1 L fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded tenfold in suspension conditions using the eccentric mini-bioreactor system. Our results demonstrate good mixing performance and suggest the practicality and adequacy of the proposed mini-bioreactor. Copyright © 2012 Wiley Periodicals, Inc.

Affordable Development and Demonstration of a Small NTR engine and Stage: A Preliminary NASA, DOE, and Industry Assessment

NASA Technical Reports Server (NTRS)

Borowski, S. K.; Sefcik, R. J.; Fittje, J. E.; McCurdy, D. R.; Qualls, A. L.; Schnitzler, B. G; Werner, J.; Weitzberg, A.; Joyner, C. R.

2015-01-01

In FY'11, Nuclear Thermal Propulsion (NTP) was identified as a key propulsion option under the Advanced In-Space Propulsion (AISP) component of NASA's Exploration Technology Development and Demonstration (ETDD) program A strategy was outlined by GRC and NASA HQ that included 2 key elements -"Foundational Technology Development" followed by specific "Technology Demonstration" projects. The "Technology Demonstration "element proposed ground technology demonstration (GTD) testing in the early 2020's, followed by a flight technology demonstration (FTD) mission by approx. 2025. In order to reduce development costs, the demonstration projects would focus on developing a small, low thrust (approx. 7.5 -16.5 klb(f)) engine that utilizes a "common" fuel element design scalable to the higher thrust (approx. 25 klb(f)) engines used in NASA's Mars DRA 5.0 study(NASA-SP-2009-566). Besides reducing development costs and allowing utilization of existing, flight proven engine hard-ware (e.g., hydrogen pumps and nozzles), small, lower thrust ground and flight demonstration engines can validate the technology and offer improved capability -increased payloads and decreased transit times -valued for robotic science missions identified in NASA's Decadal Study.

A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering.

PubMed

Zhang, Zhi-Yong; Teoh, Swee Hin; Teo, Erin Yiling; Khoon Chong, Mark Seow; Shin, Chong Woon; Tien, Foo Toon; Choolani, Mahesh A; Chan, Jerry K Y

2010-11-01

Bioreactors provide a dynamic culture system for efficient exchange of nutrients and mechanical stimulus necessary for the generation of effective tissue engineered bone grafts (TEBG). We have shown that biaxial rotating (BXR) bioreactor-matured human fetal mesenchymal stem cell (hfMSC) mediated-TEBG can heal a rat critical sized femoral defect. However, it is not known whether optimal bioreactors exist for bone TE (BTE) applications. We systematically compared this BXR bioreactor with three most commonly used systems: Spinner Flask (SF), Perfusion and Rotating Wall Vessel (RWV) bioreactors, for their application in BTE. The BXR bioreactor achieved higher levels of cellularity and confluence (1.4-2.5x, p < 0.05) in large 785 mm(3) macroporous scaffolds not achieved in the other bioreactors operating in optimal settings. BXR bioreactor-treated scaffolds experienced earlier and more robust osteogenic differentiation on von Kossa staining, ALP induction (1.2-1.6×, p < 0.01) and calcium deposition (1.3-2.3×, p < 0.01). We developed a Micro CT quantification method which demonstrated homogenous distribution of hfMSC in BXR bioreactor-treated grafts, but not with the other three. BXR bioreactor enabled superior cellular proliferation, spatial distribution and osteogenic induction of hfMSC over other commonly used bioreactors. In addition, we developed and validated a non-invasive quantitative micro CT-based technique for analyzing neo-tissue formation and its spatial distribution within scaffolds. Copyright © 2010 Elsevier Ltd. All rights reserved.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Isolation of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Outgrowth of cells from duct element in upper right corner cultured in a standard dish; most cells spontaneously die during early cell divisions, but a few will establish long-term growth. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Tichmond, NASA/Marshall Space Flight Center (MSFC).

A versatile miniature bioreactor and its application to bioelectrochemistry studies.

PubMed

Kloke, A; Rubenwolf, S; Bücking, C; Gescher, J; Kerzenmacher, S; Zengerle, R; von Stetten, F

2010-08-15

Often, reproducible investigations on bio-microsystems essentially require a flexible but well-defined experimental setup, which in its features corresponds to a bioreactor. We therefore developed a miniature bioreactor with a volume in the range of a few millilitre that is assembled by alternate stacking of individual polycarbonate elements and silicone gaskets. All the necessary supply pipes are incorporated as bore holes or cavities within the individual elements. Their combination allows for a bioreactor assembly that is easily adaptable in size and functionality to experimental demands. It allows for controlling oxygen transfer as well as the monitoring of dissolved oxygen concentration and pH-value. The system provides access for media exchange or sterile sampling. A mass transfer coefficient for oxygen (k(L)a) of 4.3x10(-3) s(-1) at a flow rate of only 15 ml min(-1) and a mixing time of 1.5s at a flow rate of 11 ml min(-1) were observed for the modular bioreactor. Single reactor chambers can be interconnected via ion-conductive membranes to form a two-chamber test setup for investigations on electrochemical systems such as fuel cells or sensors. The versatile applicability of this modular and flexible bioreactor was demonstrated by recording a growth curve of Escherichia coli (including monitoring of pH and oxygen) saturation, and also as by two bioelectrochemical experiments. In the first electrochemical experiment the use of the bioreactor enabled a direct comparison of electrode materials for a laccase-catalyzed oxygen reduction electrode. In a second experiment, the bioreactor was utilized to characterize the influence of outer membrane cytochromes on the performance of Shewanella oneidensis in a microbial fuel cell. Copyright 2010 Elsevier B.V. All rights reserved.

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Dr. Robert Richmond extracts breast cell tissue from one of two liquid nitrogen dewars. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunourous tissues.

Mechanobiologic Research in a Microgravity Environment Bioreactor

NASA Astrophysics Data System (ADS)

Guidi, A.; Dubini, G.; Tominetti, F.; Raimondi, M.

A current problem in tissue culturing technology is the unavailability of an effective Bioreactor for the in vitro cultivation of cells and explants. It has, in fact, proved extremely difficult to promote the high-density three-dimensional in vitro growth of human tissues that have been removed from the body and deprived of their normal in vivo vascular sources of nutrients and gas exchange. A variety of tissue explants can be maintained for a short period of time on a supportive collagen matrix surrounded by culture medium. But this system provides only limited mass transfer of nutrients and wastes through the tissue, and gravity-induced sedimentation prevents complete three- dimensional cell-cell and cell-matrix interactions. Several devices presently on the market have been used with only limited success since each has limitations, which restrict usefulness and versatility. Further, no Bioreactor or culture vessel is known that will allow for unimpeded growth of three dimensional cellular aggregates or tissue. Extensive research on the effect of mechanical stimuli on cell metabolism suggests that tissues may respond to mechanical stimulation via loading-induced flow of the interstitial fluids. During the culture, cells are subject to a flow of culture medium. Flow properties such as flow field, flow regime (e.g. turbulent or laminar), flow pattern (e.g. circular), entity and distribution of the shear stress acting on the cells greatly influence fundamental aspects of cell function, such as regulation and gene expression. This has been demonstrated for endothelial cells and significant research efforts are underway to elucidate these mechanisms in various other biological systems. Local fluid dynamics is also responsible of the mass transfer of nutrients and catabolites as well as oxygenation through the tissue. Most of the attempts to culture tissue-engineered constructs in vitro have utilized either stationary cultures or systems generating relatively small

Bioreactor cultivation enhances NTEB formation and differentiation of NTES cells into cardiomyocytes.

PubMed

Lü, Shuanghong; Liu, Sheng; He, Wenjun; Duan, Cuimi; Li, Yanmin; Liu, Zhiqiang; Zhang, Ye; Hao, Tong; Wang, Yanmeng; Li, Dexue; Wang, Changyong; Gao, Shaorong

2008-09-01

Autogenic embryonic stem cells established from somatic cell nuclear transfer (SCNT) embryos have been proposed as unlimited cell sources for cell transplantation-based treatment of many genetic and degenerative diseases, which can eliminate the immune rejection that occurs after transplantation. In the present study, pluripotent nuclear transfer ES (NTES) cell lines were successfully established from different strains of mice. One NTES cell line, NT1, with capacity of germline transmission, was used to investigate in vitro differentiation into cardiomyocytes. To optimize differentiation conditions for mass production of embryoid bodies (NTEBs) from NTES cells, a slow-turning lateral vessel (STLV) rotating bioreactor was used for culturing the NTES cells to produce NTEBs compared with a conventional static cultivation method. Our results demonstrated that the NTEBs formed in STLV bioreactor were more uniform in size, and no large necrotic centers with most of the cells in NTEBs were viable. Differentiation of the NTEBs formed in both the STLV bioreactor and static culture into cardiomyocytes was induced by ascorbic acid, and the results demonstrated that STLV-produced NTEBs differentiated into cardiomyocytes more efficiently. Taken together, our results suggested that STLV bioreactor provided a more ideal culture condition, which can facilitate the formation of better quality NTEBs and differentiation into cardiomyocytes more efficiently in vitro.

NASA systems autonomy demonstration project: Advanced automation demonstration of Space Station Freedom thermal control system

NASA Technical Reports Server (NTRS)

Dominick, Jeffrey; Bull, John; Healey, Kathleen J.

1990-01-01

The NASA Systems Autonomy Demonstration Project (SADP) was initiated in response to Congressional interest in Space station automation technology demonstration. The SADP is a joint cooperative effort between Ames Research Center (ARC) and Johnson Space Center (JSC) to demonstrate advanced automation technology feasibility using the Space Station Freedom Thermal Control System (TCS) test bed. A model-based expert system and its operator interface were developed by knowledge engineers, AI researchers, and human factors researchers at ARC working with the domain experts and system integration engineers at JSC. Its target application is a prototype heat acquisition and transport subsystem of a space station TCS. The demonstration is scheduled to be conducted at JSC in August, 1989. The demonstration will consist of a detailed test of the ability of the Thermal Expert System to conduct real time normal operations (start-up, set point changes, shut-down) and to conduct fault detection, isolation, and recovery (FDIR) on the test article. The FDIR will be conducted by injecting ten component level failures that will manifest themselves as seven different system level faults. Here, the SADP goals, are described as well as the Thermal Control Expert System that has been developed for demonstration.

Tunable osteogenic differentiation of hMPCs in tubular perfusion system bioreactor.

PubMed

Nguyen, Bao-Ngoc B; Ko, Henry; Fisher, John P

2016-08-01

The use of bioreactors for bone tissue engineering has been widely investigated. While the benefits of shear stress on osteogenic differentiation are well known, the underlying effects of dynamic culture on subpopulations within a bioreactor are less evident. In this work, we explore the influence of applied flow in the tubular perfusion system (TPS) bioreactor on the osteogenic differentiation of human mesenchymal progenitor cells (hMPCs), specifically analyzing the effects of axial position along the growth chamber. TPS bioreactor experiments conducted with unidirectional flow demonstrated enhanced expression of osteogenic markers in cells cultured downstream from the inlet flow. We utilized computational fluid dynamic modeling to confirm uniform shear stress distribution on the surface of the scaffolds and along the length of the growth chamber. The concept of paracrine signaling between cell populations was validated with the use of alternating flow, which diminished the differences in osteogenic differentiation between cells cultured at the inlet and outlet of the growth chamber. After the addition of controlled release of bone morphogenic protein-2 (BMP-2) into the system, osteogenic differentiation among subpopulations along the growth chamber was augmented, yet remained homogenous. These results allow for greater understanding of axial bioreactor cultures, their microenvironment, and how well-established parameters of osteogenic differentiation affect bone tissue development. With this work, we have demonstrated the capability of tuning osteogenic differentiation of hMPCs through the application of fluid flow and the addition of exogenous growth factors. Such precise control allows for the culture of distinct subpopulation within one dynamic system for the use of complex engineered tissue constructs. Biotechnol. Bioeng. 2016;113: 1805-1813. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Use of microgravity bioreactors for development of an in vitro rat salivary gland cell culture model

NASA Technical Reports Server (NTRS)

Lewis, M. L.; Moriarity, D. M.; Campbell, P. S.

1993-01-01

During development, salivary gland (SG) cells both secrete factors which modulate cellular behavior and express specific hormone receptors. Whether SG cell growth is modulated by an autocrine epidermal growth factor (EGF) receptor-mediated signal transduction pathway is not clearly understood. SG tissue is the synthesis site for functionally distinct products including growth factors, digestive enzymes, and homeostasis maintaining factors. Historically, SG cells have proven difficult to grow and may be only maintained as limited three-dimensional ductal-type structures in collagen gels or on reconstituted basement membrane gels. A novel approach to establishing primary rat SG cultures is use of microgravity bioreactors originally designed by NASA as low-shear culture systems for predicting cell growth and differentiation in the microgravity environment of space. These completely fluid-filled bioreactors, which are oriented horizontally and rotate, have proven advantageous for Earth-based culture of three-dimensional cell assemblies, tissue-like aggregates, and glandular structures. Use of microgravity bioreactors for establishing in vitro models to investigate steroid-mediated secretion of EGF by normal SG cells may also prove useful for the investigation of cancer and other salivary gland disorders. These microgravity bioreactors promise challenging opportunities for future applications in basic and applied cell research.

An update to space biomedical research: tissue engineering in microgravity bioreactors.

PubMed

Barzegari, Abolfazl; Saei, Amir Ata

2012-01-01

The severe need for constructing replacement tissues in organ transplanta-tion has necessitated the development of tissue engineering approaches and bioreactors that can bring these approaches to reality. The inherent limitations of conventional bioreactors in generating realistic tissue constructs led to the devise of the microgravity tissue engineering that uses Rotating Wall Vessel (RWV) bioreactors initially developed by NASA. In this review article, we intend to highlight some major advances and accomplishments in the rapidly-growing field of tissue engineering that could not be achieved without using microgravity. Research is now focused on assembly of 3 dimensional (3D) tissue fragments from various cell types in human body such as chon-drocytes, osteoblasts, embryonic and mesenchymal stem cells, hepatocytes and pancreas islet cells. Hepatocytes cultured under microgravity are now being used in extracorporeal bioartificial liver devices. Tissue constructs can be used not only in organ replacement therapy, but also in pharmaco-toxicology and food safety assessment. 3D models of vari-ous cancers may be used in studying cancer development and biology or in high-throughput screening of anticancer drug candidates. Finally, 3D heterogeneous assemblies from cancer/immune cells provide models for immunotherapy of cancer. Tissue engineering in (simulated) microgravity has been one of the stunning impacts of space research on biomedical sciences and their applications on earth.

Lessons Learned from NASA UAV Science Demonstration Program Missions

NASA Technical Reports Server (NTRS)

Wegener, Steven S.; Schoenung, Susan M.

2003-01-01

During the summer of 2002, two airborne missions were flown as part of a NASA Earth Science Enterprise program to demonstrate the use of uninhabited aerial vehicles (UAVs) to perform earth science. One mission, the Altus Cumulus Electrification Study (ACES), successfully measured lightning storms in the vicinity of Key West, Florida, during storm season using a high-altitude Altus(TM) UAV. In the other, a solar-powered UAV, the Pathfinder Plus, flew a high-resolution imaging mission over coffee fields in Kauai, Hawaii, to help guide the harvest.

Modeling energy consumption in membrane bioreactors for wastewater treatment in north Africa.

PubMed

Skouterisl, George; Arnot, Tom C; Jraou, Mouna; Feki, Firas; Sayadi, Sami

2014-03-01

Two pilot-scale membrane bioreactors were operated alongside a full-sized activated sludge plant in Tunisia in order to compare specific energy demand and treated water quality. Energy consumption rates were measured for the complete membrane bioreactor systems and for their different components. Specific energy demand was measured for the systems and compared with the activated sludge plant, which operated at around 3 kWh m(-3). A model was developed for each membrane bioreactor based on both dynamic and steady-state mass balances, microbial kinetics and stoichiometry, and energy balance. Energy consumption was evaluated as a function of mixed-liquor suspended solids concentration, net permeate fluxes, and the resultant treated water quality. This work demonstrates the potential for using membrane bioreactors in decentralised domestic water treatment in North Africa, at energy consumption levels similar or lower than conventional activated sludge systems, with the added benefit of producing treated water suitable for unrestricted crop irrigation.

Unique differentiation profile of mouse embryonic stem cells in rotary and stirred tank bioreactors.

PubMed

Fridley, Krista M; Fernandez, Irina; Li, Mon-Tzu Alice; Kettlewell, Robert B; Roy, Krishnendu

2010-11-01

Embryonic stem (ES)-cell-derived lineage-specific stem cells, for example, hematopoietic stem cells, could provide a potentially unlimited source for transplantable cells, especially for cell-based therapies. However, reproducible methods must be developed to maximize and scale-up ES cell differentiation to produce clinically relevant numbers of therapeutic cells. Bioreactor-based dynamic culture conditions are amenable to large-scale cell production, but few studies have evaluated how various bioreactor types and culture parameters influence ES cell differentiation, especially hematopoiesis. Our results indicate that cell seeding density and bioreactor speed significantly affect embryoid body formation and subsequent generation of hematopoietic stem and progenitor cells in both stirred tank (spinner flask) and rotary microgravity (Synthecon™) type bioreactors. In general, high percentages of hematopoietic stem and progenitor cells were generated in both bioreactors, especially at high cell densities. In addition, Synthecon bioreactors produced more sca-1(+) progenitors and spinner flasks generated more c-Kit(+) progenitors, demonstrating their unique differentiation profiles. cDNA microarray analysis of genes involved in pluripotency, germ layer formation, and hematopoietic differentiation showed that on day 7 of differentiation, embryoid bodies from both bioreactors consisted of all three germ layers of embryonic development. However, unique gene expression profiles were observed in the two bioreactors; for example, expression of specific hematopoietic genes were significantly more upregulated in the Synthecon cultures than in spinner flasks. We conclude that bioreactor type and culture parameters can be used to control ES cell differentiation, enhance unique progenitor cell populations, and provide means for large-scale production of transplantable therapeutic cells.

PRACTICE REVIEW OF FIVE BIOREACTOR/RECIRCULATION LANDFILLS

EPA Science Inventory

Six bioreactor landfills were analyzed to provide a perspective of current practice and technical issues that differentiate bioreactor landfills from conventional landfills. Five of the bioreactor landfills were anaerobic and one was aerated. In one case, nearly identical cells e...

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Isolation of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue. Same long-term growth human mammary epithelial cells (HMEC), but after 3 weeks in concinuous culture. Note attempts to reform duct elements, but this time in two dimensions in a dish rather that in three demensions in tissue. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Tichmond, NASA/Marshall Space Flight Center (MSFC).

Biotechnology

NASA Image and Video Library

2002-07-02

Leland W. K. Chung (left), Director, Molecular Urology Therapeutics Program at the Winship Cancer Institute at Emory University, is principal investigator for the NASA bioreactor demonstration system (BDS-05). With him is Dr. Jun Shu, an assistant professor of Orthopedics Surgery from Kuming Medical University China. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: Emory University.

EMERGING TECHNOLOGY SUMMARY: PILOT-SCALE DEMONSTRATION OF A TWO-STAGE METHANOTROPHIC BIOREACTOR FOR BIODEGRADATION OF TRICHLOROETHENE IN GROUNDWATER

EPA Science Inventory

BioTrol, Inc., developed a two-stage, methanotrophic, bioreactor system for remediation of water contaminated with trichloroethylene (TCE) and other chlorinated, volatile, aliphatic hydrocarbons. The first stage was a suspended-growth culture vessel with a bubbleless methane tran...

Potential use of the bioreactor to determine effects of microgravity and other environmental parameters on growth of hybridoma cells

NASA Technical Reports Server (NTRS)

Ley, Kenneth D.

1987-01-01

It is argued that the bioreactor being developed at NASA will allow researchers to determine the optimal conditions (e.g., pH, O sub 2, CO sub 2, nutrients) for growth of hybridoma cells, and to determine whether cell growth and antibody production are enhanced in the microgravity of space.

Analysis of the efficiency of recombinant Escherichia coli strain cultivation in a gas-vortex bioreactor.

PubMed

Savelyeva, Anna V; Nemudraya, Anna A; Podgornyi, Vladimir F; Laburkina, Nadezhda V; Ramazanov, Yuriy A; Repkov, Andrey P; Kuligina, Elena V; Richter, Vladimir A

2017-09-01

The levels of aeration and mass transfer are critical parameters required for an efficient aerobic bioprocess, and directly depend on the design features of exploited bioreactors. A novel apparatus, using gas vortex for aeration and mass transfer processes, was constructed in the Center of Vortex Technologies (Novosibirsk, Russia). In this paper, we compared the efficiency of recombinant Escherichia coli strain cultivation using novel gas-vortex technology with conventional bioprocess technologies such as shake flasks and bioreactors with mechanical stirrers. We demonstrated that the system of aeration and agitation used in gas-vortex bioreactors provides 3.6 times higher volumetric oxygen transfer coefficient in comparison with mechanical bioreactor. The use of gas-vortex bioreactor for recombinant E. coli strain cultivation allows to increase the efficiency of target protein expression at 2.2 times for BL21(DE3)/pFK2 strain and at 3.5 times for auxotrophic C600/pRT strain (in comparison with stirred bioreactor). © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Optimising Microbial Growth with a Bench-Top Bioreactor

ERIC Educational Resources Information Center

Baker, A. M. R.; Borin, S. L.; Chooi, K. P.; Huang, S. S.; Newgas, A. J. S.; Sodagar, D.; Ziegler, C. A.; Chan, G. H. T.; Walsh, K. A. P.

2006-01-01

The effects of impeller size, agitation and aeration on the rate of yeast growth were investigated using bench-top bioreactors. This exercise, carried out over a six-month period, served as an effective demonstration of the importance of different operating parameters on cell growth and provided a means of determining the optimisation conditions…

Mercury Ion Clock for a NASA Technology Demonstration Mission.

PubMed

Tjoelker, Robert L; Prestage, John D; Burt, Eric A; Chen, Pin; Chong, Yong J; Chung, Sang K; Diener, William; Ely, Todd; Enzer, Daphna G; Mojaradi, Hadi; Okino, Clay; Pauken, Mike; Robison, David; Swenson, Bradford L; Tucker, Blake; Wang, Rabi

2016-07-01

There are many different atomic frequency standard technologies but only few meet the demanding performance, reliability, size, mass, and power constraints required for space operation. The Jet Propulsion Laboratory is developing a linear ion-trap-based mercury ion clock, referred to as DSAC (Deep-Space Atomic Clock) under NASA's Technology Demonstration Mission program. This clock is expected to provide a new capability with broad application to space-based navigation and science. A one-year flight demonstration is planned as a hosted payload following an early 2017 launch. This first-generation mercury ion clock for space demonstration has a volume, mass, and power of 17 L, 16 kg, and 47 W, respectively, with further reductions planned for follow-on applications. Clock performance with a signal-to-noise ratio (SNR)*Q limited stability of 1.5E-13/τ(1/2) has been observed and a fractional frequency stability of 2E-15 at one day measured (no drift removed). Such a space-based stability enables autonomous timekeeping of with a technology capable of even higher stability, if desired. To date, the demonstration clock has been successfully subjected to mechanical vibration testing at the 14 grms level, thermal-vacuum operation over a range of 42(°)C, and electromagnetic susceptibility tests.

Diagram of Cell to Cell Communication

NASA Technical Reports Server (NTRS)

2002-01-01

Diagram depicts the importance of cell-cell communication as central to the understanding of cancer growth and progression, the focus of the NASA bioreactor demonstration system (BDS-05) investigation. Microgravity studies will allow us to unravel the signaling and communication between these cells with the host and potential development of therapies for the treatment of cancer metastasis. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: Emory University.

Biotechnology

NASA Image and Video Library

2002-07-02

Diagram depicts the importance of cell-cell communication as central to the understanding of cancer growth and progression, the focus of the NASA bioreactor demonstration system (BDS-05) investigation. Microgravity studies will allow us to unravel the signaling and communication between these cells with the host and potential development of therapies for the treatment of cancer metastasis. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: Emory University.

TREATMENT OF MUNICIPAL WASTEWATERS BY THE FLUIDIZED BED BIOREACTOR PROCESS

EPA Science Inventory

A 2-year, large-scale pilot investigation was conducted at the City of Newburgh Water Pollution Control Plant, Newburgh, NY, to demonstrate the application of the fluidized bed bioreactor process to the treatment of municipal wastewaters. The experimental effort investigated the ...

Bioreactor design for tendon/ligament engineering.

PubMed

Wang, Tao; Gardiner, Bruce S; Lin, Zhen; Rubenson, Jonas; Kirk, Thomas B; Wang, Allan; Xu, Jiake; Smith, David W; Lloyd, David G; Zheng, Ming H

2013-04-01

Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a suitable culture environment, which mimics the dynamics of the in vivo environment for tendon/ligament maturation. For clinical settings, bioreactors also have the advantages of less-contamination risk, high reproducibility of cell propagation by minimizing manual operation, and a consistent end product. In this review, we identify the key components, design preferences, and criteria that are required for the development of an ideal bioreactor for engineering tendons and ligaments.

Advanced bioreactors for enhanced production of chemicals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Davison, B.H.; Scott, C.D.

1993-06-01

A variety of advanced bioreactors are being developed to improve production of fuels, solvents, organic acids and other fermentation products. One key approach is immobilization of the biocatalyst leading to increased rates and yields. In addition, there are processes for simultaneous fermentation and separation to further increase production by the removal of an inhibitory product. For example, ethanol productivity in immobilized-cell fluidized-bed bioreactors (FBRs) can increase more than tenfold with 99% conversion and near stoichiometric yields. Two modified FBR configurations offer further improvements by removing the inhibitory product directly from the continuous fermentation. One involves the addition and removal ofmore » solid adsorbent particles to the FBR. This process was demonstrated with the production of lactic acid by immobilized Lactobacillus. The second uses an immiscible organic extractant in the FBR. This increased total butanol yields in the anaerobic acetone-butanol fermentation by Clostridium acetobutylicum.« less

Designing electrical stimulated bioreactors for nerve tissue engineering

NASA Astrophysics Data System (ADS)

Sagita, Ignasius Dwi; Whulanza, Yudan; Dhelika, Radon; Nurhadi, Ibrahim

2018-02-01

Bioreactor provides a biomimetic ecosystem that is able to culture cells in a physically controlled system. In general, the controlled-parameters are temperature, pH, fluid flow, nutrition flow, etc. In this study, we develop a bioreactor that specifically targeted to culture neural stem cells. This bioreactor could overcome some limitations of conventional culture technology, such as petri dish, by providing specific range of observation area and a uniform treatment. Moreover, the microfluidic bioreactor, which is a small-controlled environment, is able to observe as small number of cells as possible. A perfusion flow is applied to mimic the physiological environment in human body. Additionally, this bioreactor also provides an electrical stimulation which is needed by neural stem cells. In conclusion, we found the correlation between the induced shear stress with geometric parameters of the bioreactor. Ultimately, this system shall be used to observe the interaction between stimulation and cell growth.

Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes.

PubMed

Shafa, Mehdi; Krawetz, Roman; Zhang, Yuan; Rattner, Jerome B; Godollei, Anna; Duff, Henry J; Rancourt, Derrick E

2011-12-14

Embryonic stem cells (ESCs) can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs). However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Murine D3-MHC-neo(r) ESCs formed embyroid bodies (EBs) and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin) was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC differentiation protocols within suspension bioreactors demands a

RWPV bioreactor mass transport: earth-based and in microgravity

NASA Technical Reports Server (NTRS)

Begley, Cynthia M.; Kleis, Stanley J.

2002-01-01

Mass transport and mixing of perfused scalar quantities in the NASA Rotating Wall Perfused Vessel bioreactor are studied using numerical models of the flow field and scalar concentration field. Operating conditions typical of both microgravity and ground-based cell cultures are studied to determine the expected vessel performance for both flight and ground-based control experiments. Results are presented for the transport of oxygen with cell densities and consumption rates typical of colon cancer cells cultured in the RWPV. The transport and mixing characteristics are first investigated with a step change in the perfusion inlet concentration by computing the time histories of the time to exceed 10% inlet concentration. The effects of a uniform cell utilization rate are then investigated with time histories of the outlet concentration, volume average concentration, and volume fraction starved. It is found that the operating conditions used in microgravity produce results that are quite different then those for ground-based conditions. Mixing times for microgravity conditions are significantly shorter than those for ground-based operation. Increasing the differential rotation rates (microgravity) increases the mixing and transport, while increasing the mean rotation rate (ground-based) suppresses both. Increasing perfusion rates enhances mass transport for both microgravity and ground-based cases, however, for the present range of operating conditions, above 5-10 cc/min there are diminishing returns as much of the inlet fluid is transported directly to the perfusion exit. The results show that exit concentration is not a good indicator of the concentration distributions in the vessel. In microgravity conditions, the NASA RWPV bioreactor with the viscous pump has been shown to provide an environment that is well mixed. Even when operated near the theoretical minimum perfusion rates, only a small fraction of the volume provides less than the required oxygen levels

RWPV bioreactor mass transport: earth-based and in microgravity.

PubMed

Begley, Cynthia M; Kleis, Stanley J

2002-11-20

Mass transport and mixing of perfused scalar quantities in the NASA Rotating Wall Perfused Vessel bioreactor are studied using numerical models of the flow field and scalar concentration field. Operating conditions typical of both microgravity and ground-based cell cultures are studied to determine the expected vessel performance for both flight and ground-based control experiments. Results are presented for the transport of oxygen with cell densities and consumption rates typical of colon cancer cells cultured in the RWPV. The transport and mixing characteristics are first investigated with a step change in the perfusion inlet concentration by computing the time histories of the time to exceed 10% inlet concentration. The effects of a uniform cell utilization rate are then investigated with time histories of the outlet concentration, volume average concentration, and volume fraction starved. It is found that the operating conditions used in microgravity produce results that are quite different then those for ground-based conditions. Mixing times for microgravity conditions are significantly shorter than those for ground-based operation. Increasing the differential rotation rates (microgravity) increases the mixing and transport, while increasing the mean rotation rate (ground-based) suppresses both. Increasing perfusion rates enhances mass transport for both microgravity and ground-based cases, however, for the present range of operating conditions, above 5-10 cc/min there are diminishing returns as much of the inlet fluid is transported directly to the perfusion exit. The results show that exit concentration is not a good indicator of the concentration distributions in the vessel. In microgravity conditions, the NASA RWPV bioreactor with the viscous pump has been shown to provide an environment that is well mixed. Even when operated near the theoretical minimum perfusion rates, only a small fraction of the volume provides less than the required oxygen levels

A novel milliliter-scale chemostat system for parallel cultivation of microorganisms in stirred-tank bioreactors.

PubMed

Schmideder, Andreas; Severin, Timm Steffen; Cremer, Johannes Heinrich; Weuster-Botz, Dirk

2015-09-20

A pH-controlled parallel stirred-tank bioreactor system was modified for parallel continuous cultivation on a 10 mL-scale by connecting multichannel peristaltic pumps for feeding and medium removal with micro-pipes (250 μm inner diameter). Parallel chemostat processes with Escherichia coli as an example showed high reproducibility with regard to culture volume and flow rates as well as dry cell weight, dissolved oxygen concentration and pH control at steady states (n=8, coefficient of variation <5%). Reliable estimation of kinetic growth parameters of E. coli was easily achieved within one parallel experiment by preselecting ten different steady states. Scalability of milliliter-scale steady state results was demonstrated by chemostat studies with a stirred-tank bioreactor on a liter-scale. Thus, parallel and continuously operated stirred-tank bioreactors on a milliliter-scale facilitate timesaving and cost reducing steady state studies with microorganisms. The applied continuous bioreactor system overcomes the drawbacks of existing miniaturized bioreactors, like poor mass transfer and insufficient process control. Copyright © 2015 Elsevier B.V. All rights reserved.

Process technology of luwak coffee through bioreactor utilization

NASA Astrophysics Data System (ADS)

Hadipernata, M.; Nugraha, S.

2018-01-01

Indonesia has an advantage in producing exotic coffee that is Luwak coffee. Luwak coffee is produced from the fermentation process in digestion of civet. Luwak coffee production is still limited due to the difficulty level in the use of civet animals as the only medium of Luwak coffee making. The research was conducted by developing technology of luwak coffee production through bioreactor utilization and addition the bacteria isolate from gastric of civet. The process conditions in the bioreactor which include temperature, pH, and bacteria isolate of civet are adjusted to the process that occurs in civet digestion, including peristaltic movement on the stomach and small intestine of the civet will be replaced by the use of propellers that rotate on the bioreactor. The result of research showed that proximat analysis data of artificial/bioreactor luwak coffee did not significant different with original luwak coffee. However, the original luwak coffee has higher content of caffeine compared to bioreactor luwak coffee. Based on the cuping test the bioreactor luwak coffee has a value of 84.375, while the original luwak coffee is 84.875. As the result, bioreactor luwak coffee has excellent taste that similiar with original luwak coffee taste.

Bioreactor Design for Tendon/Ligament Engineering

PubMed Central

Wang, Tao; Gardiner, Bruce S.; Lin, Zhen; Rubenson, Jonas; Kirk, Thomas B.; Wang, Allan; Xu, Jiake

2013-01-01

Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a suitable culture environment, which mimics the dynamics of the in vivo environment for tendon/ligament maturation. For clinical settings, bioreactors also have the advantages of less-contamination risk, high reproducibility of cell propagation by minimizing manual operation, and a consistent end product. In this review, we identify the key components, design preferences, and criteria that are required for the development of an ideal bioreactor for engineering tendons and ligaments. PMID:23072472

The Lunar Laser Communication Demonstration: NASA's First Step Toward Very High Data Rate Support of Science and Exploration Missions

NASA Astrophysics Data System (ADS)

Boroson, Don M.; Robinson, Bryan S.

2014-12-01

Future NASA missions for both Science and Exploration will have needs for much higher data rates than are presently available, even with NASA's highly-capable Space- and Deep-Space Networks. As a first step towards this end, for one month in late 2013, NASA's Lunar Laser Communication Demonstration (LLCD) successfully demonstrated for the first time high-rate duplex laser communications between a satellite in lunar orbit, the Lunar Atmosphere and Dust Environment Explorer (LADEE), and multiple ground stations on the Earth. It constituted the longest-range laser communication link ever built and demonstrated the highest communication data rates ever achieved to or from the Moon.

Schisandra lignans production regulated by different bioreactor type.

PubMed

Szopa, Agnieszka; Kokotkiewicz, Adam; Luczkiewicz, Maria; Ekiert, Halina

2017-04-10

Schisandra chinensis (Chinese magnolia vine) is a rich source of therapeutically relevant dibenzocyclooctadiene lignans with anticancer, immunostimulant and hepatoprotective activities. In this work, shoot cultures of S. chinensis were grown in different types of bioreactors with the aim to select a system suitable for the large scale in vitro production of schisandra lignans. The cultures were maintained in Murashige-Skoog (MS) medium supplemented with 3mg/l 6-benzylaminopurine (BA) and 1mg/l 1-naphthaleneacetic acid (NAA). Five bioreactors differing with respect to cultivation mode were tested: two liquid-phase systems (baloon-type bioreactor and bubble-column bioreactor with biomass immobilization), the gas-phase spray bioreactor and two commercially available temporary immersion systems: RITA ® and Plantform. The experiments were run for 30 and 60 days in batch mode. The harvested shoots were evaluated for growth and lignan content determined by LC-DAD and LC-DAD-ESI-MS. Of the tested bioreactors, temporary immersion systems provided the best results with respect to biomass production and lignan accumulation: RITA ® bioreactor yielded 17.86g/l (dry weight) during 60 day growth period whereas shoots grown for 30 days in Plantform bioreactor contained the highest amount of lignans (546.98mg/100g dry weight), with schisandrin, deoxyschisandrin and gomisin A as the major constituents (118.59, 77.66 and 67.86mg/100g dry weight, respectively). Copyright © 2017 Elsevier B.V. All rights reserved.

Bioreactor-induced mesenchymal progenitor cell differentiation and elastic fiber assembly in engineered vascular tissues.

PubMed

Lin, Shigang; Mequanint, Kibret

2017-09-01

In vitro maturation of engineered vascular tissues (EVT) requires the appropriate incorporation of smooth muscle cells (SMC) and extracellular matrix (ECM) components similar to native arteries. To this end, the aim of the current study was to fabricate 4mm inner diameter vascular tissues using mesenchymal progenitor cells seeded into tubular scaffolds. A dual-pump bioreactor operating either in perfusion or pulsatile perfusion mode was used to generate physiological-like stimuli to promote progenitor cell differentiation, extracellular elastin production, and tissue maturation. Our data demonstrated that pulsatile forces and perfusion of 3D tubular constructs from both the lumenal and ablumenal sides with culture media significantly improved tissue assembly, effectively inducing mesenchymal progenitor cell differentiation to SMCs with contemporaneous elastin production. With bioreactor cultivation, progenitor cells differentiated toward smooth muscle lineage characterized by the expression of smooth muscle (SM)-specific markers smooth muscle alpha actin (SM-α-actin) and smooth muscle myosin heavy chain (SM-MHC). More importantly, pulsatile perfusion bioreactor cultivation enhanced the synthesis of tropoelastin and its extracellular cross-linking into elastic fiber compared with static culture controls. Taken together, the current study demonstrated progenitor cell differentiation and vascular tissue assembly, and provides insights into elastin synthesis and assembly to fibers. Incorporation of elastin into engineered vascular tissues represents a critical design goal for both mechanical and biological functions. In the present study, we seeded porous tubular scaffolds with multipotent mesenchymal progenitor cells and cultured in dual-pump pulsatile perfusion bioreactor. Physiological-like stimuli generated by bioreactor not only induced mesenchymal progenitor cell differentiation to vascular smooth muscle lineage but also actively promoted elastin synthesis and

Clinostats and bioreactors.

PubMed

Klaus, D M

2001-06-01

The environment created on Earth within a clinostat or Rotating Wall Vessel (RWV) bioreactor is often referred to as "simulated microgravity". Both devices utilize constant reorientation to effectively nullify cumulative sedimentation of particles. Neither, however, can fully reproduce the concurrent lack of structural deformation, displacement of intercellular components and/or reduced mass transfer in the extracellular fluid that occur in actual weightlessness. Parameters including density, viscosity, and even container geometry must each be considered to determine the overall gravity-dependent effects produced by either a clinostat or the RWV bioreactor; in addition, the intended application of these two devices differs considerably. A state of particle "motionlessness" relative to the surrounding bulk fluid, which is nearly analogous to the extracellular environment encountered under weightless conditions, can theoretically be achieved through clinorotation. The RWV bioreactor, on the other hand, while similarly maintaining cells in suspension as they continually "fall" through the medium under 1 g conditions, can also purposefully induce a perfusion of nutrients to and waste from the culture. A clinostat, therefore, is typically used in an attempt to reproduce the quiescent, unstirred fluid conditions achievable on orbit; while the RWV bioreactor ideally creates a low shear, but necessarily mixed, fluid environment that is optimized for suspension culture and tissue growth. Other techniques for exploring altered inertial environments, such as freefall, neutral buoyancy and electromagnetic levitation, can also provide unique insight into how gravity affects biological systems. Ultimately, all underlying biophysical principles thought to give rise to gravity-dependent physiological responses must be identified and thoroughly examined in order to accurately interpret data from flight experiments or ground-based microgravity analogs.

Adipogenesis of human adipose-derived stem cells within three-dimensional hollow fiber-based bioreactors.

PubMed

Gerlach, Jörg C; Lin, Yen-Chih; Brayfield, Candace A; Minteer, Danielle M; Li, Han; Rubin, J Peter; Marra, Kacey G

2012-01-01

To further differentiate adipose-derived stem cells (ASCs) into mature adipocytes and create three-dimensional (3D) adipose tissue in vitro, we applied multicompartment hollow fiber-based bioreactor technology with decentral mass exchange for more physiological substrate gradients and integral oxygenation. We hypothesize that a dynamic 3D perfusion in such a bioreactor will result in longer-term culture of human adipocytes in vitro, thus providing metabolically active tissue serving as a diagnostic model for screening drugs to treat diabetes. ASCs were isolated from discarded human abdominal subcutaneous adipose tissue and then inoculated into dynamic 3D culture bioreactors to undergo adipogenic differentiation. Insulin-stimulated glucose uptake from the medium was assessed with and without TNF-alpha. 3D adipose tissue was generated in the 3D-bioreactors. Immunohistochemical staining indicated that 3D-bioreactor culture displayed multiple mature adipocyte markers with more unilocular morphologies as compared with two-dimensional (2D) cultures. Results of real-time polymerase chain reaction showed 3D-bioreactor treatment had more efficient differentiation in fatty acid-binding protein 4 expression. Repeated insulin stimulation resulted in increased glucose uptake, with a return to baseline between testing. Importantly, TNF-alpha inhibited glucose uptake, an indication of the metabolic activity of the tissue. 3D bioreactors allow more mature adipocyte differentiation of ASCs compared with traditional 2D culture and generate adipose tissue in vitro for up to 2 months. Reproducible metabolic activity of the adipose tissue in the bioreactor was demonstrated, which is potentially useful for drug discovery. We present here, to the best of our knowledge for the first time, the development of a coherent 3D high density fat-like tissue consisting of unilocular structure from primary adipose stem cells in vitro.

Adipogenesis of Human Adipose-Derived Stem Cells Within Three-Dimensional Hollow Fiber-Based Bioreactors

PubMed Central

Gerlach, Jörg C.; Lin, Yen-Chih; Brayfield, Candace A.; Minteer, Danielle M.; Li, Han; Rubin, J. Peter

2012-01-01

To further differentiate adipose-derived stem cells (ASCs) into mature adipocytes and create three-dimensional (3D) adipose tissue in vitro, we applied multicompartment hollow fiber-based bioreactor technology with decentral mass exchange for more physiological substrate gradients and integral oxygenation. We hypothesize that a dynamic 3D perfusion in such a bioreactor will result in longer-term culture of human adipocytes in vitro, thus providing metabolically active tissue serving as a diagnostic model for screening drugs to treat diabetes. ASCs were isolated from discarded human abdominal subcutaneous adipose tissue and then inoculated into dynamic 3D culture bioreactors to undergo adipogenic differentiation. Insulin-stimulated glucose uptake from the medium was assessed with and without TNF-alpha. 3D adipose tissue was generated in the 3D-bioreactors. Immunohistochemical staining indicated that 3D-bioreactor culture displayed multiple mature adipocyte markers with more unilocular morphologies as compared with two-dimensional (2D) cultures. Results of real-time polymerase chain reaction showed 3D-bioreactor treatment had more efficient differentiation in fatty acid-binding protein 4 expression. Repeated insulin stimulation resulted in increased glucose uptake, with a return to baseline between testing. Importantly, TNF-alpha inhibited glucose uptake, an indication of the metabolic activity of the tissue. 3D bioreactors allow more mature adipocyte differentiation of ASCs compared with traditional 2D culture and generate adipose tissue in vitro for up to 2 months. Reproducible metabolic activity of the adipose tissue in the bioreactor was demonstrated, which is potentially useful for drug discovery. We present here, to the best of our knowledge for the first time, the development of a coherent 3D high density fat-like tissue consisting of unilocular structure from primary adipose stem cells in vitro. PMID:21902468

A Multi-Paradigm Modeling Framework to Simulate Dynamic Reciprocity in a Bioreactor

PubMed Central

Kaul, Himanshu; Cui, Zhanfeng; Ventikos, Yiannis

2013-01-01

Despite numerous technology advances, bioreactors are still mostly utilized as functional black-boxes where trial and error eventually leads to the desirable cellular outcome. Investigators have applied various computational approaches to understand the impact the internal dynamics of such devices has on overall cell growth, but such models cannot provide a comprehensive perspective regarding the system dynamics, due to limitations inherent to the underlying approaches. In this study, a novel multi-paradigm modeling platform capable of simulating the dynamic bidirectional relationship between cells and their microenvironment is presented. Designing the modeling platform entailed combining and coupling fully an agent-based modeling platform with a transport phenomena computational modeling framework. To demonstrate capability, the platform was used to study the impact of bioreactor parameters on the overall cell population behavior and vice versa. In order to achieve this, virtual bioreactors were constructed and seeded. The virtual cells, guided by a set of rules involving the simulated mass transport inside the bioreactor, as well as cell-related probabilistic parameters, were capable of displaying an array of behaviors such as proliferation, migration, chemotaxis and apoptosis. In this way the platform was shown to capture not only the impact of bioreactor transport processes on cellular behavior but also the influence that cellular activity wields on that very same local mass transport, thereby influencing overall cell growth. The platform was validated by simulating cellular chemotaxis in a virtual direct visualization chamber and comparing the simulation with its experimental analogue. The results presented in this paper are in agreement with published models of similar flavor. The modeling platform can be used as a concept selection tool to optimize bioreactor design specifications. PMID:23555740

NASA's Space Launch System Advanced Booster Engineering Demonstration and Risk Reduction Efforts

NASA Technical Reports Server (NTRS)

Crumbly, Christopher M.; May, Todd; Dumbacher, Daniel

2012-01-01

The National Aeronautics and Space Administration (NASA) formally initiated the Space Launch System (SLS) development in September 2011, with the approval of the program s acquisition plan, which engages the current workforce and infrastructure to deliver an initial 70 metric ton (t) SLS capability in 2017, while using planned block upgrades to evolve to a full 130 t capability after 2021. A key component of the acquisition plan is a three-phased approach for the first stage boosters. The first phase is to complete the development of the Ares and Space Shuttle heritage 5-segment solid rocket boosters for initial exploration missions in 2017 and 2021. The second phase in the booster acquisition plan is the Advanced Booster Risk Reduction and/or Engineering Demonstration NASA Research Announcement (NRA), which was recently awarded after a full and open competition. The NRA was released to industry on February 9, 2012, and its stated intent was to reduce risks leading to an affordable Advanced Booster and to enable competition. The third and final phase will be a full and open competition for Design, Development, Test, and Evaluation (DDT&E) of the Advanced Boosters. There are no existing boosters that can meet the performance requirements for the 130 t class SLS. The expected thrust class of the Advanced Boosters is potentially double the current 5-segment solid rocket booster capability. These new boosters will enable the flexible path approach to space exploration beyond Earth orbit, opening up vast opportunities including near-Earth asteroids, Lagrange Points, and Mars. This evolved capability offers large volume for science missions and payloads, will be modular and flexible, and will be right-sized for mission requirements. NASA developed the Advanced Booster Engineering Demonstration and/or Risk Reduction NRA to seek industry participation in reducing risks leading to an affordable Advanced Booster that meets the SLS performance requirements. Demonstrations and

Two-liquid-phase bioreactors.

PubMed

Van Sonsbeek, H M; Beeftink, H H; Tramper, J

1993-09-01

The application of two liquid phases that are poorly miscible is a fascinating research topic for biocatalytical conversions because of the promising results. Motives for application include an increase of productivity and achievement of continuous processing, but new limitations arise, e.g., interfacial effects such as biocatalyst accumulation and loss of activity, medium component accumulation, and slow coalescence. Centrifuges, membranes, and immobilization are tools that can overcome part of the problems, but more fundamental knowledge about interfaces and coalescence is still necessary for successful application. For scaleup and further development of processes based on the obtained results, a choice must be made for the configuration of the experimental setup of a bioreactor. Aspects like aeration, shear stress, batch or continuous processing, and immobilization can play an important role. This review article describes these aspects and the proposals that have been made in recent years concerning two-liquid-phase bioreactors. It shows some adaptations to existing bioreactors, such as loop reactors and stirred-tank reactors.

Stirred suspension bioreactors as a novel method to enrich germ cells from pre-pubertal pig testis

PubMed Central

Dores, Camila; Rancourt, Derrick; Dobrinski, Ina

2015-01-01

To study spermatogonial stem cells the heterogeneous testicular cell population first needs to be enriched for undifferentiated spermatogonia, which contain the stem cell population. When working with non-rodent models, this step requires working with large numbers of cells. Available cell separation methods rely on differential properties of testicular cell types such as expression of specific cell surface proteins, size, density or differential adhesion to substrates to separate germ cells from somatic cells. The objective of this study was to develop an approach that allowed germ cell enrichment while providing efficiency of handling large cell numbers. Here we report the use of stirred suspension bioreactors to exploit the adhesion properties of Sertoli cells to enrich cells obtained from pre-pubertal porcine testes for undifferentiated spermatogonia. We also compared the bioreactor approach with an established differential plating method and the combination of both: stirred suspension bioreactor followed by differential plating. After 66 hours of culture, germ cell enrichment in stirred suspension bioreactors provided 7.3±1.0 fold (n=9), differential plating 9.8±2.4 fold (n=6) and combination of both methods resulted in 9.1±0.3 fold enrichment of germ cells from the initial germ cell population (n=3). To document functionality of cells recovered from the bioreactor, we demonstrated that cells retained their functional ability to reassemble seminiferous tubules de novo after grafting to mouse hosts and to support spermatogenesis. These results demonstrate that the stirred suspension bioreactor allows enrichment of germ cells in a controlled and scalable environment providing an efficient method when handling large cell numbers while reducing variability due to handling. PMID:25877677

Role of Bioreactors in Microbial Biomass and Energy Conversion

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Liang; Zhang, Biao; Zhu, Xun

Bioenergy is the world’s largest contributor to the renewable and sustainable energy sector, and it plays a significant role in various energy industries. A large amount of research has contributed to the rapidly evolving field of bioenergy and one of the most important topics is the use of the bioreactor. Bioreactors play a critical role in the successful development of technologies for microbial biomass cultivation and energy conversion. In this chapter, after a brief introduction to bioreactors (basic concepts, configurations, functions, and influencing factors), the applications of the bioreactor in microbial biomass, microbial biofuel conversion, and microbial electrochemical systems aremore » described. Importantly, the role and significance of the bioreactor in the bioenergy process are discussed to provide a better understanding of the use of bioreactors in managing microbial biomass and energy conversion.« less

The stress response system of proteins: Implications for bioreactor scaleup

NASA Technical Reports Server (NTRS)

Goochee, Charles F.

1988-01-01

Animal cells face a variety of environmental stresses in large scale bioreactors, including periodic variations in shear stress and dissolved oxygen concentration. Diagnostic techniques were developed for identifying the particular sources of environmental stresses for animal cells in a given bioreactor configuration. The mechanisms by which cells cope with such stresses was examined. The individual concentrations and synthesis rates of hundreds of intracellular proteins are affected by the extracellular environment (medium composition, dissolved oxygen concentration, ph, and level of surface shear stress). Techniques are currently being developed for quantifying the synthesis rates and concentrations of the intracellular proteins which are most sensitive to environmental stress. Previous research has demonstrated that a particular set of stress response proteins are synthesized by mammalian cells in response to temperature fluctuations, dissolved oxygen deprivation, and glucose deprivation. Recently, it was demonstrated that exposure of human kidney cells to high shear stress results in expression of a completely distinct set of intracellular proteins.

Construction and Characterization of a Novel Vocal Fold Bioreactor

PubMed Central

Zerdoum, Aidan B.; Tong, Zhixiang; Bachman, Brendan; Jia, Xinqiao

2014-01-01

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 µm. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues. PMID:25145349

Construction and characterization of a novel vocal fold bioreactor.

PubMed

Zerdoum, Aidan B; Tong, Zhixiang; Bachman, Brendan; Jia, Xinqiao

2014-08-01

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 µm. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

Development of a Cyclic Strain Bioreactor for Mechanical Enhancement and Assessment of Bioengineered Myocardial Constructs

PubMed Central

Salazar, Betsy H.; Cashion, Avery T.; Dennis, Robert G.; Birla, Ravi K.

2015-01-01

Purpose The purpose of this study was to develop enabling bioreactor technologies using a novel voice coil actuator system for investigating the effects of periodic strain on cardiac patches fabricated with rat cardiomyocytes. Methods The bioengineered muscle constructs used in this study were formed by culturing rat neonatal primary cardiac cells on a fibrin gel. The physical design of the bioreactor was initially conceived using Solidworks to test clearances and perform structural strain analysis. Once the software design phase was completed the bioreactor was assembled using a combination of commercially available, custom machined, and 3-D printed parts. We utilized the bioreactor to evaluate the effect of a 4-hour stretch protocol on the contractile properties of the tissue after which immunohistological assessment of the tissue was also performed. Results An increase in contractile force was observed after the strain protocol of 10% stretch at 1Hz, with no significant increase observed in the control group. Additionally, an increase in cardiac myofibril alignment, connexin 43 expression, and collagen type I distribution were noted. Conclusion In this study we demonstrated the effectiveness of a new bioreactor design to improve contractility of engineered cardiac muscle tissue. PMID:26577484

Development of a Cyclic Strain Bioreactor for Mechanical Enhancement and Assessment of Bioengineered Myocardial Constructs.

PubMed

Salazar, Betsy H; Cashion, Avery T; Dennis, Robert G; Birla, Ravi K

2015-12-01

The purpose of this study was to develop enabling bioreactor technologies using a novel voice coil actuator system for investigating the effects of periodic strain on cardiac patches fabricated with rat cardiomyocytes. The bioengineered muscle constructs used in this study were formed by culturing rat neonatal primary cardiac cells on a fibrin gel. The physical design of the bioreactor was initially conceived using Solidworks to test clearances and perform structural strain analysis. Once the software design phase was completed the bioreactor was assembled using a combination of commercially available, custom machined, and 3-D printed parts. We utilized the bioreactor to evaluate the effect of a 4-h stretch protocol on the contractile properties of the tissue after which immunohistological assessment of the tissue was also performed. An increase in contractile force was observed after the strain protocol of 10% stretch at 1 Hz, with no significant increase observed in the control group. Additionally, an increase in cardiac myofibril alignment, connexin 43 expression, and collagen type I distribution were noted. In this study we demonstrated the effectiveness of a new bioreactor design to improve contractility of engineered cardiac muscle tissue.

Development of an energy-saving anaerobic hybrid membrane bioreactors for 2-chlorophenol-contained wastewater treatment.

PubMed

Wang, Yun-Kun; Pan, Xin-Rong; Sheng, Guo-Ping; Li, Wen-Wei; Shi, Bing-Jing; Yu, Han-Qing

2015-12-01

A novel energy-saving anaerobic hybrid membrane bioreactor (AnHMBR) with mesh filter, which takes advantage of anaerobic membrane bioreactor and fixed-bed biofilm reactor, is developed for low-strength 2-chlorophenol (2-CP)-contained wastewater treatment. In this system, the anaerobic membrane bioreactor is stuffed with granular activated carbon to construct an anaerobic hybrid fixed-bed biofilm membrane bioreactor. The effluent turbidity from the AnHMBR system was low during most of the operation period, and the chemical oxygen demand and 2-CP removal efficiencies averaged 82.3% and 92.6%, respectively. Furthermore, a low membrane fouling rate was achieved during the operation. During the AnHMBR operation, the only energy consumption was for feed pump. And a low energy demand of 0.0045-0.0063kWhm(-3) was estimated under the current operation conditions. All these results demonstrated that this novel AnHMBR is a sustainable technology for treating 2-CP-contained wastewater. Copyright © 2014 Elsevier Ltd. All rights reserved.

In Situ Bioreactor

ScienceCinema

Blackwelder, Brad

2018-05-11

At Idaho National Laboratory, researchers have developed bioreactor technology that permits identification, bioremediation testing and treatment at the source using naturally occurring microbes to disarm contaminants.

Bioreactor Technology in Cardiovascular Tissue Engineering

NASA Astrophysics Data System (ADS)

Mertsching, H.; Hansmann, J.

Cardiovascular tissue engineering is a fast evolving field of biomedical science and technology to manufacture viable blood vessels, heart valves, myocar-dial substitutes and vascularised complex tissues. In consideration of the specific role of the haemodynamics of human circulation, bioreactors are a fundamental of this field. The development of perfusion bioreactor technology is a consequence of successes in extracorporeal circulation techniques, to provide an in vitro environment mimicking in vivo conditions. The bioreactor system should enable an automatic hydrodynamic regime control. Furthermore, the systematic studies regarding the cellular responses to various mechanical and biochemical cues guarantee the viability, bio-monitoring, testing, storage and transportation of the growing tissue.

Performance of an anaerobic, static bed, fixed film bioreactor for chlorinated solvent treatment

USGS Publications Warehouse

Lorah, Michelle M.; Walker, Charles; Graves, Duane

2015-01-01

Anaerobic, fixed film, bioreactors bioaugmented with a dechlorinating microbial consortium were evaluated as a potential technology for cost effective, sustainable, and reliable treatment of mixed chlorinated ethanes and ethenes in groundwater from a large groundwater recovery system. Bench- and pilot-scale testing at about 3 and 13,500 L, respectively, demonstrated that total chlorinated solvent removal to less than the permitted discharge limit of 100 μg/L. Various planned and unexpected upsets, interruptions, and changes demonstrated the robustness and reliability of the bioreactor system, which handled the operational variations with no observable change in performance. Key operating parameters included an adequately long hydraulic retention time for the surface area, a constant supply of electron donor, pH control with a buffer to minimize pH variance, an oxidation reduction potential of approximately −200 millivolts or lower, and a well-adapted biomass capable of degrading the full suite of chlorinated solvents in the groundwater. Results indicated that the current discharge criteria can be met using a bioreactor technology that is less complex and has less downtime than the sorption based technology currently being used to treat the groundwater.

Effect of sudden addition of PCE and bioreactor coupling to ZVI filters on performance of fluidized bed bioreactors operated in simultaneous electron acceptor modes.

PubMed

Moreno-Medina, C U; Poggi-Varaldo, Hector M; Breton-Deval, L; Rinderknecht-Seijas, N

2017-11-01

The present work evaluated the effects of (i) feeding a water contaminated with 80 mg/L PCE to bioreactors seeded with inoculum not acclimated to PCE, (ii) coupling ZVI side filters to bioreactors, and (iii) working in different biological regimes, i.e., simultaneous methanogenic aeration and simultaneous methanogenic-denitrifying regimes, on fluidized bed bioreactor performance. Simultaneous electron acceptors refer to the simultaneous presence of two compounds operating as final electron acceptors in the biological respiratory chain (e.g., use of either O 2 or NO 3 - in combination with a methanogenic environment) in a bioreactor or environmental niche. Four lab-scale, mesophilic, fluidized bed bioreactors (bioreactors) were implemented. Two bioreactors were operated as simultaneous methanogenic-denitrifying (MD) units, whereas the other two were operated in partially aerated methanogenic (PAM) mode. In the first period, all bioreactors received a wastewater with 1 g chemical oxygen demand of methanol per liter (COD-methanol/L). In a second period, all the bioreactors received the wastewater plus 80 mg perchloroethylene (PCE)/L; at the start of period 2, one MD and one PAM were coupled to side sand-zero valent iron filters (ZVI). All bioreactors were inoculated with a microbial consortium not acclimated to PCE. In this work, the performance of the full period 1 and the first 60 days of period 2 is reported and discussed. The COD removal efficiency and the nitrate removal efficiency of the bioreactors essentially did not change between period 1 and period 2, i.e., upon PCE addition. On the contrary, specific methanogenic activity in PAM bioreactors (both with and without coupled ZVI filter) significantly decreased. This was consistent with a sharp fall of methane productivity in those bioreactors in period 2. During period 2, PCE removals in the range 86 to 97 % were generally observed; the highest removal corresponded to PAM bioreactors along with the

Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes

PubMed Central

2011-01-01

Background Embryonic stem cells (ESCs) can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs). However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Results Murine D3-MHC-neor ESCs formed embyroid bodies (EBs) and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin) was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. Conclusions This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC differentiation protocols within

Validation of Solar Sail Simulations for the NASA Solar Sail Demonstration Project

NASA Technical Reports Server (NTRS)

Braafladt, Alexander C.; Artusio-Glimpse, Alexandra B.; Heaton, Andrew F.

2014-01-01

NASA's Solar Sail Demonstration project partner L'Garde is currently assembling a flight-like sail assembly for a series of ground demonstration tests beginning in 2015. For future missions of this sail that might validate solar sail technology, it is necessary to have an accurate sail thrust model. One of the primary requirements of a proposed potential technology validation mission will be to demonstrate solar sail thrust over a set time period, which for this project is nominally 30 days. This requirement would be met by comparing a L'Garde-developed trajectory simulation to the as-flown trajectory. The current sail simulation baseline for L'Garde is a Systems Tool Kit (STK) plug-in that includes a custom-designed model of the L'Garde sail. The STK simulation has been verified for a flat plate model by comparing it to the NASA-developed Solar Sail Spaceflight Simulation Software (S5). S5 matched STK with a high degree of accuracy and the results of the validation indicate that the L'Garde STK model is accurate enough to meet the potential future mission requirements. Additionally, since the L'Garde sail deviates considerably from a flat plate, a force model for a non-flat sail provided by L'Garde sail was also tested and compared to a flat plate model in S5. This result will be used in the future as a basis of comparison to the non-flat sail model being developed for STK.

Requirements and feasibility study of flight demonstration of Active Controls Technology (ACT) on the NASA 515 airplane

NASA Technical Reports Server (NTRS)

Gordon, C. K.

1975-01-01

A preliminary design study was conducted to evaluate the suitability of the NASA 515 airplane as a flight demonstration vehicle, and to develop plans, schedules, and budget costs for fly-by-wire/active controls technology flight validation in the NASA 515 airplane. The preliminary design and planning were accomplished for two phases of flight validation.

Design of well and groove microchannel bioreactors for cell culture.

PubMed

Korin, Natanel; Bransky, Avishay; Khoury, Maria; Dinnar, Uri; Levenberg, Shulamit

2009-03-01

microm depth) and well bioreactors (35 microm depth) was experimentally examined at various flow rates of medium perfusion and compared to cell culture in regular flat microchannels. It was shown that the wells and the grooves enable a one order of magnitude increase in the maximum perfusion rate compared to planar microchannels. Altogether, the study demonstrates that the proper design and use of microgroove/well bioreactors may be highly beneficial for cell culture assays.

A Review of the NASA MLAS Flight Demonstration

NASA Technical Reports Server (NTRS)

Taylor, Anthony P.; Kelley, Christopher; Manger, Eldred; Peterson, David; Hahn, Jeffrey; Yuchnovicz, Daniel

2011-01-01

The NASA Engineering and Safety Center (NESC) has tested the Max Launch Abort System (MLAS) as a risk-mitigation design should problems arise with the baseline Orion spacecraft launch abort design. The Max in MLAS is not Maximum, but rather dedicated to Max Faget, the renowned NASA Spacecraft designer. In July 2009, the mission was flown, with great success, from the NASA Wallops Flight Facility. The MLAS flight test vehicle prototype consists of a boost skirt, coast skirt, and the MLAS fairing itself, which houses an Orion Command Module (CM) boilerplate. The objective of the MLAS flight test is to reorient the fairing with the CM, weighing approximately 29,000 lbs and traveling 290 fps, 180 degrees to an orientation suitable for the release of the CM during a pad abort or low altitude abort. The boost and coast skirts provide the necessary thrust and stability to establish the flight test conditions and are released prior to the reorientation of the fairing. A secondary test objective after successful release of the CM from the fairing is to demonstrate the removal of the CM forward bay cover (FBC) with the CM drogue parachutes, and subsequent deployment of the CM main parachutes attached to the FBC. Although multiple parachute deployments are used in the MLAS flight test vehicle to complete its objective, there are only two parachute types employed in the flight test. Five of the nine parachutes used for MLAS are 27.6 ft DO ribbon parachutes already proven as a spin/stall parachute for military aircraft, and the remaining four are G-12 cargo parachutes modified for increased strength and reefing. This paper presents an overview of the 27.6 ft DO ribbon parachute system employed on the MLAS flight test vehicle for coast skirt separation, fairing reorientation, and as CM drogue parachutes. Discussion will include: the process used to select this design; descriptions of all components of the parachute system; the minor modifications necessary to adapt the parachute

The Effect of Simulated Microgravity Environment of RWV Bioreactors on Surface Reactions and Adsorption of Serum Proteins on Bone-bioactive Microcarriers

NASA Technical Reports Server (NTRS)

Radin, Shula; Ducheyne, P.; Ayyaswamy, P. S.

2003-01-01

Biomimetically modified bioactive materials with bone-like surface properties are attractive candidates for use as microcarriers for 3-D bone-like tissue engineering under simulated microgravity conditions of NASA designed rotating wall vessel (RWV) bioreactors. The simulated microgravity environment is attainable under suitable parametric conditions of the RWV bioreactors. Ca-P containing bioactive glass (BG), whose stimulatory effect on bone cell function had been previously demonstrated, was used in the present study. BG surface modification via reactions in solution, resulting formation of bone-like minerals at the surface and adsorption of serum proteins is critical for obtaining the stimulatory effect. In this paper, we report on the major effects of simulated microgravity conditions of the RWV on the BG reactions surface reactions and protein adsorption in physiological solutions. Control tests at normal gravity were conducted at static and dynamic conditions. The study revealed that simulated microgravity remarkably enhanced reactions involved in the BG surface modification, including BG dissolution, formation of bone-like minerals at the surface and adsorption of serum proteins. Simultaneously, numerical models were developed to simulate the mass transport of chemical species to and from the BG surface under normal gravity and simulated microgravity conditions. The numerical results showed an excellent agreement with the experimental data at both testing conditions.

Comparison between a moving bed membrane bioreactor and a conventional membrane bioreactor on organic carbon and nitrogen removal.

PubMed

Yang, Shuai; Yang, Fenglin; Fu, Zhimin; Lei, Ruibo

2009-04-01

A membrane bioreactor filled with carriers instead of activated sludge named a moving bed membrane bioreactor (MBMBR) was investigated for simultaneously removing organic carbon and nitrogen in wastewater. Its performance was compared with a conventional membrane bioreactor (CMBR) at various influent COD/TN ratios of 8.9-22.1. The operational parameters were optimized to increase the treatment efficiency. COD removal efficiency averaged at 95.6% and 96.2%, respectively, for MBMBR and CMBR during the 4 months experimental period. The MBMBR system demonstrated good performance on nitrogen removal at different COD/TN ratios. When COD/TN was 8.9 and the total nitrogen (TN) load was 7.58 mg/l h, the TN and ammonium nitrogen removal efficiencies of the MBMBR were maintained over 70.0% and 80.0%, respectively, and the removed total nitrogen (TN) load reached to 5.31 mg/l h. Multifunctional microbial reactions in the carrier, such as simultaneous nitrification and denitrification (SND), play important roles in nitrogen removal. In comparison, the CMBR did not perform so well. Its TN removal was not stable, and the removed total nitrogen (TN) load was only 1.02 mg/l h at COD/TN ratio 8.9. The specific oxygen utilization rate (SOUR) showed that the biofilm has a better microbial activity than an activated sludge. Nevertheless, the membrane fouling behavior was more severe in the MBMBR than in the CMBR due to a thick and dense cake layer formed on the membrane surface, which was speculated to be caused by the filamentous bacteria in the MBMBR.

Denitrifying bioreactor clogging potential during wastewater treatment.

PubMed

Christianson, Laura E; Lepine, Christine; Sharrer, Kata L; Summerfelt, Steven T

2016-11-15

Chemoheterotrophic denitrification technologies using woodchips as a solid carbon source (i.e., woodchip bioreactors) have been widely trialed for treatment of diffuse-source agricultural nitrogen pollution. There is growing interest in the use of this simple, relatively low-cost biological wastewater treatment option in waters with relatively higher total suspended solids (TSS) and chemical oxygen demand (COD) such as aquaculture wastewater. This work: (1) evaluated hydraulic retention time (HRT) impacts on COD/TSS removal, and (2) assessed the potential for woodchip clogging under this wastewater chemistry. Four pilot-scale woodchip denitrification bioreactors operated for 267 d showed excellent TSS removal (>90%) which occurred primarily near the inlet, and that COD removal was maximized at lower HRTs (e.g., 56% removal efficiency and 25 g of COD removed per m 3 of bioreactor per d at a 24 h HRT). However, influent wastewater took progressively longer to move into the woodchips likely due to a combination of (1) woodchip settling, (2) clogging due to removed wastewater solids and/or accumulated bacterial growth, and (3) the pulsed flow system pushing the chips away from the inlet. The bioreactor that received the highest loading rate experienced the most altered hydraulics. Statistically significant increases in woodchip P content over time in woodchip bags placed near the bioreactor outlets (0.03 vs 0.10%P 2 O 5 ) and along the bioreactor floor (0.04 vs. 0.12%P 2 O 5 ) confirmed wastewater solids were being removed and may pose a concern for subsequent nutrient mineralization and release. Nevertheless, the excellent nitrate-nitrogen and TSS removal along with notable COD removal indicated woodchip bioreactors are a viable water treatment technology for these types of wastewaters given they are used downstream of a filtration device. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Development of Fundamental Technologies for Micro Bioreactors

NASA Astrophysics Data System (ADS)

Sato, Kiichi; Kitamori, Takehiko

This chapter reviews the development of fundamental technologies required for microchip-based bioreactors utilizing living mammalian cells and pressure driven flow. The most important factor in the bioreactor is the cell culture. For proper cell culturing, continuous medium supply from a microfluidic channel and appropriate modification of the channel surface to accommodate cell attachment is required. Moreover, the medium flow rate should be chosen carefully, because shear stress affects cell activity. The techniques presented here could be applied to the development of micro bioreactors such as microlivers, pigment production by plant cells, and artificial insemination.

Space Launch System NASA Research Announcement Advanced Booster Engineering Demonstration and/or Risk Reduction

NASA Technical Reports Server (NTRS)

Crumbly, Christopher M.; Craig, Kellie D.

2011-01-01

The intent of the Advanced Booster Engineering Demonstration and/or Risk Reduction (ABEDRR) effort is to: (1) Reduce risks leading to an affordable Advanced Booster that meets the evolved capabilities of SLS (2) Enable competition by mitigating targeted Advanced Booster risks to enhance SLS affordability. Key Concepts (1) Offerors must propose an Advanced Booster concept that meets SLS Program requirements (2) Engineering Demonstration and/or Risk Reduction must relate to the Offeror s Advanced Booster concept (3) NASA Research Announcement (NRA) will not be prescriptive in defining Engineering Demonstration and/or Risk Reduction

Biodegradation of high concentrations of benzene vapors in a two phase partition stirred tank bioreactor.

PubMed

Karimi, Ali; Golbabaei, Farideh; Neghab, Masoud; Pourmand, Mohammad Reza; Nikpey, Ahmad; Mohammad, Kazem; Mehrnia, Momammad Reza

2013-01-15

The present study examined the biodegradation rate of benzene vapors in a two phase stirred tank bioreactor by a bacterial consortium obtained from wastewater of an oil industry refinery house. Initially, the ability of the microbial consortium for degrading benzene was evaluated before running the bioreactor. The gaseous samples from inlet and outlet of bioreactor were directly injected into a gas chromatograph to determine benzene concentrations. Carbone oxide concentration at the inlet and outlet of bioreactor were also measured with a CO2 meter to determine the mineralization rate of benzene. Influence of the second non-aqueous phase (silicon oil) has been emphasized, so at the first stage the removal efficiency (RE) and elimination capacity (EC) of benzene vapors were evaluated without any organic phase and in the second stage, 10% of silicon oil was added to bioreactor media as an organic phase. Addition of silicon oil increased the biodegradation performance up to an inlet loading of 5580 mg/m3, a condition at which, the elimination capacity and removal efficiency were 181 g/m3/h and 95% respectively. The elimination rate of benzene increased by 38% in the presence of 10% of silicone oil. The finding of this study demonstrated that two phase partition bioreactors (TPPBs) are potentially effective tools for the treatment of gas streams contaminated with high concentrations of poorly water soluble organic contaminant, such as benzene.

BIOREACTOR LANDFILLS, THEORETICAL ADVANTAGES AND RESEARCH CHALLENGES

EPA Science Inventory

Bioreactor landfills are municipal solid waste landfills that utilize bulk liquids in an effort to accelerate solid waste degradation. There are few potential benefits for operating a MSW landfill as a bioreactor. These include leachate treatment and management, increase in the s...

Membrane Bioreactor With Pressure Cycle

NASA Technical Reports Server (NTRS)

Efthymiou, George S.; Shuler, Michael L.

1991-01-01

Improved class of multilayer membrane bioreactors uses convention forced by differences in pressure to overcome some of diffusional limitations of prior bioreactors. In reactor of new class, flow of nutrient solution reduces adverse gradients of concentration, keeps cells supplied with fresh nutrient, and sweeps away products faster than diffusion alone. As result, overall yield and rate of reaction increased. Pressures in sweeping gas and nutrient alternated to force nutrient liquid into and out of biocatalyst layer through hyrophilic membrane.

Bioreactors Drive Advances in Tissue Engineering

NASA Technical Reports Server (NTRS)

2012-01-01

It was an unlikely moment for inspiration. Engineers David Wolf and Ray Schwarz stopped by their lab around midday. Wolf, of Johnson Space Center, and Schwarz, with NASA contractor Krug Life Sciences (now Wyle Laboratories Inc.), were part of a team tasked with developing a unique technology with the potential to enhance medical research. But that wasn t the focus at the moment: The pair was rounding up colleagues interested in grabbing some lunch. One of the lab s other Krug engineers, Tinh Trinh, was doing something that made Wolf forget about food. Trinh was toying with an electric drill. He had stuck the barrel of a syringe on the bit; it spun with a high-pitched whirr when he squeezed the drill s trigger. At the time, a multidisciplinary team of engineers and biologists including Wolf, Schwarz, Trinh, and project manager Charles D. Anderson, who formerly led the recovery of the Apollo capsules after splashdown and now worked for Krug was pursuing the development of a technology called a bioreactor, a cylindrical device used to culture human cells. The team s immediate goal was to grow human kidney cells to produce erythropoietin, a hormone that regulates red blood cell production and can be used to treat anemia. But there was a major barrier to the technology s success: Moving the liquid growth media to keep it from stagnating resulted in turbulent conditions that damaged the delicate cells, causing them to quickly die. The team was looking forward to testing the bioreactor in space, hoping the device would perform more effectively in microgravity. But on January 28, 1986, the Space Shuttle Challenger broke apart shortly after launch, killing its seven crewmembers. The subsequent grounding of the shuttle fleet had left researchers with no access to space, and thus no way to study the effects of microgravity on human cells. As Wolf looked from Trinh s syringe-capped drill to where the bioreactor sat on a workbench, he suddenly saw a possible solution to both

Hydrofocusing Bioreactor for Three-Dimensional Cell Culture

NASA Technical Reports Server (NTRS)

Gonda, Steve R.; Spaulding, Glenn F.; Tsao, Yow-Min D.; Flechsig, Scott; Jones, Leslie; Soehnge, Holly

2003-01-01

The hydrodynamic focusing bioreactor (HFB) is a bioreactor system designed for three-dimensional cell culture and tissue-engineering investigations on orbiting spacecraft and in laboratories on Earth. The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear culture environment simultaneously with the "herding" of suspended cells, tissue assemblies, and air bubbles. Under development for use in the Biotechnology Facility on the International Space Station, the HFB has successfully grown large three-dimensional, tissuelike assemblies from anchorage-dependent cells and grown suspension hybridoma cells to high densities. The HFB, based on the principle of hydrodynamic focusing, provides the capability to control the movement of air bubbles and removes them from the bioreactor without degrading the low-shear culture environment or the suspended three-dimensional tissue assemblies. The HFB also provides unparalleled control over the locations of cells and tissues within its bioreactor vessel during operation and sampling.

A Dual-Mode Bioreactor System for Tissue Engineered Vascular Models.

PubMed

Bono, N; Meghezi, S; Soncini, M; Piola, M; Mantovani, D; Fiore, Gianfranco Beniamino

2017-06-01

In the past decades, vascular tissue engineering has made great strides towards bringing engineered vascular tissues to the clinics and, in parallel, obtaining in-lab tools for basic research. Herein, we propose the design of a novel dual-mode bioreactor, useful for the fabrication (construct mode) and in vitro stimulation (culture mode) of collagen-based tubular constructs. Collagen-based gels laden with smooth muscle cells (SMCs) were molded directly within the bioreactor culture chamber. Based on a systematic characterization of the bioreactor culture mode, constructs were subjected to 10% cyclic strain at 0.5 Hz for 5 days. The effects of cyclic stimulation on matrix re-arrangement and biomechanical/viscoelastic properties were examined and compared vs. statically cultured constructs. A thorough comparison of cell response in terms of cell localization and expression of contractile phenotypic markers was carried out as well. We found that cyclic stimulation promoted cell-driven collagen matrix bi-axial compaction, enhancing the mechanical strength of strained samples with respect to static controls. Moreover, cyclic strain positively affected SMC behavior: cells maintained their contractile phenotype and spread uniformly throughout the whole wall thickness. Conversely, static culture induced a noticeable polarization of cell distribution to the outer rim of the constructs and a sharp reduction in total cell density. Overall, coupling the use of a novel dual-mode bioreactor with engineered collagen-gel-based tubular constructs demonstrated to be an interesting technology to investigate the modulation of cell and tissue behavior under controlled mechanically conditioned in vitro maturation.

NASA's Space Launch System Advanced Booster Engineering Demonstration and/or Risk Reduction Efforts

NASA Technical Reports Server (NTRS)

Crumbly, Christopher M.; Dumbacher, Daniel L.; May, Todd A.

2012-01-01

The National Aeronautics and Space Administration (NASA) formally initiated the Space Launch System (SLS) development in September 2011, with the approval of the program s acquisition plan, which engages the current workforce and infrastructure to deliver an initial 70 metric ton (t) SLS capability in 2017, while using planned block upgrades to evolve to a full 130 t capability after 2021. A key component of the acquisition plan is a three-phased approach for the first stage boosters. The first phase is to complete the development of the Ares and Space Shuttle heritage 5-segment solid rocket boosters (SRBs) for initial exploration missions in 2017 and 2021. The second phase in the booster acquisition plan is the Advanced Booster Risk Reduction and/or Engineering Demonstration NASA Research Announcement (NRA), which was recently awarded after a full and open competition. The NRA was released to industry on February 9, 2012, with a stated intent to reduce risks leading to an affordable advanced booster and to enable competition. The third and final phase will be a full and open competition for Design, Development, Test, and Evaluation (DDT&E) of the advanced boosters. There are no existing boosters that can meet the performance requirements for the 130 t class SLS. The expected thrust class of the advanced boosters is potentially double the current 5-segment solid rocket booster capability. These new boosters will enable the flexible path approach to space exploration beyond Earth orbit (BEO), opening up vast opportunities including near-Earth asteroids, Lagrange Points, and Mars. This evolved capability offers large volume for science missions and payloads, will be modular and flexible, and will be right-sized for mission requirements. NASA developed the Advanced Booster Engineering Demonstration and/or Risk Reduction NRA to seek industry participation in reducing risks leading to an affordable advanced booster that meets the SLS performance requirements

BIOREACTOR LANDFILL DESIGN

EPA Science Inventory

Modern landfill design entails many elements including foundations, liner systems, leachate collection systems, stormwater control systems, slope stability considerations, leachate management systems, gas extraction systems, and capping and closure. The use of bioreactor technolo...

A Long-Term Cultivation of an Anaerobic Methane-Oxidizing Microbial Community from Deep-Sea Methane-Seep Sediment Using a Continuous-Flow Bioreactor

PubMed Central

Aoki, Masataka; Ehara, Masayuki; Saito, Yumi; Yoshioka, Hideyoshi; Miyazaki, Masayuki; Saito, Yayoi; Miyashita, Ai; Kawakami, Shuji; Yamaguchi, Takashi; Ohashi, Akiyoshi; Nunoura, Takuro; Takai, Ken; Imachi, Hiroyuki

2014-01-01

Anaerobic oxidation of methane (AOM) in marine sediments is an important global methane sink, but the physiological characteristics of AOM-associated microorganisms remain poorly understood. Here we report the cultivation of an AOM microbial community from deep-sea methane-seep sediment using a continuous-flow bioreactor with polyurethane sponges, called the down-flow hanging sponge (DHS) bioreactor. We anaerobically incubated deep-sea methane-seep sediment collected from the Nankai Trough, Japan, for 2,013 days in the bioreactor at 10°C. Following incubation, an active AOM activity was confirmed by a tracer experiment using 13C-labeled methane. Phylogenetic analyses demonstrated that phylogenetically diverse Archaea and Bacteria grew in the bioreactor. After 2,013 days of incubation, the predominant archaeal components were anaerobic methanotroph (ANME)-2a, Deep-Sea Archaeal Group, and Marine Benthic Group-D, and Gammaproteobacteria was the dominant bacterial lineage. Fluorescence in situ hybridization analysis showed that ANME-1 and -2a, and most ANME-2c cells occurred without close physical interaction with potential bacterial partners. Our data demonstrate that the DHS bioreactor system is a useful system for cultivating fastidious methane-seep-associated sedimentary microorganisms. PMID:25141130

An automated perfusion bioreactor for the streamlined production of engineered osteogenic grafts.

PubMed

Ding, Ming; Henriksen, Susan S; Wendt, David; Overgaard, Søren

2016-04-01

A computer-controlled perfusion bioreactor was developed for the streamlined production of engineered osteogenic grafts. This system automated the required bioprocesses, from the initial filling of the system through the phases of cell seeding and prolonged cell/tissue culture. Flow through chemo-optic micro-sensors allowed to non-invasively monitor the levels of oxygen and pH in the perfused culture medium throughout the culture period. To validate its performance, freshly isolated ovine bone marrow stromal cells were directly seeded on porous scaffold granules (hydroxyapatite/β-tricalcium-phosphate/poly-lactic acid), bypassing the phase of monolayer cell expansion in flasks. Either 10 or 20 days after culture, engineered cell-granule grafts were implanted in an ectopic mouse model to quantify new bone formation. After four weeks of implantation, histomorphometry showed more bone in bioreactor-generated grafts than cell-free granule controls, while bone formation did not show significant differences between 10 days and 20 days of incubation. The implanted granules without cells had no bone formation. This novel perfusion bioreactor has revealed the capability of activation larger viable bone graft material, even after shorter incubation time of graft material. This study has demonstrated the feasibility of engineering osteogenic grafts in an automated bioreactor system, laying the foundation for a safe, regulatory-compliant, and cost-effective manufacturing process. © 2015 Wiley Periodicals, Inc.

Production of oncolytic adenovirus and human mesenchymal stem cells in a single-use, Vertical-Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier-based cell culture processes.

PubMed

Sousa, Marcos F Q; Silva, Marta M; Giroux, Daniel; Hashimura, Yas; Wesselschmidt, Robin; Lee, Brian; Roldão, António; Carrondo, Manuel J T; Alves, Paula M; Serra, Margarida

2015-01-01

successfully carried out for two different microcarrier-based cell cultures. Ultimately, the data herein generated demonstrate the potential of Vertical-Wheel bioreactors as a new scalable biomanufacturing platform for microcarrier-based cell cultures of complex biopharmaceuticals. © 2015 American Institute of Chemical Engineers.

Bacterial community dynamics during start-up of a trickle-bed bioreactor degrading aromatic compounds.

PubMed

Stoffels, M; Amann, R; Ludwig, W; Hekmat, D; Schleifer, K H

1998-03-01

This study was performed with a laboratory-scale fixed-bed bioreactor degrading a mixture of aromatic compounds (Solvesso100). The starter culture for the bioreactor was prepared in a fermentor with a wastewater sample of a care painting facility as the inoculum and Solvesso100 as the sole carbon source. The bacterial community dynamics in the fermentor and the bioreactor were examined by a conventional isolation procedure and in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotides. Two significant shifts in the bacterial community structure could be demonstrated. The original inoculum from the wastewater of the car factory was rich in proteobacteria of the alpha and beta subclasses, while the final fermentor enrichment was dominated by bacteria closely related to Pseudomonas putida or Pseudomonas mendocina, which both belong to the gamma subclass of the class Proteobacteria. A second significant shift was observed when the fermentor culture was transferred as inoculum to the trickle-bed bioreactor. The community structure in the bioreactor gradually returned to a higher complexity, with the dominance of beta and alpha subclass proteobacteria, whereas the gamma subclass proteobacteria sharply declined. Obviously, the preceded pollutant adaptant did not lead to a significant enrichment of bacteria that finally dominated in the trickle-bed bioreactor. In the course of experiments, three new 16S as well as 23S rRNA-targeted probes for beta subclass proteobacteria were designed, probe SUBU1237 for the genera Burkholderia and Sutterella, probe ALBO34a for the genera Alcaligenes and Bordetella, and probe Bcv13b for Burkholderia cepacia and Burkholderia vietnamiensis. Bacteria hybridizing with the probe Bcv13b represented the main Solvesso100-degrading population in the reactor.

NASA Radar Demonstrates Ability to Foresee Sinkholes

NASA Image and Video Library

2014-03-06

Analyses by NASA UAVSAR radar performed after the Bayou Corne, La., sinkhole formed, show it was able to detect precursory ground surface movement of up to 10.2 inches 260 millimeters more than a month before the sinkhole collapsed in Aug. 2012.

A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids

PubMed Central

Madeddu, Denise; Cerino, Giulia; Falco, Angela; Frati, Caterina; Gallo, Diego; Deriu, Marco A.; Falvo D’Urso Labate, Giuseppe; Quaini, Federico; Audenino, Alberto; Morbiducci, Umberto

2016-01-01

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future. PMID:27144306

A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids.

PubMed

Massai, Diana; Isu, Giuseppe; Madeddu, Denise; Cerino, Giulia; Falco, Angela; Frati, Caterina; Gallo, Diego; Deriu, Marco A; Falvo D'Urso Labate, Giuseppe; Quaini, Federico; Audenino, Alberto; Morbiducci, Umberto

2016-01-01

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future.

Bioreactor engineering using disposable technology for enhanced production of hCTLA4Ig in transgenic rice cell cultures.

PubMed

Kwon, Jun-Young; Yang, Yong-Suk; Cheon, Su-Hwan; Nam, Hyung-Jin; Jin, Gi-Hong; Kim, Dong-Il

2013-09-01

Two kinds of disposable bioreactors, air-lift disposable bioreactors (ADB) and wave disposable bioreactors (WDB) were compared with stirred-tank reactors (5-L STR). These bioreactors were successfully applied to transgenic rice cell cultures for the production of recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). In both systems, a fed-batch culture method was used to produce hCTLA4Ig efficiently by feeding concentrated amino acids and production levels were enhanced when dissolved oxygen (DO) level was regulated at 30% using pure oxygen sparging. Agitation and aeration rate during cultivation in ADB and WDB were determined by the same mixing time. The results in both disposable bioreactors showed similar values in maximum cell density (11.9 gDCW/L and 12.6 gDCW/L), doubling time (4.8- and 5.0-day), and maximum hCTLA4Ig concentration (43.7 and 43.3 mg/L). Relatively higher cell viability was sustained in the ADB whereas hCTLA4Ig productivity was 1.2-fold higher than that in WDB. The productivity was improved by increasing aeration rate (0.2 vvm). Overall, our experiments demonstrate pneumatically driven disposable bioreactors are applicable for the production of recombinant proteins in plant cell cultures. These results will be useful for development and scale-up studies of disposable bioreactor systems for transgenic plant cell cultures. Copyright © 2013 Wiley Periodicals, Inc.

Bioreactors for plant cells: hardware configuration and internal environment optimization as tools for wider commercialization.

PubMed

Georgiev, Milen I; Weber, Jost

2014-07-01

Mass production of value-added molecules (including native and heterologous therapeutic proteins and enzymes) by plant cell culture has been demonstrated as an efficient alternative to classical technologies [i.e. natural harvest and chemical (semi)synthesis]. Numerous proof-of-concept studies have demonstrated the feasibility of scaling up plant cell culture-based processes (most notably to produce paclitaxel) and several commercial processes have been established so far. The choice of a suitable bioreactor design (or modification of an existing commercially available reactor) and the optimization of its internal environment have been proven as powerful tools toward successful mass production of desired molecules. This review highlights recent progress (mostly in the last 5 years) in hardware configuration and optimization of bioreactor culture conditions for suspended plant cells.

Lithium-ion Battery Demonstration for the 2007 NASA Desert Research and Technology Studies (Desert RATS) Program

NASA Technical Reports Server (NTRS)

Bennett, William; Baldwin, Richard

2007-01-01

The NASA Glenn Research Center (GRC) Electrochemistry Branch designed and produced five lithium-ion battery packs for demonstration in a portable life support system (PLSS) on spacesuit simulators. The experimental batteries incorporated advanced, NASA-developed electrolytes and included internal protection against over-current, over-discharge and over-temperature. The 500-gram batteries were designed to deliver a constant power of 38 watts over 103 minutes of discharge time (130 Wh/kg). Battery design details are described and field and laboratory test results are summarized.

Spaceflight bioreactor studies of cells and tissues.

PubMed

Freed, Lisa E; Vunjak-Novakovic, Gordana

2002-01-01

well-being (loss of muscle and skeletal tissues [15-17]) and gene- and cell-level responses to the mechanical environment [13,14,18]. All five of the spaceflight bioreactor studies described above utilized three-dimensional cell culture systems in which the cells were associated with biodegradable polymer scaffolds [17], collagen gel [16], or microcarrier beads [13-15,18] in order to promote the expression of differentiated cell function. In four of the five spaceflight bioreactor studies [15-18], cells were cultured in perfused vessels (cartridges or rotating bioreactors) within recirculating loops designed to maintain medium composition within target ranges by a combination of gas exchange and fresh medium supply. Future spaceflight studies of cells and tissues are likely to involve a three-dimensional culture system, to promote cellular differentiation, and perfusion with or without rotation, to provide a gravity-independent mechanism for fluid mixing and mass transport. Previous spaceflight studies have guided the ongoing development of NASA flight hardware for the ISS (e.g. the EDU-2 and the CCU). This next generation of hardware will have extended operational capabilities including on-line microscopy, in-line sensors for the monitoring and control of metabolic parameters, modular design for replicate cultures, and, perhaps most importantly of all, compatibility with the ISS centrifuge. The latter will permit in-flight, 1 g control cultures, and thereby allow the experimental variable to be gravity itself rather than the more general "spaceflight environment". Technical limitations of spaceflight studies (e.g. allowable size, mass, and power) continue to motivate a creative approach to system design and to result in "spin-off" technologies (e.g. the STLV) for ground-based cell and tissue culture research. The increasing scientific and medical relevance of this work is evidenced by the growing number of publications in which advanced bioreactors are used for in

Tapered bed bioreactor

DOEpatents

Scott, Charles D.; Hancher, Charles W.

1977-01-01

A vertically oriented conically shaped column is used as a fluidized bed bioreactor wherein biologically catalyzed reactions are conducted in a continuous manner. The column utilizes a packing material a support having attached thereto a biologically active catalytic material.

Design concepts for bioreactors in space

NASA Technical Reports Server (NTRS)

Seshan, P. K.; Peterson, G. R.; Beard, B.; Dunlop, E. H.

1986-01-01

Microbial food sources are becoming viable and more efficient alternatives to conventional food sources especially in the context of Closed Ecological Life Support Systems (CELSS) in space habitats. Since bioreactor designs for terrestrial operation will not readily apply to conditions of microgravity, there is an urgent need to learn about the differences. These differences cannot be easily estimated due to the complex nature of the mass transport and mixing mechanisms in fermenters. Therefore, a systematic and expeditious experimental program must be undertaken to obtain the engineering data necessary to lay down the foundations of designing bioreactors for microgravity. Two bioreactor design concepts presented represent two dissimilar approaches to grappling with the absence of gravity in space habitats and deserve to be tested for adoption as important components of the life support function aboard spacecrafts, space stations and other extra-terrestrial habitats.

Fluid mechanics of spinner-flask bioreactors

NASA Astrophysics Data System (ADS)

Sucosky, Philippe; Neitzel, G. Paul

2000-11-01

The dynamic environment within bioreactors used for in vitro tissue growth has been observed to affect the development of mammalian cells. Many studies have shown that moderate mechanical stress enhances growth of some tissues whereas high shear levels and turbulence seem to damage cells. In order to optimize the design and the operating conditions of bioreactors, it is important to understand the fluid-dynamic characteristics and to control the stress levels within these devices. The present research focuses on the characterization of the flow field within a spinner-flask bioreactor. The dynamic properties of the flow are investigated experimentally using particle-image velocimetry with a refractive-index-matched model. Phase-locked ensemble-averaging is employed to provide some information on the turbulence characteristics of the model culture medium in the vicinity of a model tissue construct.

Nitrate Removal Rates in Denitrifying Bioreactors During Storm Flows

NASA Astrophysics Data System (ADS)

Pluer, W.; Walter, T.

2017-12-01

Field denitrifying bioreactors are designed to reduce excess nitrate (NO3-) pollution in runoff from agricultural fields. Field bioreactors saturate organic matter to create conditions that facilitate microbial denitrification. Prior studies using steady flow in lab-scale bioreactors showed that a hydraulic retention time (HRT) between 4 and 10 hours was optimal for reducing NO3- loads. However, during storm-induced events, flow rate and actual HRT fluctuate. These fluctuations have the potential to disrupt the system in significant ways that are not captured by the idealized steady-flow HRT models. The goal of this study was to investigate removal rate during dynamic storm flows of variable rates and durations. Our results indicate that storm peak flow and duration were not significant controlling variables. Instead, we found high correlations (p=0.004) in average removal rates between bioreactors displaying a predominantly uniform flow pattern compared with bioreactors that exhibited preferential flow (24.4 and 21.4 g N m-3 d-1, respectively). This suggests that the internal flow patterns are a more significant driver of removal rate than external factors of the storm hydrograph. Designing for flow patterns in addition to theoretical HRT will facilitate complete mixing within the bioreactors. This will help maximize excess NO3- removal during large storm-induced runoff events.

An Optical Oxygen Sensor for Long-Term Continuous Monitoring of Dissolved Oxygen in Perfused Bioreactors

NASA Technical Reports Server (NTRS)

Gao, F. G.; Jeevarajan, A. S.; Anderson, M. M.

2002-01-01

For long-term growth of man1ITlalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to quantitate and control level of DO. Continuous measurement of the amount of DO in the cell culture medium in-line under sterile conditions in NASA's perfused bioreactor requires that the oxygen sensor provide increased sensitivity and be sterilizable and nontoxic. Additionally, long-term cell culture experiments require that the calibration be maintained several weeks or months. Although there are a number of sensors for dissolved oxygen on the market and under development elsewhere, very few meet these stringent conditions. An optical oxygen sensor (BOXY) based on dynamic fluorescent quenching and a pulsed blue LED light source was developed in our laboratory to address these requirements. Tris( 4,7 -diphenyl-l, 1 O-phenanthroline )ruthenium(II) chloride is employed as the fluorescent dye indicator. The sensing element consists of a glass capillary (OD 4.0 mm; ID 2.0 mm) coated internally with a thin layer of the fluorescent dye in silicone matrix and overlayed with a black shielding layer. Irradiation of the sensing element with blue light (blue LED with emission maximum at 475 nm) generates a red fluorescence centered at 626 nm. The fluorescence intensity is correlated to the concentration of DO present in the culture medium, following the modified non-linear Stern-Volmer equation. By using a pulsed irradiating light source, the problem of dye-bleaching, which is often encountered in long-term continuous measurements of tIns type, 'is minimized. To date we achieved sensor resolution of 0.3 mmHg at 50 mmHg p02, and 0.6 mmHg at 100 mmHg p02, with a response time of about one minute. Calibration was accomplished in sterile phosphate-buffered saline with a blood-gas analyzer (BGA) measurement as reference. Stand-alone software was also developed to control the sensor and bioreactor as well as to

High-density mammalian cell cultures in stirred-tank bioreactor without external pH control.

PubMed

Xu, Sen; Chen, Hao

2016-08-10

Maintaining desired pH is a necessity for optimal cell growth and protein production. It is typically achieved through a two-sided pH control loop on the bioreactor controller. Here we investigated cell culture processes with minimum or no pH control and demonstrated that high-density mammalian cell cultures could be maintained for long-term protein production without pH control. The intrinsic interactions between pCO2, lactate, and pH were leveraged to maintain culture pH. Fed-batch cultures at the same lower pH limit of 6.75 but different upper pH limits (7.05, 7.30, 7.45, 7.65) were evaluated in the 3L bioreactors and comparable results were obtained. Neither CO2 sparging nor base addition was required to control pH in the pH range of 6.75-7.65. The impact of sparger configurations (drilled hole sparger vs. frit sparger) and scales (3L vs. 200L) on CO2 accumulation and culture pH was also demonstrated. The same principle was applied in two perfusion cultures with steady state cell densities at 42.5±3.3 or 68.3±6.0×10(6)cells/mL with low cell specific perfusion rates (15±2 to 23±3pL/cell/day), achieving up to 1.9±0.1g/L/day bioreactor productivity. Culture pH level in the 3L perfusion bioreactors was steadily maintained by controlling the residual lactate and pCO2 levels without the requirement of external pH control for up to 40days with consistent productivity and product quality. Furthermore, culture pH could be potentially modulated via adjusting residual glucose levels and CO2 stripping capability in perfusion cultures. To the best of our knowledge, this is the first time a systematic study was performed to evaluate the long-term cell cultivation and protein production in stirred-tank bioreactors without external pH control. Copyright © 2016 Elsevier B.V. All rights reserved.

Two Devices for Removing Sludge From Bioreactor Wastewater

NASA Technical Reports Server (NTRS)

Archer, Shivaun; Hitchens, G. DUncan; Jabs, Harry; Cross, Jennifer; Pilkinton, Michelle; Taylor, Michael

2007-01-01

Two devices a magnetic separator and a special filter denoted a self-regenerating separator (SRS) have been developed for separating sludge from the stream of wastewater from a bioreactor. These devices were originally intended for use in microgravity, but have also been demonstrated to function in normal Earth gravity. The magnetic separator (see Figure 1) includes a thin-walled nonmagnetic, stainless-steel cylindrical drum that rotates within a cylindrical housing. The wastewater enters the separator through a recirculation inlet, and about 80 percent of the wastewater flow leaves through a recirculation outlet. Inside the drum, a magnet holder positions strong permanent magnets stationary and, except near a recirculation outlet, close to the inner drum surface. To enable magnetic separation, magnetite (a ferromagnetic and magnetically soft iron oxide) powder is mixed into the bioreactor wastewater. The magnetite becomes incorporated into the sludge by condensation, onto the powder particles, of microbe flocks that constitute the sludge. As a result, the magnets inside the drum magnetically attract the sludge onto the outer surface of the drum.

Bacterial Community Dynamics during Start-Up of a Trickle-Bed Bioreactor Degrading Aromatic Compounds

PubMed Central

Stoffels, Marion; Amann, Rudolf; Ludwig, Wolfgang; Hekmat, Dariusch; Schleifer, Karl-Heinz

1998-01-01

This study was performed with a laboratory-scale fixed-bed bioreactor degrading a mixture of aromatic compounds (Solvesso100). The starter culture for the bioreactor was prepared in a fermentor with a wastewater sample of a car painting facility as the inoculum and Solvesso100 as the sole carbon source. The bacterial community dynamics in the fermentor and the bioreactor were examined by a conventional isolation procedure and in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotides. Two significant shifts in the bacterial community structure could be demonstrated. The original inoculum from the wastewater of the car factory was rich in proteobacteria of the alpha and beta subclasses, while the final fermentor enrichment was dominated by bacteria closely related to Pseudomonas putida or Pseudomonas mendocina, which both belong to the gamma subclass of the class Proteobacteria. A second significant shift was observed when the fermentor culture was transferred as inoculum to the trickle-bed bioreactor. The community structure in the bioreactor gradually returned to a higher complexity, with the dominance of beta and alpha subclass proteobacteria, whereas the gamma subclass proteobacteria sharply declined. Obviously, the preceded pollutant adaptant did not lead to a significant enrichment of bacteria that finally dominated in the trickle-bed bioreactor. In the course of experiments, three new 16S as well as 23S rRNA-targeted probes for beta subclass proteobacteria were designed, probe SUBU1237 for the genera Burkholderia and Sutterella, probe ALBO34a for the genera Alcaligenes and Bordetella, and probe Bcv13b for Burkholderia cepacia and Burkholderia vietnamiensis. Bacteria hybridizing with the probe Bcv13b represented the main Solvesso100-degrading population in the reactor. PMID:9501433

Evaluation of woodchip bioreactors for improved water quality

USDA-ARS?s Scientific Manuscript database

Woodchip bioreactors are gaining popularity with farmers because of their edge-of-field nitrate removal capabilities, which do not require changes in land management practices. However, limited research has been conducted to study the potential of these bioreactors to also reduce downstream transpor...

Containerized Wetland Bioreactor Evaluated for Perchlorate and Nitrate Degradation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dibley, V R; Krauter, P W

2004-12-02

The U.S. Department of Energy (DOE) and Lawrence Livermore Laboratory (LLNL) designed and constructed an innovative containerized wetlands (bioreactor) system that began operation in November 2000 to biologically degrade perchlorate and nitrate under relatively low-flow conditions at a remote location at Site 300 known as Building 854. Since initial start-up, the system has processed over 3,463,000 liters of ground water and treated over 38 grams of perchlorate and 148 kilograms of nitrate. Site 300 is operated by the University of California as a high-explosives and materials testing facility supporting nuclear weapons research. The 11-square mile site located in northern Californiamore » was added to the NPL in 1990 primarily due to the presence of elevated concentrations of volatile organic compounds (VOCs) in ground water. At the urging of the regulatory agencies, perchlorate was looked for and detected in the ground water in 1999. VOCs, nitrate and perchlorate were released into the soil and ground water in the Building 854 area as the result of accidental leaks during stability testing of weapons or from waste discharge practices that are no longer permitted at Site 300. Design of the wetland bioreactors was based on earlier studies showing that indigenous chlorate-respiring bacteria could effectively degrade perchlorate into nontoxic concentrations of chlorate, chlorite, oxygen, and chloride. Studies also showed that the addition of organic carbon would enhance microbial denitrification. Early onsite testing showed acetic acid to be a more effective carbon source than dried leaf matter, dried algae, or milk replacement starter; a nutrient and carbon source used in a Department of Defense phytoremediation demonstration. No inocula were added to the system. Groundwater was allowed to circulate through the bioreactor for three weeks to acclimate the wetland plants and to build a biofilm from indigenous flora. Using solar energy, ground water is pumped into

NASA Applied Sciences Program. Overview Presentation; Discovering and Demonstrating Innovative and Practical Applications of Earth Science

NASA Technical Reports Server (NTRS)

Irwin, Daniel

2010-01-01

Goal 1: Enhance Applications Research Advance the use of NASA Earth science in policy making, resource management and planning, and disaster response. Key Actions: Identify priority needs, conduct applied research to generate innovative applications, and support projects that demonstrate uses of NASA Earth science. Goal 2: Increase Collaboration Establish a flexible program structure to meet diverse partner needs and applications objectives. Key Actions: Pursue partnerships to leverage resources and risks and extend the program s reach and impact. Goal 3:Accelerate Applications Ensure that NASA s flight missions plan for and support applications goals in conjunction with their science goals, starting with mission planning and extending through the mission life cycle. Key Actions: Enable identification of applications early in satellite mission lifecycle and facilitate effective ways to integrate end-user needs into satellite mission planning

Characterization of a novel bioreactor system for 3D cellular mechanobiology studies.

PubMed

Cook, Colin A; Huri, Pinar Y; Ginn, Brian P; Gilbert-Honick, Jordana; Somers, Sarah M; Temple, Joshua P; Mao, Hai-Quan; Grayson, Warren L

2016-08-01

In vitro engineering systems can be powerful tools for studying tissue development in response to biophysical stimuli as well as for evaluating the functionality of engineered tissue grafts. It has been challenging, however, to develop systems that adequately integrate the application of biomimetic mechanical strain to engineered tissue with the ability to assess functional outcomes in real time. The aim of this study was to design a bioreactor system capable of real-time conditioning (dynamic, uniaxial strain, and electrical stimulation) of centimeter-long 3D tissue engineered constructs simultaneously with the capacity to monitor local strains. The system addresses key limitations of uniform sample loading and real-time imaging capabilities. Our system features an electrospun fibrin scaffold, which exhibits physiologically relevant stiffness and uniaxial alignment that facilitates cell adhesion, alignment, and proliferation. We have demonstrated the capacity for directly incorporating human adipose-derived stromal/stem cells into the fibers during the electrospinning process and subsequent culture of the cell-seeded constructs in the bioreactor. The bioreactor facilitates accurate pre-straining of the 3D constructs as well as the application of dynamic and static uniaxial strains while monitoring bulk construct tensions. The incorporation of fluorescent nanoparticles throughout the scaffolds enables in situ monitoring of local strain fields using fluorescent digital image correlation techniques, since the bioreactor is imaging compatible, and allows the assessment of local sample stiffness and stresses when coupled with force sensor measurements. In addition, the system is capable of measuring the electromechanical coupling of skeletal muscle explants by applying an electrical stimulus and simultaneously measuring the force of contraction. The packaging of these technologies, biomaterials, and analytical methods into a single bioreactor system has produced a

Open source software to control Bioflo bioreactors.

PubMed

Burdge, David A; Libourel, Igor G L

2014-01-01

Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

Open Source Software to Control Bioflo Bioreactors

PubMed Central

Burdge, David A.; Libourel, Igor G. L.

2014-01-01

Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW. PMID:24667828

Microgravity

NASA Image and Video Library

1998-01-01

Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Fecal Bacteria, Bacteriophage, and Nutrient Reductions in a Full-Scale Denitrifying Woodchip Bioreactor.

PubMed

Rambags, Femke; Tanner, Chris C; Stott, Rebecca; Schipper, Louis A

2016-05-01

Denitrifying bioreactors using woodchips or other slow-release carbon sources can be an effective method for removing nitrate (NO) from wastewater and tile drainage. However, the ability of these systems to remove fecal microbes from wastewater has been largely uninvestigated. In this study, reductions in fecal indicator bacteria () and viruses (F-specific RNA bacteriophage [FRNA phage]) were analyzed by monthly sampling along a longitudinal transect within a full-scale denitrifying woodchip bioreactor receiving secondary-treated septic tank effluent. Nitrogen, phosphorus, 5-d carbonaceous biochemical oxygen demand (CBOD), and total suspended solids (TSS) reduction were also assessed. The bioreactor demonstrated consistent and substantial reduction of (2.9 log reduction) and FRNA phage (3.9 log reduction) despite receiving highly fluctuating inflow concentrations [up to 3.5 × 10 MPN (100 mL) and 1.1 × 10 plaque-forming units (100 mL) , respectively]. Most of the removal of fecal microbial contaminants occurred within the first meter of the system (1.4 log reduction for ; 1.8 log reduction for FRNA phage). The system was also efficient at removing NO (>99.9% reduction) and TSS (89% reduction). There was no evidence of consistent removal of ammonium, organic nitrogen, or phosphorus. Leaching of CBOD occurred during initial operation but decreased and stabilized at lower values (14 g O m) after 9 mo. We present strong evidence for reliable microbial contaminant removal in denitrifying bioreactors, demonstrating their broader versatility for wastewater treatment. Research on the removal mechanisms of microbial contaminants in these systems, together with the assessment of longevity of removal, is warranted. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Dual-Purpose Bioreactors to Monitor Noninvasive Physical and Biochemical Markers of Kidney and Liver Scaffold Recellularization

PubMed Central

Uzarski, Joseph S.; Bijonowski, Brent M.; Wang, Bo; Ward, Heather H.; Wandinger-Ness, Angela

2015-01-01

Analysis of perfusion-based bioreactors for organ engineering and a detailed evaluation of physical and biochemical parameters that measure dynamic changes within maturing cell-laden scaffolds are critical components of ex vivo tissue development that remain understudied topics in the tissue and organ engineering literature. Intricately designed bioreactors that house developing tissue are critical to properly recapitulate the in vivo environment, deliver nutrients within perfused media, and monitor physiological parameters of tissue development. Herein, we provide an in-depth description and analysis of two dual-purpose perfusion bioreactors that improve upon current bioreactor designs and enable comparative analyses of ex vivo scaffold recellularization strategies and cell growth performance during long-term maintenance culture of engineered kidney or liver tissues. Both bioreactors are effective at maximizing cell seeding of small-animal organ scaffolds and maintaining cell survival in extended culture. We further demonstrate noninvasive monitoring capabilities for tracking dynamic changes within scaffolds as the native cellular component is removed during decellularization and model human cells are introduced into the scaffold during recellularization and proliferate in maintenance culture. We found that hydrodynamic pressure drop (ΔP) across the retained scaffold vasculature is a noninvasive measurement of scaffold integrity. We further show that ΔP, and thus resistance to fluid flow through the scaffold, decreases with cell loss during decellularization and correspondingly increases to near normal values for whole organs following recellularization of the kidney or liver scaffolds. Perfused media may be further sampled in real time to measure soluble biomarkers (e.g., resazurin, albumin, or kidney injury molecule-1) that indicate degree of cellular metabolic activity, synthetic function, or engraftment into the scaffold. Cell growth within bioreactors is

Dual-Purpose Bioreactors to Monitor Noninvasive Physical and Biochemical Markers of Kidney and Liver Scaffold Recellularization.

PubMed

Uzarski, Joseph S; Bijonowski, Brent M; Wang, Bo; Ward, Heather H; Wandinger-Ness, Angela; Miller, William M; Wertheim, Jason A

2015-10-01

Analysis of perfusion-based bioreactors for organ engineering and a detailed evaluation of physical and biochemical parameters that measure dynamic changes within maturing cell-laden scaffolds are critical components of ex vivo tissue development that remain understudied topics in the tissue and organ engineering literature. Intricately designed bioreactors that house developing tissue are critical to properly recapitulate the in vivo environment, deliver nutrients within perfused media, and monitor physiological parameters of tissue development. Herein, we provide an in-depth description and analysis of two dual-purpose perfusion bioreactors that improve upon current bioreactor designs and enable comparative analyses of ex vivo scaffold recellularization strategies and cell growth performance during long-term maintenance culture of engineered kidney or liver tissues. Both bioreactors are effective at maximizing cell seeding of small-animal organ scaffolds and maintaining cell survival in extended culture. We further demonstrate noninvasive monitoring capabilities for tracking dynamic changes within scaffolds as the native cellular component is removed during decellularization and model human cells are introduced into the scaffold during recellularization and proliferate in maintenance culture. We found that hydrodynamic pressure drop (ΔP) across the retained scaffold vasculature is a noninvasive measurement of scaffold integrity. We further show that ΔP, and thus resistance to fluid flow through the scaffold, decreases with cell loss during decellularization and correspondingly increases to near normal values for whole organs following recellularization of the kidney or liver scaffolds. Perfused media may be further sampled in real time to measure soluble biomarkers (e.g., resazurin, albumin, or kidney injury molecule-1) that indicate degree of cellular metabolic activity, synthetic function, or engraftment into the scaffold. Cell growth within bioreactors is

Disposable Bioreactors for Plant Micropropagation and Mass Plant Cell Culture

NASA Astrophysics Data System (ADS)

Ducos, Jean-Paul; Terrier, Bénédicte; Courtois, Didier

Different types of bioreactors are used at Nestlé R&D Centre - Tours for mass propagation of selected plant varieties by somatic embryogenesis and for large scale culture of plants cells to produce metabolites or recombinant proteins. Recent studies have been directed to cut down the production costs of these two processes by developing disposable cell culture systems. Vegetative propagation of elite plant varieties is achieved through somatic embryogenesis in liquid medium. A pilot scale process has recently been set up for the industrial propagation of Coffea canephora (Robusta coffee). The current production capacity is 3.0 million embryos per year. The pre-germination of the embryos was previously conducted by temporary immersion in liquid medium in 10-L glass bioreactors. An improved process has been developed using a 10-L disposable bioreactor consisting of a bag containing a rigid plastic box ('Box-in-Bag' bioreactor), insuring, amongst other advantages, a higher light transmittance to the biomass due to its horizontal design. For large scale cell culture, two novel flexible plastic-based disposable bioreactors have been developed from 10 to 100 L working volumes, validated with several plant species ('Wave and Undertow' and 'Slug Bubble' bioreactors). The advantages and the limits of these new types of bioreactor are discussed, based mainly on our own experience on coffee somatic embryogenesis and mass cell culture of soya and tobacco.

Nitrate and phosphate removal from agricultural subsurface drainage using laboratory woodchip bioreactors and recycled steel byproduct filters.

PubMed

Hua, Guanghui; Salo, Morgan W; Schmit, Christopher G; Hay, Christopher H

2016-10-01

Woodchip bioreactors have been increasingly used as an edge-of-field treatment technology to reduce the nitrate loadings to surface waters from agricultural subsurface drainage. Recent studies have shown that subsurface drainage can also contribute substantially to the loss of phosphate from agricultural soils. The objective of this study was to investigate nitrate and phosphate removal in subsurface drainage using laboratory woodchip bioreactors and recycled steel byproduct filters. The woodchip bioreactor demonstrated average nitrate removal efficiencies of 53.5-100% and removal rates of 10.1-21.6 g N/m(3)/d for an influent concentration of 20 mg N/L and hydraulic retention times (HRTs) of 6-24 h. When the influent nitrate concentration increased to 50 mg N/L, the bioreactor nitrate removal efficiency and rate averaged 75% and 18.9 g N/m(3)/d at an HRT of 24 h. Nitrate removal by the woodchips followed zero-order kinetics with rate constants of 1.42-1.80 mg N/L/h when nitrate was non-limiting. The steel byproduct filter effectively removed phosphate in the bioreactor effluent and the total phosphate adsorption capacity was 3.70 mg P/g under continuous flow conditions. Nitrite accumulation occurred in the woodchip bioreactor and the effluent nitrite concentrations increased with decreasing HRTs and increasing influent nitrate concentrations. The steel byproduct filter efficiently reduced the level of nitrite in the bioreactor effluent. Overall, the results of this study suggest that woodchip denitrification followed by steel byproduct filtration is an effective treatment technology for nitrate and phosphate removal in subsurface drainage. Published by Elsevier Ltd.

Hollow microcarriers for large-scale expansion of anchorage-dependent cells in a stirred bioreactor.

PubMed

YekrangSafakar, Ashkan; Acun, Aylin; Choi, Jin-Woo; Song, Edward; Zorlutuna, Pinar; Park, Kidong

2018-03-26

With recent advances in biotechnology, mammalian cells are used in biopharmaceutical industries to produce valuable protein therapeutics and investigated as effective therapeutic agents to permanently degenerative diseases in cell based therapy. In these exciting and actively expanding fields, a reliable, efficient, and affordable platform to culture mammalian cells on a large scale is one of the most vital necessities. To produce and maintain a very large population of anchorage-dependent cells, a microcarrier-based stirred tank bioreactor is commonly used. In this approach, the cells are exposed to harmful hydrodynamic shear stress in the bioreactor and the mass transfer rates of nutrients and gases in the bioreactor are often kept below an optimal level to prevent cellular damages from the shear stress. In this paper, a hollow microcarrier (HMC) is presented as a novel solution to protect cells from shear stress in stirred bioreactors, while ensuring sufficient and uniform mass transfer rate of gases and nutrients. HMC is a hollow microsphere and cells are cultured on its inner surface to be protected, while openings on the HMC provide sufficient exchange of media inside the HMC. As a proof of concept, we demonstrated the expansion of fibroblasts, NIH/3T3 and the expansion and cardiac differentiation of human induced pluripotent stem cells, along with detailed numerical analysis. We believe that the developed HMC can be a practical solution to enable large-scale expansion of shear-sensitive anchorage-dependent cells in an industrial scale with stirred bioreactors. © 2018 Wiley Periodicals, Inc.

Upflow bioreactor with septum and pressure release mechanism

DOEpatents

Hansen, Conly L.; Hansen, Carl S.; Pack, Kevin; Milligan, John; Benefiel, Bradley C.; Tolman, C. Wayne; Tolman, Kenneth W.

2010-04-20

An upflow bioreactor includes a vessel having an inlet and an outlet configured for upflow operation. A septum is positioned within the vessel and defines a lower chamber and an upper chamber. The septum includes an aperture that provides fluid communication between the upper chamber and lower chamber. The bioreactor also includes means for releasing pressure buildup in the lower chamber. In one configuration, the septum includes a releasable portion having an open position and a closed position. The releasable portion is configured to move to the open position in response to pressure buildup in the lower chamber. In the open position fluid communication between the lower chamber and the upper chamber is increased. Alternatively the lower chamber can include a pressure release line that is selectively actuated by pressure buildup. The pressure release mechanism can prevent the bioreactor from plugging and/or prevent catastrophic damage to the bioreactor caused by high pressures.

Fluidized-bed bioreactor process for the microbial solubiliztion of coal

DOEpatents

Scott, Charles D.; Strandberg, Gerald W.

1989-01-01

A fluidized-bed bioreactor system for the conversion of coal into microbially solubilized coal products. The fluidized-bed bioreactor continuously or periodically receives coal and bio-reactants and provides for the production of microbially solubilized coal products in an economical and efficient manner. An oxidation pretreatment process for rendering coal uniformly and more readily susceptible to microbial solubilization may be employed with the fluidized-bed bioreactor.

Simplified Bioreactor For Growing Mammalian Cells

NASA Technical Reports Server (NTRS)

Spaulding, Glenn F.

1995-01-01

Improved bioreactor for growing mammalian cell cultures developed. Designed to support growth of dense volumes of mammalian cells by providing ample, well-distributed flows of nutrient solution with minimal turbulence. Cells relatively delicate and, unlike bacteria, cannot withstand shear forces present in turbulent flows. Bioreactor vessel readily made in larger sizes to accommodate greater cell production quantities. Molding equipment presently used makes cylinders up to 30 centimeters long. Alternative sintered plastic techniques used to vary pore size and quantity, as necessary.

Three-dimensional neural differentiation of embryonic stem cells with ACM induction in microfibrous matrices in bioreactors.

PubMed

Liu, Ning; Ouyang, Anli; Li, Yan; Yang, Shang-Tian

2013-01-01

The clinical use of pluripotent stem cell (PSC)-derived neural cells requires an efficient differentiation process for mass production in a bioreactor. Toward this goal, neural differentiation of murine embryonic stem cells (ESCs) in three-dimensional (3D) polyethylene terephthalate microfibrous matrices was investigated in this study. To streamline the process and provide a platform for process integration, the neural differentiation of ESCs was induced with astrocyte-conditioned medium without the formation of embryoid bodies, starting from undifferentiated ESC aggregates expanded in a suspension bioreactor. The 3D neural differentiation was able to generate a complex neural network in the matrices. When compared to 2D differentiation, 3D differentiation in microfibrous matrices resulted in a higher percentage of nestin-positive cells (68% vs. 54%) and upregulated gene expressions of nestin, Nurr1, and tyrosine hydroxylase. High purity of neural differentiation in 3D microfibrous matrix was also demonstrated in a spinner bioreactor with 74% nestin + cells. This study demonstrated the feasibility of a scalable process based on 3D differentiation in microfibrous matrices for the production of ESC-derived neural cells. © 2013 American Institute of Chemical Engineers.

Multimembrane Bioreactor

NASA Technical Reports Server (NTRS)

Cho, Toohyon; Shuler, Michael L.

1989-01-01

Set of hydrophilic and hydrophobic membranes in bioreactor allows product of reaction to be separated, while nutrients fed to reacting cells and byproducts removed from them. Separation process requires no externally supplied energy; free energy of reaction sufficient. Membranes greatly increase productivity of metabolizing cells by continuously removing product and byproducts, which might otherwise inhibit reaction, and by continuously adding oxygen and organic nutrients.

Scale up of diesel oil biodegradation in a baffled roller bioreactor.

PubMed

Nikakhtari, Hossein; Song, Wanning; Kumar, Pardeep; Nemati, Mehdi; Hill, Gordon A

2010-05-01

Diesel oil is a suitable substance to represent petroleum contamination from accidental spills in operating and transportation facilities. Using a microbial culture enriched from a petroleum contaminated soil, biodegradation of diesel oil was carried out in 2.2, 55, and 220 L roller baffled bioreactors. The effects of bioreactor rotation speed (from 5 to 45 rpm) and liquid loading (from 18% to 73% of total volume) on the biodegradation of diesel oil were studied. In the small scale bioreactor (2.2L), the maximum rotation speed of 45 rpm resulted in the highest biodegradation rate with a first order biodegradation kinetic constant of 0.095 d(-1). In the larger scale bioreactors, rotation speed did not affect the biodegradation rate. Liquid loadings higher than 64% resulted in reduced biodegradation rates in the small scale bioreactor; however, in the larger roller bioreactors liquid loading did not affect the biodegradation rate. Biodegradation of diesel oil at 5 rpm and 73% loading is recommended for operating large scale roller baffled bioreactors. Under these conditions, high diesel oil concentrations up to 50 gL(-1) can be bioremediated at a rate of 1.61 gL(-1)d(-1). Copyright 2010 Elsevier Ltd. All rights reserved.

Fluidized-bed bioreactor system for the microbial solubilization of coal

DOEpatents

Scott, C.D.; Strandberg, G.W.

1987-09-14

A fluidized-bed bioreactor system for the conversion of coal into microbially solubilized coal products. The fluidized-bed bioreactor continuously or periodically receives coal and bio-reactants and provides for the production of microbially solubilized coal products in an economical and efficient manner. An oxidation pretreatment process for rendering coal uniformly and more readily susceptible to microbial solubilization may be employed with the fluidized-bed bioreactor. 2 figs.

[Study of shear rate in modified airlift nitrifying bioreactor].

PubMed

Jin, Ren-cun; Zheng, Ping

2006-06-01

The characteristics of shear rate in an airlift nitrifying bioreactor and its influencing factors were studied. The results showed that the shear rate was different in different sections of the bioreactor. With inlet gas flowrate at 430 approximately 2700 L x h(-1), the overall shear rate was (0.702 approximately 3.13) x 10(5) s(-1), shear rate in riser was (1.07 approximately 31.3) x 10(5) s(-1) and in gas-liquid separator was (1.12 approximately 25.0) x 10(5) s(-1), respectively. It indicates that the highest shear rates prevailed in the riser part of bioreactor. The operational variables and the bioreactor configurations exerted a significant influence on the shear level of the bioreactor. When inlet gas flowrate was raised from 1300 to 2700 L x h(-1), shear rate in riser and separator ascended first and then descended subsequently. The diameter of draft tube (d) was negatively correlated with shear rate. When the draft tube with diameter of 5.5 cm was installed, the shear rates in riser, separator and overall shear rate were 85.5%, 82.3% and 80.6%, respectively less as compared with that with diameter of 4.0 cm. The number of static mixers (N) was positively correlated with the shear rate. When d was set at 4.0 cm, with N of 10 and 39, the shear rates in riser were 6.14 and 7.97 times higher respectively, than that of conventional bioreactor. The ratio of maximum local shear rate to overall shear rate was 3.68 approximately 7.66, and the homogeneity of the shear field in airlift bioreactors could be improved if d and N were set at 5.5 cm and 10 approximately 13, respectively.

Oscillating Cell Culture Bioreactor

NASA Technical Reports Server (NTRS)

Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

2010-01-01

To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

Simulation of municipal solid waste degradation in aerobic and anaerobic bioreactor landfills.

PubMed

Patil, Bhagwan Shamrao; C, Agnes Anto; Singh, Devendra Narain

2017-03-01

Municipal solid waste generation is huge in growing cities of developing nations such as India, owing to the rapid industrial and population growth. In addition to various methods for treatment and disposal of municipal solid waste (landfills, composting, bio-methanation, incineration and pyrolysis), aerobic/anaerobic bioreactor landfills are gaining popularity for economical and effective disposal of municipal solid waste. However, efficiency of municipal solid waste bioreactor landfills primarily depends on the municipal solid waste decomposition rate, which can be accelerated through monitoring moisture content and temperature by using the frequency domain reflectometry probe and thermocouples, respectively. The present study demonstrates that these landfill physical properties of the heterogeneous municipal solid waste mass can be monitored using these instruments, which facilitates proper scheduling of the leachate recirculation for accelerating the decomposition rate of municipal solid waste.

Large Scale Expansion of Human Umbilical Cord Cells in a Rotating Bed System Bioreactor for Cardiovascular Tissue Engineering Applications

PubMed Central

Reichardt, Anne; Polchow, Bianca; Shakibaei, Mehdi; Henrich, Wolfgang; Hetzer, Roland; Lueders, Cora

2013-01-01

Widespread use of human umbilical cord cells for cardiovascular tissue engineering requires production of large numbers of well-characterized cells under controlled conditions. In current research projects, the expansion of cells to be used to create a tissue construct is usually performed in static cell culture systems which are, however, often not satisfactory due to limitations in nutrient and oxygen supply. To overcome these limitations dynamic cell expansion in bioreactor systems under controllable conditions could be an important tool providing continuous perfusion for the generation of large numbers of viable pre-conditioned cells in a short time period. For this purpose cells derived from human umbilical cord arteries were expanded in a rotating bed system bioreactor for up to 9 days. For a comparative study, cells were cultivated under static conditions in standard culture devices. Our results demonstrated that the microenvironment in the perfusion bioreactor was more favorable than that of the standard cell culture flasks. Data suggested that cells in the bioreactor expanded 39 fold (38.7 ± 6.1 fold) in comparison to statically cultured cells (31.8 ± 3.0 fold). Large-scale production of cells in the bioreactor resulted in more than 3 x 108 cells from a single umbilical cord fragment within 9 days. Furthermore cell doubling time was lower in the bioreactor system and production of extracellular matrix components was higher. With this study, we present an appropriate method to expand human umbilical cord artery derived cells with high cellular proliferation rates in a well-defined bioreactor system under GMP conditions. PMID:23847691

Biomimetic fetal rotation bioreactor for engineering bone tissues-Effect of cyclic strains on upregulation of osteogenic gene expression.

PubMed

Ravichandran, Akhilandeshwari; Wen, Feng; Lim, Jing; Chong, Mark Seow Khoon; Chan, Jerry K Y; Teoh, Swee-Hin

2018-04-01

Cells respond to physiological mechanical stresses especially during early fetal development. Adopting a biomimetic approach, it is necessary to develop bioreactor systems to explore the effects of physiologically relevant mechanical strains and shear stresses for functional tissue growth and development. This study introduces a multimodal bioreactor system that allows application of cyclic compressive strains on premature bone grafts that are cultured under biaxial rotation (chamber rotation about 2 axes) conditions for bone tissue engineering. The bioreactor is integrated with sensors for dissolved oxygen levels and pH that allow real-time, non-invasive monitoring of the culture parameters. Mesenchymal stem cells-seeded polycaprolactone-β-tricalcium phosphate scaffolds were cultured in this bioreactor over 2 weeks in 4 different modes-static, cyclic compression, biaxial rotation, and multimodal (combination of cyclic compression and biaxial rotation). The multimodal culture resulted in 1.8-fold higher cellular proliferation in comparison with the static controls within the first week. Two weeks of culture in the multimodal bioreactor utilizing the combined effects of optimal fluid flow conditions and cyclic compression led to the upregulation of osteogenic genes alkaline phosphatase (3.2-fold), osteonectin (2.4-fold), osteocalcin (10-fold), and collagen type 1 α1 (2-fold) in comparison with static cultures. We report for the first time, the independent and combined effects of mechanical stimulation and biaxial rotation for bone tissue engineering using a bioreactor platform with non-invasive sensing modalities. The demonstrated results show leaning towards the futuristic vision of using a physiologically relevant bioreactor system for generation of autologous bone grafts for clinical implantation. Copyright © 2018 John Wiley & Sons, Ltd.

NASA/ASEE Summer Faculty Fellowship Program, 1990, Volume 1

NASA Technical Reports Server (NTRS)

Bannerot, Richard B. (Editor); Goldstein, Stanley H. (Editor)

1990-01-01

The 1990 Johnson Space Center (JSC) NASA/American Society for Engineering Education (ASEE) Summer Faculty Fellowship Program was conducted by the University of Houston-University Park and JSC. A compilation of the final reports on the research projects are presented. The topics covered include: the Space Station; the Space Shuttle; exobiology; cell biology; culture techniques; control systems design; laser induced fluorescence; spacecraft reliability analysis; reduced gravity; biotechnology; microgravity applications; regenerative life support systems; imaging techniques; cardiovascular system; physiological effects; extravehicular mobility units; mathematical models; bioreactors; computerized simulation; microgravity simulation; and dynamic structural analysis.

Overview of the Solar Dynamic Ground Test Demonstration Program at the NASA Lewis Research Center

NASA Technical Reports Server (NTRS)

Shaltens, Richard K.

1995-01-01

The Solar Dynamic (SD) Ground Test Demonstration (GTD) program demonstrates the availability of SD technologies in a simulated space environment at the NASA Lewis Research Center (LERC) vacuum facility. Data from the SD GTD program will be provided to the joint U.S. and Russian team which is currently designing a 2 kW SD flight demonstration power system. This SD technology has the potential as a future power source for the International Space Station. This paper reviews the goals and status of the SD GTD program. A description of the SD GTD system includes key design features of the system, subsystems and components.

Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells.

PubMed

Mitra, Debika; Whitehead, Jacklyn; Yasui, Osamu W; Leach, J Kent

2017-11-01

Perfusion culture of mesenchymal stem cells (MSCs) seeded in biomaterial scaffolds provides nutrients for cell survival, enhances extracellular matrix deposition, and increases osteogenic cell differentiation. However, there is no consensus on the appropriate perfusion duration of cellular constructs in vitro to boost their bone forming capacity in vivo. We investigated this phenomenon by culturing human MSCs in macroporous composite scaffolds in a direct perfusion bioreactor and compared their response to scaffolds in continuous dynamic culture conditions on an XYZ shaker. Cell seeding in continuous perfusion bioreactors resulted in more uniform MSC distribution than static seeding. We observed similar calcium deposition in all composite scaffolds over 21 days of bioreactor culture, regardless of pore size. Compared to scaffolds in dynamic culture, perfused scaffolds exhibited increased DNA content and expression of osteogenic markers up to 14 days in culture that plateaued thereafter. We then evaluated the effect of perfusion culture duration on bone formation when MSC-seeded scaffolds were implanted in a murine ectopic site. Human MSCs persisted in all scaffolds at 2 weeks in vivo, and we observed increased neovascularization in constructs cultured under perfusion for 7 days relative to those cultured for 1 day within each gender. At 8 weeks post-implantation, we observed greater bone volume fraction, bone mineral density, tissue ingrowth, collagen density, and osteoblastic markers in bioreactor constructs cultured for 14 days compared to those cultured for 1 or 7 days, and acellular constructs. Taken together, these data demonstrate that culturing MSCs under perfusion culture for at least 14 days in vitro improves the quantity and quality of bone formation in vivo. This study highlights the need for optimizing in vitro bioreactor culture duration of engineered constructs to achieve the desired level of bone formation. Copyright © 2017 Elsevier Ltd. All

Denitrifying bioreactors for nitrate removal from tile drained cropland

USDA-ARS?s Scientific Manuscript database

Denitrification bioreactors are a promising technology for mitigation of nitrate-nitrogen (NO3-N) losses in subsurface drainage water. Bioreactors are constructed with carbon substrates, typically wood chips, to provide a substrate for denitrifying microorganisms. Researchers in Iowa found that for ...

ADVANCING THE FIELD EVALUATIONS AND APPLICATIONS OF LANDFILL BIOREACTORS

EPA Science Inventory

The US Environmental Protection Agency (EPA) is undertaking a long-term program to conduct field evaluations of landfill bioreactors. The near-term effort is focused on the development of appropriate monitoring strategies to ensure adequate control of the landfill bioreactors an...

Fixed-bed bioreactor system for the microbial solubilization of coal

DOEpatents

Scott, C.D.; Strandberg, G.W.

1987-09-14

A fixed-bed bioreactor system for the conversion of coal into microbially solubilized coal products. The fixed-bed bioreactor continuously or periodically receives coal and bio-reactants and provides for the large scale production of microbially solubilized coal products in an economical and efficient manner. An oxidation pretreatment process for rendering coal uniformly and more readily susceptible to microbial solubilization may be employed with the fixed-bed bioreactor. 1 fig., 1 tab.

Modular bioreactor for the remediation of liquid streams and methods for using the same

DOEpatents

Noah, Karl S.; Sayer, Raymond L.; Thompson, David N.

1998-01-01

The present invention is directed to a bioreactor system for the remediation of contaminated liquid streams. The bioreactor system is composed of at least one and often a series of sub-units referred to as bioreactor modules. The modular nature of the system allows bioreactor systems be subdivided into smaller units and transported to waste sites where they are combined to form bioreactor systems of any size. The bioreactor modules further comprises reactor fill materials in the bioreactor module that remove the contaminants from the contaminated stream. To ensure that the stream thoroughly contacts the reactor fill materials, each bioreactor module comprises means for directing the flow of the stream in a vertical direction and means for directing the flow of the stream in a horizontal direction. In a preferred embodiment, the reactor fill comprises a sulfate reducing bacteria which is particularly useful for precipitating metals from acid mine streams.

Plantform Bioreactor for Mass Micropropagation of Date Palm.

PubMed

Almusawi, Abdulminam H A; Sayegh, Abdullah J; Alshanaw, Ansam M S; Griffis, John L

2017-01-01

A novel protocol for the commercial production of date palm through micropropagation is presented. This protocol includes the use of a semisolid medium alternation or in combination with a temporary immersion system (TIS, Plantform bioreactor) in date palm micropropagation. The use of the Plantform bioreactor for date palm results in an improved multiplication rate, reduced micropropagation time, and improved weaning success. It also reduces the cost of saleable units and thus improves economic return for commercial micropropagation. The use of the Plantform bioreactor successfully addresses other hindrances that can occur during the scale-up of date palm micropropagation, including asynchrony of somatic embryos, limited maturation of somatic embryos, and highly variable germination frequencies of embryos.

Plant cell cultures: bioreactors for industrial production.

PubMed

Ruffoni, Barbara; Pistelli, Laura; Bertoli, Alessandra; Pistelli, Luisa

2010-01-01

The recent biotechnology boom has triggered increased interest in plant cell cultures, since a number of firms and academic institutions investigated intensively to rise the production of very promising bioactive compounds. In alternative to wild collection or plant cultivation, the production of useful and valuable secondary metabolites in large bioreactors is an attractive proposal; it should contribute significantly to future attempts to preserve global biodiversity and alleviate associated ecological problems. The advantages of such processes include the controlled production according to demand and a reduced man work requirement. Plant cells have been grown in different shape bioreactors, however, there are a variety of problems to be solved before this technology can be adopted on a wide scale for the production of useful plant secondary metabolites. There are different factors affecting the culture growth and secondary metabolite production in bioreactors: the gaseous atmosphere, oxygen supply and CO2 exchange, pH, minerals, carbohydrates, growth regulators, the liquid medium rheology and cell density. Moreover agitation systems and sterilization conditions may negatively influence the whole process. Many types ofbioreactors have been successfully used for cultivating transformed root cultures, depending on both different aeration system and nutrient supply. Several examples of medicinal and aromatic plant cultures were here summarized for the scale up cultivation in bioreactors.

Continuous pH monitoring in a perfused bioreactor system using an optical pH sensor

NASA Technical Reports Server (NTRS)

Jeevarajan, Antony S.; Vani, Sundeep; Taylor, Thomas D.; Anderson, Melody M.

2002-01-01

Monitoring and regulating the pH of the solution in a bioprocess is one of the key steps in the success of bioreactor operation. An in-line optical pH sensor, based on the optical absorption properties of phenol red present in the medium, was developed and tested in this work for use in NASA space bioreactors based on a rotating wall-perfused vessel system supporting a baby hamster kidney (BHK-21) cell culture. The sensor was tested over three 30-day and one 124-day cell runs. The pH sensor initially was calibrated and then used during the entire cell culture interval. The pH reported by the sensor was compared to that measured by a fiber optically coupled Shimadzu spectrophotometer and a blood gas analyzer. The maximum standard error of prediction for all the four cell runs for development pH sensor against BGA was +/-0.06 pH unit and for the fiber optically coupled Shimadzu spectrophotometer against the blood gas analyzer was +/-0.05 pH unit. The pH sensor system performed well without need of recalibration for 124 days. Copyright 2002 Wiley Periodicals, Inc.

Continuous D-tagatose production by immobilized thermostable L-arabinose isomerase in a packed-bed bioreactor.

PubMed

Ryu, Se-Ah; Kim, Chang Sup; Kim, Hye-Jung; Baek, Dae Heoun; Oh, Deok-Kun

2003-01-01

D-Tagatose was continuously produced using thermostable L-arabinose isomerase immobilized in alginate with D-galactose solution in a packed-bed bioreactor. Bead size, L/D (length/diameter) of reactor, dilution rate, total loaded enzyme amount, and substrate concentration were found to be optimal at 0.8 mm, 520/7 mm, 0.375 h(-1), 5.65 units, and 300 g/L, respectively. Under these conditions, the bioreactor produced about 145 g/L tagatose with an average productivity of 54 g tagatose/L x h and an average conversion yield of 48% (w/w). Operational stability of the immobilized enzyme was demonstrated, with a tagatose production half-life of 24 days.

Pilot demonstration of energy-efficient membrane bioreactor (MBR) using reciprocating submerged membrane.

PubMed

Ho, Jaeho; Smith, Shaleena; Patamasank, Jaren; Tontcheva, Petia; Kim, Gyu Dong; Roh, Hyung Keun

2015-03-01

Membrane bioreactor (MBR) is becoming popular for advanced wastewater treatment and water reuse. Air scouring to "shake" the membrane fibers is most suitable and applicable to maintain filtration without severe and rapidfouling. However, membrane fouling mitigating technologies are energy intensive. The goal of this research is to develop an alternative energy-saving MBR system to reduce energy consumption; a revolutionary system that will directly compete with air scouring technologies currently in the membrane water reuse market. The innovative MBR system, called reciprocation MBR (rMBR), prevents membrane fouling without the use of air scouring blowers. The mechanism featured is a mechanical reciprocating membrane frame that uses inertia to prevent fouling. Direct strong agitation of the fiber is also beneficial for the constant removal of solids built up on the membrane surface. The rMBR pilot consumes less energy than conventional coarse air scouring MBR systems. Specific energy consumption for membrane reciprocation for the pilot rMBR system was 0.072 kWh/m3 permeate produced at 40 LMH, which is 75% less than the conventional air scouring in an MBR system (0.29 kWh/m3). Reciprocation of the hollow-fiber membrane can overcome the hydrodynamic limitations of air scouring or cross-flow membrane systems with less energy consumption and/or higher energy efficiency.

Modular bioreactor for the remediation of liquid streams and methods for using the same

DOEpatents

Noah, K.S.; Sayer, R.L.; Thompson, D.N.

1998-06-30

The present invention is directed to a bioreactor system for the remediation of contaminated liquid streams. The bioreactor system is composed of at least one and often a series of sub-units referred to as bioreactor modules. The modular nature of the system allows bioreactor systems be subdivided into smaller units and transported to waste sites where they are combined to form bioreactor systems of any size. The bioreactor modules further comprises reactor fill materials in the bioreactor module that remove the contaminants from the contaminated stream. To ensure that the stream thoroughly contacts the reactor fill materials, each bioreactor module comprises means for directing the flow of the stream in a vertical direction and means for directing the flow of the stream in a horizontal direction. In a preferred embodiment, the reactor fill comprises a sulfate reducing bacteria which is particularly useful for precipitating metals from acid mine streams. 6 figs.

Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture.

PubMed

Storm, Michael P; Sorrell, Ian; Shipley, Rebecca; Regan, Sophie; Luetchford, Kim A; Sathish, Jean; Webb, Steven; Ellis, Marianne J

2016-05-26

Tissue culture has been used for over 100 years to study cells and responses ex vivo. The convention of this technique is the growth of anchorage dependent cells on the 2-dimensional surface of tissue culture plastic. More recently, there is a growing body of data demonstrating more in vivo-like behaviors of cells grown in 3-dimensional culture systems. This manuscript describes in detail the set-up and operation of a hollow fiber bioreactor system for the in vivo-like culture of mammalian cells. The hollow fiber bioreactor system delivers media to the cells in a manner akin to the delivery of blood through the capillary networks in vivo. The system is designed to fit onto the shelf of a standard CO2 incubator and is simple enough to be set-up by any competent cell biologist with a good understanding of aseptic technique. The systems utility is demonstrated by culturing the hepatocarcinoma cell line HepG2/C3A for 7 days. Further to this and in line with other published reports on the functionality of cells grown in 3-dimensional culture systems the cells are shown to possess increased albumin production (an important hepatic function) when compared to standard 2-dimensional tissue culture.

Staying alive! Sensors used for monitoring cell health in bioreactors.

PubMed

O'Mara, P; Farrell, A; Bones, J; Twomey, K

2018-01-01

Current and next generation sensors such as pH, dissolved oxygen (dO) and temperature sensors that will help drive the use of single-use bioreactors in industry are reviewed. The current trend in bioreactor use is shifting from the traditional fixed bioreactors to the use of single-use bioreactors (SUBs). However as the shift in paradigm occurs there is now a greater need for sensor technology to play 'catch up' with the innovation of bioreactor technology. Many of the sensors still in use today rely on technology created in the 1960's such as the Clark-type dissolved oxygen sensor or glass pH electrodes. This is due to the strict requirements of sensors to monitor bioprocesses resulting in the use of traditional well understood methods, making it difficult to incorporate new sensor technology into industry. A number of advances in sensor technology have been achieved in recent years, a few of these advances and future research will also be discussed in this review. Copyright © 2017 Elsevier B.V. All rights reserved.

Structures and Design Phase I Summary for the NASA Composite Cryotank Technology Demonstration Project

NASA Technical Reports Server (NTRS)

Johnson, Ted; Sleight, David W.; Martin, Robert A.

2013-01-01

A description of the Phase I structures and design work of the Composite Cryotank Technology Demonstration (CCTD) Project is in this paper. The goal of the CCTD Project in the Game Changing Development (GCD) Program is to design and build a composite liquid-hydrogen cryogenic tank that can save 30% in weight and 25% in cost compared to state-of-the-art aluminum metallic cryogenic tank technology when the wetted composite skin wall is at an allowable strain of 5000 in/in. Three Industry teams developed composite cryogenic tank concepts that are compared for weight to an aluminum-lithium (Al-Li) cryogenic tank designed by NASA in Phase I of the CCTD Project. The requirements used to design all of the cryogenic tanks in Phase I will be discussed and the resulting designs, analyses, and weight of the concepts developed by NASA and Industry will be reviewed and compared.

The Use of Multidimensional Image-Based Analysis to Accurately Monitor Cell Growth in 3D Bioreactor Culture

PubMed Central

Baradez, Marc-Olivier; Marshall, Damian

2011-01-01

The transition from traditional culture methods towards bioreactor based bioprocessing to produce cells in commercially viable quantities for cell therapy applications requires the development of robust methods to ensure the quality of the cells produced. Standard methods for measuring cell quality parameters such as viability provide only limited information making process monitoring and optimisation difficult. Here we describe a 3D image-based approach to develop cell distribution maps which can be used to simultaneously measure the number, confluency and morphology of cells attached to microcarriers in a stirred tank bioreactor. The accuracy of the cell distribution measurements is validated using in silico modelling of synthetic image datasets and is shown to have an accuracy >90%. Using the cell distribution mapping process and principal component analysis we show how cell growth can be quantitatively monitored over a 13 day bioreactor culture period and how changes to manufacture processes such as initial cell seeding density can significantly influence cell morphology and the rate at which cells are produced. Taken together, these results demonstrate how image-based analysis can be incorporated in cell quality control processes facilitating the transition towards bioreactor based manufacture for clinical grade cells. PMID:22028809

The use of multidimensional image-based analysis to accurately monitor cell growth in 3D bioreactor culture.

PubMed

Baradez, Marc-Olivier; Marshall, Damian

2011-01-01

The transition from traditional culture methods towards bioreactor based bioprocessing to produce cells in commercially viable quantities for cell therapy applications requires the development of robust methods to ensure the quality of the cells produced. Standard methods for measuring cell quality parameters such as viability provide only limited information making process monitoring and optimisation difficult. Here we describe a 3D image-based approach to develop cell distribution maps which can be used to simultaneously measure the number, confluency and morphology of cells attached to microcarriers in a stirred tank bioreactor. The accuracy of the cell distribution measurements is validated using in silico modelling of synthetic image datasets and is shown to have an accuracy >90%. Using the cell distribution mapping process and principal component analysis we show how cell growth can be quantitatively monitored over a 13 day bioreactor culture period and how changes to manufacture processes such as initial cell seeding density can significantly influence cell morphology and the rate at which cells are produced. Taken together, these results demonstrate how image-based analysis can be incorporated in cell quality control processes facilitating the transition towards bioreactor based manufacture for clinical grade cells.

The Potential for Microalgae as Bioreactors to Produce Pharmaceuticals

PubMed Central

Yan, Na; Fan, Chengming; Chen, Yuhong; Hu, Zanmin

2016-01-01

As photosynthetic organisms, microalgae can efficiently convert solar energy into biomass. Microalgae are currently used as an important source of valuable natural biologically active molecules, such as carotenoids, chlorophyll, long-chain polyunsaturated fatty acids, phycobiliproteins, carotenoids and enzymes. Significant advances have been achieved in microalgae biotechnology over the last decade, and the use of microalgae as bioreactors for expressing recombinant proteins is receiving increased interest. Compared with the bioreactor systems that are currently in use, microalgae may be an attractive alternative for the production of pharmaceuticals, recombinant proteins and other valuable products. Products synthesized via the genetic engineering of microalgae include vaccines, antibodies, enzymes, blood-clotting factors, immune regulators, growth factors, hormones, and other valuable products, such as the anticancer agent Taxol. In this paper, we briefly compare the currently used bioreactor systems, summarize the progress in genetic engineering of microalgae, and discuss the potential for microalgae as bioreactors to produce pharmaceuticals. PMID:27322258

Start-up of membrane bioreactor and hybrid moving bed biofilm reactor-membrane bioreactor: kinetic study.

PubMed

Leyva-Díaz, J C; Poyatos, J M

2015-01-01

A hybrid moving bed biofilm reactor-membrane bioreactor (hybrid MBBR-MBR) system was studied as an alternative solution to conventional activated sludge processes and membrane bioreactors. This paper shows the results obtained from three laboratory-scale wastewater treatment plants working in parallel in the start-up and steady states. The first wastewater treatment plant was a MBR, the second one was a hybrid MBBR-MBR system containing carriers both in anoxic and aerobic zones of the bioreactor (hybrid MBBR-MBRa), and the last one was a hybrid MBBR-MBR system which contained carriers only in the aerobic zone (hybrid MBBR-MBRb). The reactors operated with a hydraulic retention time of 30.40 h. A kinetic study for characterizing heterotrophic biomass was carried out and organic matter and nutrients removals were evaluated. The heterotrophic biomass of the hybrid MBBR-MBRb showed the best kinetic performance in the steady state, with yield coefficient for heterotrophic biomass=0.30246 mg volatile suspended solids per mg chemical oxygen demand, maximum specific growth rate for heterotrophic biomass=0.00308 h(-1) and half-saturation coefficient for organic matter=3.54908 mg O2 L(-1). The removal of organic matter was supported by the kinetic study of heterotrophic biomass.

Gas hold-up and oxygen mass transfer in three pneumatic bioreactors operating with sugarcane bagasse suspensions.

PubMed

Esperança, M N; Cunha, F M; Cerri, M O; Zangirolami, T C; Farinas, C S; Badino, A C

2014-05-01

Sugarcane bagasse is a low-cost and abundant by-product generated by the bioethanol industry, and is a potential substrate for cellulolytic enzyme production. The aim of this work was to evaluate the effects of air flow rate (QAIR), solids loading (%S), sugarcane bagasse type, and particle size on the gas hold-up (εG) and volumetric oxygen transfer coefficient (kLa) in three different pneumatic bioreactors, using response surface methodology. Concentric tube airlift (CTA), split-cylinder airlift (SCA), and bubble column (BC) bioreactor types were tested. QAIR and %S affected oxygen mass transfer positively and negatively, respectively, while sugarcane bagasse type and particle size (within the range studied) did not influence kLa. Using large particles of untreated sugarcane bagasse, the loop-type bioreactors (CTA and SCA) exhibited higher mass transfer, compared to the BC reactor. At higher %S, SCA presented a higher kLa value (0.0448 s−1) than CTA, and the best operational conditions in terms of oxygen mass transfer were achieved for %S < 10.0 g L−1 and QAIR > 27.0 L min−1. These results demonstrated that pneumatic bioreactors can provide elevated oxygen transfer in the presence of vegetal biomass, making them an excellent option for use in three-phase systems for cellulolytic enzyme production by filamentous fungi.

Temporal Dynamics of In-Field Bioreactor Populations Reflect the Groundwater System and Respond Predictably to Perturbation.

PubMed

King, Andrew J; Preheim, Sarah P; Bailey, Kathryn L; Robeson, Michael S; Roy Chowdhury, Taniya; Crable, Bryan R; Hurt, Richard A; Mehlhorn, Tonia; Lowe, Kenneth A; Phelps, Tommy J; Palumbo, Anthony V; Brandt, Craig C; Brown, Steven D; Podar, Mircea; Zhang, Ping; Lancaster, W Andrew; Poole, Farris; Watson, David B; W Fields, Matthew; Chandonia, John-Marc; Alm, Eric J; Zhou, Jizhong; Adams, Michael W W; Hazen, Terry C; Arkin, Adam P; Elias, Dwayne A

2017-03-07

Temporal variability complicates testing the influences of environmental variability on microbial community structure and thus function. An in-field bioreactor system was developed to assess oxic versus anoxic manipulations on in situ groundwater communities. Each sample was sequenced (16S SSU rRNA genes, average 10,000 reads), and biogeochemical parameters are monitored by quantifying 53 metals, 12 organic acids, 14 anions, and 3 sugars. Changes in dissolved oxygen (DO), pH, and other variables were similar across bioreactors. Sequencing revealed a complex community that fluctuated in-step with the groundwater community and responded to DO. This also directly influenced the pH, and so the biotic impacts of DO and pH shifts are correlated. A null model demonstrated that bioreactor communities were driven in part not only by experimental conditions but also by stochastic variability and did not accurately capture alterations in diversity during perturbations. We identified two groups of abundant OTUs important to this system; one was abundant in high DO and pH and contained heterotrophs and oxidizers of iron, nitrite, and ammonium, whereas the other was abundant in low DO with the capability to reduce nitrate. In-field bioreactors are a powerful tool for capturing natural microbial community responses to alterations in geochemical factors beyond the bulk phase.

Temporal Dynamics of In-Field Bioreactor Populations Reflect the Groundwater System and Respond Predictably to Perturbation

DOE PAGES

King, Andrew J.; Preheim, Sarah P.; Bailey, Kathryn L.; ...

2017-01-23

Temporal variability complicates testing the influences of environmental variability on microbial community structure and thus function. An in-field bioreactor system was developed to assess oxic versus anoxic manipulations on in-situ groundwater communities. Each sample was sequenced (16S SSU rRNA genes, average 10,000 reads) and biogeochemical parameters monitored by quantifying 53 metals, 12 organic acids, 14 anions and 3 sugars. Changes in dissolved oxygen (DO), pH, and other variables were similar across bioreactors. Sequencing revealed a complex community that fluctuated in-step with the groundwater community, and responded to DO. This also directly influenced the pH and so the biotic impacts ofmore » DO and pH shifts are correlated. A null model demonstrated that bioreactor communities were driven in part by experimental conditions but also by stochastic variability and did not accurately capture alterations in diversity during perturbations. We identified two groups of abundant OTUs important to this system; one was abundant in high DO and pH and contained heterotrophs and oxidizers of iron, nitrite, and ammonium, whereas the other was abundant in low DO with the capability to reduce nitrate. In-field bioreactors are a powerful tool for capturing natural microbial community responses to alterations in geochemical factors beyond the bulk phase.« less

Temporal Dynamics of In-Field Bioreactor Populations Reflect the Groundwater System and Respond Predictably to Perturbation

DOE Office of Scientific and Technical Information (OSTI.GOV)

King, Andrew J.; Preheim, Sarah P.; Bailey, Kathryn L.

Temporal variability complicates testing the influences of environmental variability on microbial community structure and thus function. An in-field bioreactor system was developed to assess oxic versus anoxic manipulations on in-situ groundwater communities. Each sample was sequenced (16S SSU rRNA genes, average 10,000 reads) and biogeochemical parameters monitored by quantifying 53 metals, 12 organic acids, 14 anions and 3 sugars. Changes in dissolved oxygen (DO), pH, and other variables were similar across bioreactors. Sequencing revealed a complex community that fluctuated in-step with the groundwater community, and responded to DO. This also directly influenced the pH and so the biotic impacts ofmore » DO and pH shifts are correlated. A null model demonstrated that bioreactor communities were driven in part by experimental conditions but also by stochastic variability and did not accurately capture alterations in diversity during perturbations. We identified two groups of abundant OTUs important to this system; one was abundant in high DO and pH and contained heterotrophs and oxidizers of iron, nitrite, and ammonium, whereas the other was abundant in low DO with the capability to reduce nitrate. In-field bioreactors are a powerful tool for capturing natural microbial community responses to alterations in geochemical factors beyond the bulk phase.« less

JAXA-NASA Interoperability Demonstration for Application of DTN Under Simulated Rain Attenuation

NASA Technical Reports Server (NTRS)

Suzuki, Kiyoshisa; Inagawa, Shinichi; Lippincott, Jeff; Cecil, Andrew J.

2014-01-01

As is well known, K-band or higher band communications in space link segment often experience intermittent disruptions caused by heavy rainfall. In view of keeping data integrity and establishing autonomous operations under such situation, it is important to consider introducing a tolerance mechanism such as Delay/Disruption Tolerant Networking (DTN). The Consultative Committee for Space Data Systems (CCSDS) is studying DTN as part of the standardization activities for space data systems. As a contribution to CCSDS and a feasibility study for future utilization of DTN, Japan Aerospace Exploration Agency (JAXA) and National Aeronautics and Space Administration (NASA) conducted an interoperability demonstration for confirming its tolerance mechanism and capability of automatic operation using Data Relay Test Satellite (DRTS) space link and its ground terminals. Both parties used the Interplanetary Overlay Network (ION) open source software, including the Bundle Protocol, the Licklider Transmission Protocol, and Contact Graph Routing. This paper introduces the contents of the interoperability demonstration and its results.

Thin film bioreactors in space

NASA Technical Reports Server (NTRS)

Hughes-Fulford, M.; Scheld, H. W.

1989-01-01

Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization, and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers an opportunity to learn more about basic biological systems with one inmportant variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would make it possible to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

Define of internal recirculation coefficient for biological wastewater treatment in anoxic and aerobic bioreactors

NASA Astrophysics Data System (ADS)

Rossinskyi, Volodymyr

2018-02-01

The biological wastewater treatment technologies in anoxic and aerobic bioreactors with recycle of sludge mixture are used for the effective removal of organic compounds from wastewater. The change rate of sludge mixture recirculation between bioreactors leads to a change and redistribution of concentrations of organic compounds in sludge mixture in bioreactors and change hydrodynamic regimes in bioreactors. Determination of the coefficient of internal recirculation of sludge mixture between bioreactors is important for the choice of technological parameters of biological treatment (wastewater treatment duration in anoxic and aerobic bioreactors, flow capacity of recirculation pumps). Determination of the coefficient of internal recirculation of sludge mixture requires integrated consideration of hydrodynamic parameter (flow rate), kinetic parameter (rate of oxidation of organic compounds) and physical-chemical parameter of wastewater (concentration of organic compounds). The conducted numerical experiment from the proposed mathematical equations allowed to obtain analytical dependences of the coefficient of internal recirculation sludge mixture between bioreactors on the concentration of organic compounds in wastewater, the duration of wastewater treatment in bioreactors.

Large-scale Clinical-grade Retroviral Vector Production in a Fixed-Bed Bioreactor

PubMed Central

Wang, Xiuyan; Olszewska, Malgorzata; Qu, Jinrong; Wasielewska, Teresa; Bartido, Shirley; Hermetet, Gregory; Sadelain, Michel

2015-01-01

The successful genetic engineering of patient T cells with γ-retroviral vectors expressing chimeric antigen receptors or T-cell receptors for phase II clinical trials and beyond requires the large-scale manufacture of high-titer vector stocks. The production of retroviral vectors from stable packaging cell lines using roller bottles or 10- to 40-layer cell factories is limited by a narrow harvest window, labor intensity, open-system operations, and the requirement for significant incubator space. To circumvent these shortcomings, we optimized the production of vector stocks in a disposable fixed-bed bioreactor using good manufacturing practice–grade packaging cell lines. High-titer vector stocks were harvested over 10 days, representing a much broader harvest window than the 3-day harvest afforded by cell factories. For PG13 and 293Vec packaging cells, the average vector titer and the vector stocks’ yield in the bioreactor were higher by 3.2- to 7.3-fold, and 5.6- to 13.1-fold, respectively, than those obtained in cell factories. The vector production was 10.4 and 18.6 times more efficient than in cell factories for PG13 and 293Vec cells, respectively. Furthermore, the vectors produced from the fixed-bed bioreactors passed the release test assays for clinical applications. Therefore, a single vector lot derived from 293Vec is suitable to transduce up to 500 patients cell doses in the context of large clinical trials using chimeric antigen receptors or T-cell receptors. These findings demonstrate for the first time that a robust fixed-bed bioreactor process can be used to produce γ-retroviral vector stocks scalable up to the commercialization phase. PMID:25751502

Hydrofocusing Bioreactor Produces Anti-Cancer Alkaloids

NASA Technical Reports Server (NTRS)

Gonda, Steve R.; Valluri, Jagan V.

2011-01-01

A methodology for growing three-dimensional plant tissue models in a hydrodynamic focusing bioreactor (HFB) has been developed. The methodology is expected to be widely applicable, both on Earth and in outer space, as a means of growing plant cells and aggregates thereof under controlled conditions for diverse purposes, including research on effects of gravitation and other environmental factors upon plant growth and utilization of plant tissue cultures to produce drugs in quantities greater and at costs lower than those of conventional methodologies. The HFB was described in Hydro focus - ing Bioreactor for Three-Dimensional Cell Culture (MSC-22358), NASA Tech Briefs, Vol. 27, No. 3 (March 2003), page 66. To recapitulate: The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear liquid culture environment simultaneously with the herding of suspended cells and tissue assemblies and removal of unwanted air bubbles. The HFB includes a rotating cell-culture vessel with a centrally located sampling port and an internal rotating viscous spinner attached to a rotating base. The vessel and viscous spinner can be made to rotate at the same speed and direction or different speeds and directions to tailor the flow field and the associated hydrodynamic forces in the vessel in order to obtain low-shear suspension of cells and control of the locations of cells and air bubbles. For research and pharmaceutical-production applications, the HFB offers two major benefits: low shear stress, which promotes the assembly of cells into tissue-like three-dimensional constructs; and randomization of gravitational vectors relative to cells, which affects production of medicinal compounds. Presumably, apposition of plant cells in the absence of shear forces promotes cell-cell contacts, cell aggregation, and cell differentiation. Only gentle mixing is necessary for distributing nutrients and oxygen. It has been postulated that inasmuch as cells in the simulated

Design of a novel bioreactor and application in vascular tissue engineering

NASA Astrophysics Data System (ADS)

Zhang, Zhi-Xiong; Xi, Ting-Fei; Wang, Ying-Jun; Chen, Xiao-Song; Zhang, Jian; Wang, Chun-Ren; Gu, Yong-Quan; Chen, Liang; Li, Jian-Xin; Chen, Bing

2008-11-01

Endothelial cells (ECs) detachment under high shear stress at the early period of transplantation resulted in thrombosis and occlusion. To solve this problem, we developed a novel bioreactor. The bioreactor mimicked the formation of pulsatile flow in physiological conditions. Human umbilical vein ECs were seeded onto the lumen of living tissue conduits grown within dog peritoneal cavity. The shear stress generated by the bioreactor was increased step by step from 1.5 ± 0.8 dyn/cm 2 to 5.3 ± 2.4 dyn/cm 2, and was applied to ECs after static culture for 2 days. The results showed that completely confluent monolayer ECs were elongated, and were oriented parallel to the flow direction. The bioreactor could provide good environment for formation of endothelium. Stepwise increase shear stress could strengthen cell-cell and cell-extracellular matrix. The flow conditions of the bioreactor play a key role to determine the quality of the ECs lining.

NASA Environmentally Responsible Aviation's Highly-Loaded Front Block Compressor Demonstration

NASA Technical Reports Server (NTRS)

Celestina, Mark

2017-01-01

The ERA project was created in 2009 as part of NASAs Aeronautics Research Mission Directorates (ARMD) Integrated Systems Aviation Program (IASP). The purpose of the ERA project was to explore and document the feasibility, benefit, and technical risk of vehicles concepts and enabling technologies to reduce aviations impact on the environment. The metrics for this technology is given in Figure 1 with the N+2 metrics highlighted in green. It is anticipated that the United States air transportation system will continue to expand significantly over the next few decades thus adversely impacting the environment unless new technology is incorporated to simultaneously reduce nitrous oxides (NOx), noise and fuel consumption. In order to achieve the overall goals and meet the technology insertion challenges, these goals were divided into technical challenges that were to be achieved during the execution of the ERA project. Technical challenges were accomplished through test campaigns conducted by Integrated Technology Demonstration (ITDs). ERAs technical performance period ended in 2015.

Technology demonstration of starshade manufacturing for NASA's Exoplanet mission program

NASA Astrophysics Data System (ADS)

Kasdin, N. J.; Lisman, D.; Shaklan, S.; Thomson, M.; Cady, E.; Martin, S.; Marchen, L.; Vanderbei, R. J.; Macintosh, B.; Rudd, R. E.; Savransky, D.; Mikula, J.; Lynch, D.

2012-09-01

It is likely that the coming decade will see the development of a large visible light telescope with enabling technology for imaging exosolar Earthlike planets in the habitable zone of nearby stars. One such technology utilizes an external occulter, a satellite flying far from the telescope and employing a large screen, or starshade, to suppress the incoming starlight suffciently for detecting and characterizing exoplanets. This trades the added complexity of building the precisely shaped starshade and flying it in formation against simplifications in the telescope since extremely precise wavefront control is no longer necessary. In this paper we present the results of our project to design, manufacture, and measure a prototype occulter petal as part of NASA's first Technology Development for Exoplanet Missions program. We describe the mechanical design of the starshade and petal, the precision manufacturing tolerances, and the metrology approach. We demonstrate that the prototype petal meets the requirements and is consistent with a full-size occulter achieving better than 10-10 contrast.

Pyrosequence analysis of bacterial communities in aerobic bioreactors treating polycyclic aromatic hydrocarbon-contaminated soil

PubMed Central

Richardson, Stephen D.; Aitken, Michael D.

2011-01-01

Two aerobic, lab-scale, slurry-phase bioreactors were used to examine the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soil and the associated bacterial communities. The two bioreactors were operated under semi-continuous (draw-and-fill) conditions at a residence time of 35 days, but one was fed weekly and the other monthly. Most of the quantified PAHs, including high-molecular-weight compounds, were removed to a greater extent in the weekly-fed bioreactor, which achieved total PAH removal of 76%. Molecular analyses, including pyrosequencing of 16S rRNA genes, revealed significant shifts in the soil bacterial communities after introduction to the bioreactors and differences in the abundance and types of bacteria in each of the bioreactors. The weekly-fed bioreactor displayed a more stable bacterial community with gradual changes over time, whereas the monthly-fed bioreactor community was less consistent and may have been more strongly influenced by the influx of untreated soil during feeding. Phylogenetic groups containing known PAH-degrading bacteria previously identified through stable-isotope probing of the untreated soil were differentially affected by bioreactor conditions. Sequences from members of the Acidovorax and Sphingomonas genera, as well as the uncultivated ‘‘Pyrene Group 2’’ were abundant in the bioreactors. However, the relative abundances of sequences from the Pseudomonas, Sphingobium, and Pseudoxanthomonas genera, as well as from a group of unclassified anthracene degraders, were much lower in the bioreactors compared to the untreated soil. PMID:21369833

Small-scale, hydrogen-oxidizing-denitrifying bioreactor for treatment of nitrate-contaminated drinking water.

PubMed

Smith, Richard L; Buckwalter, Seanne P; Repert, Deborah A; Miller, Daniel N

2005-05-01

Nitrate removal by hydrogen-coupled denitrification was examined using flow-through, packed-bed bioreactors to develop a small-scale, cost effective system for treating nitrate-contaminated drinking-water supplies. Nitrate removal was accomplished using a Rhodocyclus sp., strain HOD 5, isolated from a sole-source drinking-water aquifer. The autotrophic capacity of the purple non-sulfur photosynthetic bacterium made it particularly adept for this purpose. Initial tests used a commercial bioreactor filled with glass beads and countercurrent, non-sterile flow of an autotrophic, air-saturated, growth medium and hydrogen gas. Complete removal of 2 mM nitrate was achieved for more than 300 days of operation at a 2-h retention time. A low-cost hydrogen generator/bioreactor system was then constructed from readily available materials as a water treatment approach using the Rhodocyclus strain. After initial tests with the growth medium, the constructed system was tested using nitrate-amended drinking water obtained from fractured granite and sandstone aquifers, with moderate and low TDS loads, respectively. Incomplete nitrate removal was evident in both water types, with high-nitrite concentrations in the bioreactor output, due to a pH increase, which inhibited nitrite reduction. This was rectified by including carbon dioxide in the hydrogen stream. Additionally, complete nitrate removal was accomplished with wastewater-impacted surface water, with a concurrent decrease in dissolved organic carbon. The results of this study using three chemically distinct water supplies demonstrate that hydrogen-coupled denitrification can serve as the basis for small-scale remediation and that pilot-scale testing might be the next logical step.

Small-scale, hydrogen-oxidizing-denitrifying bioreactor for treatment of nitrate-contaminated drinking water

USGS Publications Warehouse

Smith, R.L.; Buckwalter, S.P.; Repert, D.A.; Miller, D.N.

2005-01-01

Nitrate removal by hydrogen-coupled denitrification was examined using flow-through, packed-bed bioreactors to develop a small-scale, cost effective system for treating nitrate-contaminated drinking-water supplies. Nitrate removal was accomplished using a Rhodocyclus sp., strain HOD 5, isolated from a sole-source drinking-water aquifer. The autotrophic capacity of the purple non-sulfur photosynthetic bacterium made it particularly adept for this purpose. Initial tests used a commercial bioreactor filled with glass beads and countercurrent, non-sterile flow of an autotrophic, air-saturated, growth medium and hydrogen gas. Complete removal of 2 mM nitrate was achieved for more than 300 days of operation at a 2-h retention time. A low-cost hydrogen generator/bioreactor system was then constructed from readily available materials as a water treatment approach using the Rhodocyclus strain. After initial tests with the growth medium, the constructed system was tested using nitrate-amended drinking water obtained from fractured granite and sandstone aquifers, with moderate and low TDS loads, respectively. Incomplete nitrate removal was evident in both water types, with high-nitrite concentrations in the bioreactor output, due to a pH increase, which inhibited nitrite reduction. This was rectified by including carbon dioxide in the hydrogen stream. Additionally, complete nitrate removal was accomplished with wastewater-impacted surface water, with a concurrent decrease in dissolved organic carbon. The results of this study using three chemically distinct water supplies demonstrate that hydrogen-coupled denitrification can serve as the basis for small-scale remediation and that pilot-scale testing might be the next logical step.

DEMONSTRATION BULLETIN: SOIL WASHING SYSTEM - BIOTROL, INC.

EPA Science Inventory

The three component technologies of the BioTrol Soil Washing System (BSWS). Tested in the SITE demonstration were a Soil Washer (SW), and Aqueous Treatment System (ATS), and a Slurry Bio-Reactor (SBR). The Soil Washer operates on the principle that a significant fraction of the...

Hydrostatic pressure and shear stress affect endothelin-1 and nitric oxide release by endothelial cells in bioreactors.

PubMed

Vozzi, Federico; Bianchi, Francesca; Ahluwalia, Arti; Domenici, Claudio

2014-01-01

Abundant experimental evidence demonstrates that endothelial cells are sensitive to flow; however, the effect of fluid pressure or pressure gradients that are used to drive viscous flow is not well understood. There are two principal physical forces exerted on the blood vessel wall by the passage of intra-luminal blood: pressure and shear. To analyze the effects of pressure and shear independently, these two stresses were applied to cultured cells in two different types of bioreactors: a pressure-controlled bioreactor and a laminar flow bioreactor, in which controlled levels of pressure or shear stress, respectively, can be generated. Using these bioreactor systems, endothelin-1 (ET-1) and nitric oxide (NO) release from human umbilical vein endothelial cells were measured under various shear stress and pressure conditions. Compared to the controls, a decrease of ET-1 production by the cells cultured in both bioreactors was observed, whereas NO synthesis was up-regulated in cells under shear stress, but was not modulated by hydrostatic pressure. These results show that the two hemodynamic forces acting on blood vessels affect endothelial cell function in different ways, and that both should be considered when planning in vitro experiments in the presence of flow. Understanding the individual and synergic effects of the two forces could provide important insights into physiological and pathological processes involved in vascular remodeling and adaptation. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Concept designs for NASA's Solar Electric Propulsion Technology Demonstration Mission

NASA Technical Reports Server (NTRS)

Mcguire, Melissa L.; Hack, Kurt J.; Manzella, David H.; Herman, Daniel A.

2014-01-01

Multiple Solar Electric Propulsion Technology Demonstration Mission were developed to assess vehicle performance and estimated mission cost. Concepts ranged from a 10,000 kilogram spacecraft capable of delivering 4000 kilogram of payload to one of the Earth Moon Lagrange points in support of future human-crewed outposts to a 180 kilogram spacecraft capable of performing an asteroid rendezvous mission after launched to a geostationary transfer orbit as a secondary payload. Low-cost and maximum Delta-V capability variants of a spacecraft concept based on utilizing a secondary payload adapter as the primary bus structure were developed as were concepts designed to be co-manifested with another spacecraft on a single launch vehicle. Each of the Solar Electric Propulsion Technology Demonstration Mission concepts developed included an estimated spacecraft cost. These data suggest estimated spacecraft costs of $200 million - $300 million if 30 kilowatt-class solar arrays and the corresponding electric propulsion system currently under development are used as the basis for sizing the mission concept regardless of launch vehicle costs. The most affordable mission concept developed based on subscale variants of the advanced solar arrays and electric propulsion technology currently under development by the NASA Space Technology Mission Directorate has an estimated cost of $50M and could provide a Delta-V capability comparable to much larger spacecraft concepts.

A novel flow-perfusion bioreactor supports 3D dynamic cell culture.

PubMed

Sailon, Alexander M; Allori, Alexander C; Davidson, Edward H; Reformat, Derek D; Allen, Robert J; Warren, Stephen M

2009-01-01

Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm). A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm) scaffolds. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core. By day 8, static scaffolds had a periphery cell density of 67% +/- 5.0%, while in the core it was 0.3% +/- 0.3%. Flow-perfused scaffolds demonstrated peripheral cell density of 94% +/- 8.3% and core density of 76% +/- 3.1% at day 8. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.

LEACHATE NITROGEN CONCENTRATIONS AND BACTERIAL NUMBERS FROM TWO BIOREACTOR LANDFILLS

EPA Science Inventory

The U.S. EPA and Waste Management Inc. have entered into a cooperative research and development agreement (CRADA) to study landfills operated as bioreactors. Two different landfill bioreactor configurations are currently being tested at the Outer Loop landfill in Louisville, KY...

Breast Cancer Research at NASA

NASA Technical Reports Server (NTRS)

1998-01-01

Isolation of human mammary epithelial cells (HMEC) from breast cancer susceptible tissue; A: Duct element recovered from breast tissue digest. B: Outgrowth of cells from duct element in upper right corner cultured in a standard dish; most cells spontaneousely die during early cell divisions, but a few will establish long-term growth. C: Isolate of long-term frowth HMEC from outgrowth of duct element; cells shown soon after isolation and in early full-cell contact growth in culture in a dish. D: same long-term growth HMEC, but after 3 weeks in late full-cell contact growth in a continuous culture in a dish. Note attempts to reform duct elements but this in two demensions in a dish rather than in three dimensions in tissue. NASA's Marshall Space Flight Center (MSFC) is sponsoring research with Bioreactors, rotating wall vessels designed to grow tissue samples in space, to understand how breast cancer works. This ground-based work studies the growth and assembly of human mammary epithelial cell (HMEC) from breast cancer susceptible tissue. Radiation can make the cells cancerous, thus allowing better comparisons of healthy vs. tunorous tissue. Credit: Dr. Robert Richmond, NASA/Marshall Space Flight Center (MSFC).

Enhancement of matrix production and cell proliferation in human annulus cells under bioreactor culture.

PubMed

Yang, Xinlin; Wang, Daidong; Hao, Jianrong; Gong, Meiqing; Arlet, Vincent; Balian, Gary; Shen, Francis H; Li, Xudong Joshua

2011-06-01

Tissue engineering is a promising approach for treatment of disc degeneration. Herein, we evaluated effects of rotating bioreactor culture on the extracellular matrix production and proliferation of human annulus fibrosus (AF) cells. AF cells were embedded into alginate beads, and then cultured up to 3 weeks in a rotating wall vessel bioreactor or a static vessel. By real-time reverse transcription-polymerase chain reaction, expression of aggrecan, collagen type I and type II, and collagen prolyl 4-hydroxylase II was remarkably elevated, whereas expression of matrix metalloproteinase 3 and a disintegrin and metalloproteinase with thrombospondin motifs 5 was significantly decreased under bioreactor. Biochemical analysis revealed that the levels of the whole cell-associated proteoglycan and collagen were approximately five- and twofolds in rotating bioreactor, respectively, compared to those in static culture. Moreover, AF cell proliferation was augmented in rotating bioreactor. DNA contents were threefolds higher in rotating bioreactor than that in static culture. Expression of the proliferating cell nuclear antigen was robustly enhanced in rotating bioreactor as early as 1 week. Our findings suggested that rotating bioreactor culture would be an effective technique for expansion of human annulus cells for tissue engineering driven treatment of disc degeneration.

In Vitro Endothelialization of Biodegradable Vascular Grafts Via Endothelial Progenitor Cell Seeding and Maturation in a Tubular Perfusion System Bioreactor.

PubMed

Melchiorri, Anthony J; Bracaglia, Laura G; Kimerer, Lucas K; Hibino, Narutoshi; Fisher, John P

2016-07-01

A critical challenge to the success of biodegradable vascular grafts is the establishment of a healthy endothelium. To establish this monolayer of endothelial cells (ECs), a variety of techniques have been developed, including cell seeding. Vascular grafts may be seeded with relevant cell types and allowed to mature before implantation. Due to the low proliferative ability of adult ECs and issues with donor site morbidity, there has been increasing interest in using endothelial progenitor cells (EPCs) for vascular healing procedures. In this work, we combined the proliferative and differentiation capabilities of a commercial cell line of early EPCs with an established bioreactor system to support the maturation of cell-seeded vascular grafts. All components of the vascular graft and bioreactor setup are commercially available and allow for complete customization of the scaffold and culturing system. This bioreactor setup enables the control of flow through the graft, imparting fluid shear stress on EPCs and affecting cellular proliferation and differentiation. Grafts cultured with EPCs in the bioreactor system demonstrated greatly increased cell populations and neotissue formation compared with grafts seeded and cultured in a static system. Increased expression of markers for mature endothelial tissues were also observed in bioreactor-cultured EPC-seeded grafts. These findings suggest the distinct advantages of a customizable bioreactor setup for the proliferation and maturation of EPCs. Such a strategy may be beneficial for utilizing EPCs in vascular tissue engineering applications.

Fabrication, characterization, and in vitro evaluation of poly(lactic acid glycolic acid)/nano-hydroxyapatite composite microsphere-based scaffolds for bone tissue engineering in rotating bioreactors.

PubMed

Lv, Qing; Nair, Lakshmi; Laurencin, Cato T

2009-12-01

Dynamic flow culture bioreactor systems have been shown to enhance in vitro bone tissue formation by facilitating mass transfer and providing mechanical stimulation. Our laboratory has developed a biodegradable poly (lactic acid glycolic acid) (PLAGA) mixed scaffold consisting of lighter-than-water (LTW) and heavier-than-water (HTW) microspheres as potential matrices for engineering tissue using a high aspect ratio vessel (HARV) rotating bioreactor system. We have demonstrated enhanced osteoblast differentiation and mineralization on PLAGA scaffolds in the HARV rotating bioreactor system when compared with static culture. The objective of the present study is to improve the mechanical properties and bioactivity of polymeric scaffolds by designing LTW polymer/ceramic composite scaffolds suitable for dynamic culture using a HARV bioreactor. We employed a microsphere sintering method to fabricate three-dimensional PLAGA/nano-hydroxyapatite (n-HA) mixed scaffolds composed of LTW and HTW composite microspheres. The mechanical properties, pore size and porosity of the composite scaffolds were controlled by varying parameters, such as sintering temperature, sintering time, and PLAGA/n-HA ratio. The PLAGA/n-HA (4:1) scaffold sintered at 90 degrees C for 3 h demonstrated the highest mechanical properties and an appropriate pore structure for bone tissue engineering applications. Furthermore, evaluation human mesenchymal stem cells (HMSCs) response to PLAGA/n-HA scaffolds was performed. HMSCs on PLAGA/n-HA scaffolds demonstrated enhanced proliferation, differentiation, and mineralization when compared with those on PLAGA scaffolds. Therefore, PLAGA/n-HA mixed scaffolds are promising candidates for HARV bioreactor-based bone tissue engineering applications. Copyright 2008 Wiley Periodicals, Inc.

Characterization of TAP Ambr 250 disposable bioreactors, as a reliable scale-down model for biologics process development.

PubMed

Xu, Ping; Clark, Colleen; Ryder, Todd; Sparks, Colleen; Zhou, Jiping; Wang, Michelle; Russell, Reb; Scott, Charo

2017-03-01

Demands for development of biological therapies is rapidly increasing, as is the drive to reduce time to patient. In order to speed up development, the disposable Automated Microscale Bioreactor (Ambr 250) system is increasingly gaining interest due to its advantages, including highly automated control, high throughput capacity, and short turnaround time. Traditional early stage upstream process development conducted in 2 - 5 L bench-top bioreactors requires high foot-print, and running cost. The establishment of the Ambr 250 as a scale-down model leads to many benefits in process development. In this study, a comprehensive characterization of mass transfer coefficient (k L a) in the Ambr 250 was conducted to define optimal operational conditions. Scale-down approaches, including dimensionless volumetric flow rate (vvm), power per unit volume (P/V) and k L a have been evaluated using different cell lines. This study demonstrates that the Ambr 250 generated comparable profiles of cell growth and protein production, as seen at 5-L and 1000-L bioreactor scales, when using k L a as a scale-down parameter. In addition to mimicking processes at large scales, the suitability of the Ambr 250 as a tool for clone selection, which is traditionally conducted in bench-top bioreactors, was investigated. Data show that cell growth, productivity, metabolite profiles, and product qualities of material generated using the Ambr 250 were comparable to those from 5-L bioreactors. Therefore, Ambr 250 can be used for clone selection and process development as a replacement for traditional bench-top bioreactors minimizing resource utilization during the early stages of development in the biopharmaceutical industry. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:478-489, 2017. © 2017 American Institute of Chemical Engineers.

Protein Expression in Insect and Mammalian Cells Using Baculoviruses in Wave Bioreactors.

PubMed

Kadwell, Sue H; Overton, Laurie K

2016-01-01

Many types of disposable bioreactors for protein expression in insect and mammalian cells are now available. They differ in design, capacity, and sensor options, with many selections available for either rocking platform, orbitally shaken, pneumatically mixed, or stirred-tank bioreactors lined with an integral disposable bag (Shukla and Gottschalk, Trends Biotechnol 31(3):147-154, 2013). WAVE Bioreactors™ were among the first disposable systems to be developed (Singh, Cytotechnology 30:149-158, 1999). Since their commercialization in 1999, Wave Bioreactors have become routinely used in many laboratories due to their ease of operation, limited utility requirements, and protein expression levels comparability to traditional stirred-tank bioreactors. Wave Bioreactors are designed to use a presterilized Cellbag™, which is attached to a rocking platform and inflated with filtered air provided by the bioreactor unit. The Cellbag can be filled with medium and cells and maintained at a set temperature. The rocking motion, which is adjusted through angle and rock speed settings, provides mixing of oxygen (and CO2, which is used to control pH in mammalian cell cultures) from the headspace created in the inflated Cellbag with the cell culture medium and cells. This rocking motion can be adjusted to prevent cell shear damage. Dissolved oxygen and pH can be monitored during scale-up, and samples can be easily removed to monitor other parameters. Insect and mammalian cells grow very well in Wave Bioreactors (Shukla and Gottschalk, Trends Biotechnol 31(3):147-154, 2013). Combining Wave Bioreactor cell growth capabilities with recombinant baculoviruses engineered for insect or mammalian cell expression has proven to be a powerful tool for rapid production of a wide range of proteins.

Microgravity

NASA Image and Video Library

1996-01-01

Electronics control module for the NASA Bioreactor. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Microgravity

NASA Image and Video Library

1996-01-01

Interior view of the gas supply for the NASA Bioreactor. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Bioreactors for removing methyl bromide following contained fumigations

USGS Publications Warehouse

Miller, L.G.; Baesman, S.M.; Oremland, R.S.

2003-01-01

Use of methyl bromide (MeBr) as a quarantine, commodity, or structural fumigant is under scrutiny because its release to the atmosphere contributes to the depletion of stratospheric ozone. A closed-system bioreactor consisting of 0.5 L of a growing culture of a previously described bacterium, strain IMB-1, removed MeBr (> 110 ??mol L-1) from recirculating air. Strain IMB-1 grew slowly to high cell densities in the bioreactor using MeBr as its sole carbon and energy source. Bacterial oxidation of MeBr produced CO2 and hydrobromic acid (HBr), which required continuous neutralization with NaOH for the system to operate effectively. Strain IMB-1 was capable of sustained oxidation of large amounts of MeBr (170 mmol in 46 d). In an open-system bioreactor (10-L fermenter), strain IMB-1 oxidized a continuous supply of MeBr (220 ??mol L-1 in air). Growth was continuous, and 0.5 mol of MeBr was removed from the air supply in 14 d. The specific rate of MeBr oxidation was 7 ?? 10-16 mol cell-1 h-1. Bioreactors such as these can therefore be used to remove large quantities of contaminant MeBr, which opens the possibility of biodegradation as a practical means for its disposal.

3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor.

PubMed

Ball, Owen; Nguyen, Bao-Ngoc B; Placone, Jesse K; Fisher, John P

2016-12-01

There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state.

3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor

PubMed Central

Ball, Owen; Nguyen, Bao-Ngoc B.; Placone, Jesse K.; Fisher, John P.

2016-01-01

There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state. PMID:27272210

Architecture, component, and microbiome of biofilm involved in the fouling of membrane bioreactors.

PubMed

Inaba, Tomohiro; Hori, Tomoyuki; Aizawa, Hidenobu; Ogata, Atsushi; Habe, Hiroshi

2017-01-01

Biofilm formation on the filtration membrane and the subsequent clogging of membrane pores (called biofouling) is one of the most persistent problems in membrane bioreactors for wastewater treatment and reclamation. Here, we investigated the structure and microbiome of fouling-related biofilms in the membrane bioreactor using non-destructive confocal reflection microscopy and high-throughput Illumina sequencing of 16S rRNA genes. Direct confocal reflection microscopy indicated that the thin biofilms were formed and maintained regardless of the increasing transmembrane pressure, which is a common indicator of membrane fouling, at low organic-loading rates. Their solid components were primarily extracellular polysaccharides and microbial cells. In contrast, high organic-loading rates resulted in a rapid increase in the transmembrane pressure and the development of the thick biofilms mainly composed of extracellular lipids. High-throughput sequencing revealed that the biofilm microbiomes, including major and minor microorganisms, substantially changed in response to the organic-loading rates and biofilm development. These results demonstrated for the first time that the architectures, chemical components, and microbiomes of the biofilms on fouled membranes were tightly associated with one another and differed considerably depending on the organic-loading conditions in the membrane bioreactor, emphasizing the significance of alternative indicators other than the transmembrane pressure for membrane biofouling.

Microgravity

NASA Image and Video Library

1996-01-01

Laptop computer sits atop the Experiment Control Computer for a NASA Bioreactor. The flight crew can change operating conditions in the Bioreactor by using the graphical interface on the laptop. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

COMPUTER SIMULATOR (BEST) FOR DESIGNING SULFATE-REDUCING BACTERIA FIELD BIOREACTORS

EPA Science Inventory

BEST (bioreactor economics, size and time of operation) is a spreadsheet-based model that is used in conjunction with public domain software, PhreeqcI. BEST is used in the design process of sulfate-reducing bacteria (SRB) field bioreactors to passively treat acid mine drainage (A...

DESIGNING SULFATE-REDUCING BACTERIA FIELD BIOREACTORS USING THE BEST MODEL

EPA Science Inventory

BEST (bioreactor economics, size and time of operation) is a spreadsheet-based model that is used in conjunction with a public domain computer software package, PHREEQCI. BEST is intended to be used in the design process of sulfate-reducing bacteria (SRB)field bioreactors to pas...

NASA Social

NASA Image and Video Library

2011-05-18

Ed Mango, of the NASA Commercial Crew Office, speaks during a NASA Social, Friday, May 18, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

3D Magnetic Stem Cell Aggregation and Bioreactor Maturation for Cartilage Regeneration.

PubMed

Van de Walle, Aurore; Wilhelm, Claire; Luciani, Nathalie

2017-04-27

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach recently explored for the development of autologous replacements involves the differentiation of stem cells into chondrocytes. To initiate this chondrogenesis, a degree of compaction of the stem cells is required; hence, we demonstrated the feasibility of magnetically condensing cells, both within thick scaffolds and scaffold-free, using miniaturized magnetic field sources as cell attractors. This magnetic approach was also used to guide aggregate fusion and to build scaffold-free, organized, three-dimensional (3D) tissues several millimeters in size. In addition to having an enhanced size, the tissue formed by magnetic-driven fusion presented a significant increase in the expression of collagen II, and a similar trend was observed for aggrecan expression. As the native cartilage was subjected to forces that influenced its 3D structure, dynamic maturation was also performed. A bioreactor that provides mechanical stimuli was used to culture the magnetically seeded scaffolds over a 21-day period. Bioreactor maturation largely improved chondrogenesis into the cellularized scaffolds; the extracellular matrix obtained under these conditions was rich in collagen II and aggrecan. This work outlines the innovative potential of magnetic condensation of labeled stem cells and dynamic maturation in a bioreactor for improved chondrogenic differentiation, both scaffold-free and within polysaccharide scaffolds.

A Flow Perfusion Bioreactor System for Vocal Fold Tissue Engineering Applications

PubMed Central

Heris, Hossein K.; Thomson, Scott L.; Taher, Rani; Kazemirad, Siavash; Sheibani, Sara; Li-Jessen, Nicole Y.K.; Vali, Hojatollah; Mongeau, Luc

2016-01-01

The human vocal folds (VFs) undergo complex biomechanical stimulation during phonation. The aim of the present study was to develop and validate a phono-mimetic VF flow perfusion bioreactor, which mimics the mechanical microenvironment of the human VFs in vitro. The bioreactor uses airflow-induced self-oscillations, which have been shown to produce mechanical loading and contact forces that are representative of human phonation. The bioreactor consisted of two synthetic VF replicas within a silicone body. A cell-scaffold mixture (CSM) consisting of human VF fibroblasts, hyaluronic acid, gelatin, and a polyethylene glycol cross-linker was injected into cavities within the replicas. Cell culture medium (CCM) was perfused through the scaffold by using a customized secondary flow loop. After the injection, the bioreactor was operated with no stimulation over a 3-day period to allow for cell adaptation. Phonation was subsequently induced by using a variable speed centrifugal blower for 2 h each day over a period of 4 days. A similar bioreactor without biomechanical stimulation was used as the nonphonatory control. The CSM was harvested from both VF replicas 7 days after the injection. The results confirmed that the phono-mimetic bioreactor supports cell viability and extracellular matrix proteins synthesis, as expected. Many scaffold materials were found to degrade because of challenges from phonation-induced biomechanical stimulation as well as due to biochemical reactions with the CCM. The bioreactor concept enables future investigations of the effects of different phonatory characteristics, that is, voice regimes, on the behavior of the human VF cells. It will also help study the long-term functional outcomes of the VF-specific biomaterials before animal and clinical studies. PMID:27537192

A Flow Perfusion Bioreactor System for Vocal Fold Tissue Engineering Applications.

PubMed

Latifi, Neda; Heris, Hossein K; Thomson, Scott L; Taher, Rani; Kazemirad, Siavash; Sheibani, Sara; Li-Jessen, Nicole Y K; Vali, Hojatollah; Mongeau, Luc

2016-09-01

The human vocal folds (VFs) undergo complex biomechanical stimulation during phonation. The aim of the present study was to develop and validate a phono-mimetic VF flow perfusion bioreactor, which mimics the mechanical microenvironment of the human VFs in vitro. The bioreactor uses airflow-induced self-oscillations, which have been shown to produce mechanical loading and contact forces that are representative of human phonation. The bioreactor consisted of two synthetic VF replicas within a silicone body. A cell-scaffold mixture (CSM) consisting of human VF fibroblasts, hyaluronic acid, gelatin, and a polyethylene glycol cross-linker was injected into cavities within the replicas. Cell culture medium (CCM) was perfused through the scaffold by using a customized secondary flow loop. After the injection, the bioreactor was operated with no stimulation over a 3-day period to allow for cell adaptation. Phonation was subsequently induced by using a variable speed centrifugal blower for 2 h each day over a period of 4 days. A similar bioreactor without biomechanical stimulation was used as the nonphonatory control. The CSM was harvested from both VF replicas 7 days after the injection. The results confirmed that the phono-mimetic bioreactor supports cell viability and extracellular matrix proteins synthesis, as expected. Many scaffold materials were found to degrade because of challenges from phonation-induced biomechanical stimulation as well as due to biochemical reactions with the CCM. The bioreactor concept enables future investigations of the effects of different phonatory characteristics, that is, voice regimes, on the behavior of the human VF cells. It will also help study the long-term functional outcomes of the VF-specific biomaterials before animal and clinical studies.

Design and evaluation of a bioreactor with application to forensic burial environments.

PubMed

Dunphy, Melissa A; Weisensee, Katherine E; Mikhailova, Elena A; Harman, Melinda K

2015-12-01

Existing forensic taphonomic methods lack specificity in estimating the postmortem interval (PMI) in the period following active decomposition. New methods, such as the use of citrate concentration in bone, are currently being considered; however, determining the applicability of these methods in differing environmental contexts is challenging. This research aims to design a forensic bioreactor that can account for environmental factors known to impact decomposition, specifically temperature, moisture, physical damage from animals, burial depth, soil pH, and organic matter content. These forensically relevant environmental variables were characterized in a soil science context. The resulting metrics were soil temperature regime, soil moisture regime, slope, texture, soil horizon, cation exchange capacity, soil pH, and organic matter content. Bioreactor chambers were constructed using sterilized thin-walled polystyrene boxes housed in calibrated temperature units. Gravesoil was represented using mineral soil (Ultisols), and organic soil proxy for Histosols, horticulture mix. Gravesoil depth was determined using mineral soil horizons A and Bt2 to simulate surface scatter and shallow grave burial respectively. A total of fourteen different environmental conditions were created and controlled successfully over a 90-day experiment. These results demonstrate successful implementation and control of forensic bioreactor simulating precise environments in a single research location, rather than site-specific testing occurring in different geographic regions. Bone sections were grossly assessed for weathering characteristics, which revealed notable differences related to exposure to different temperature regimes and soil types. Over the short 90-day duration of this experiment, changes in weathering characteristics were more evident across the different temperature regimes rather than the soil types. Using this methodology, bioreactor systems can be created to replicate many

Effects of aeration frequency on leachate quality and waste in simulated hybrid bioreactor landfills.

PubMed

Ko, Jae Hac; Ma, Zeyu; Jin, Xiao; Xu, Qiyong

2016-12-01

Research has been conducted to investigate the effects of daily aeration frequency on leachate quality and waste settlement in simulated hybrid landfill bioreactors. Four laboratory-scale reactors were constructed and operated for about 10 months to simulate different bioreactor operations, including one anaerobic bioreactor and three hybrid bioreactors with different aeration frequencies (one, two, and four times per day). Chemical oxygen demand (COD) and biochemical oxygen demand (BOD 5 ) reduced more than 96% of the initial concentrations in all aerated bioreactors. The differences of COD and BOD 5 reductions among tested aeration frequencies were relatively small. For ammonia nitrogen, the higher aeration frequency (two or four times per day) resulted in the quicker reduction. Overall, the concentrations of heavy metals (Cr, Co, Cu, Mn, Ni, and Zn) decreased over time except Cd and Pb. The reduction of redox-sensitive metal concentrations (Mn, Co, Ni, and Cu) was greater in aerated bioreactors than in anaerobic bioreactor. Settlement of municipal solid waste (MSW) was enhanced with higher frequency of aeration events (four times per day). In recent years, hybird bioreactor landfill technology has gained a lot of attention. Appropriate aeration rate is crucial for hybrid bioreactor operation, but few studies have been done and different results were obtained. Research was conducted to investigate the effects of daily aeration frequency on leachate quality and waste settlement. Results indicated that aeration can effectively accelerate waste stabilization and remove organic carbon concentration and total nitrogen in the leachate.

Membrane bioreactors for the removal of anionic micropollutants from drinking water.

PubMed

Crespo, João G; Velizarov, Svetlozar; Reis, Maria A

2004-10-01

Biological treatment processes allow for the effective elimination of anionic micropollutants from drinking water. However, special technologies have to be implemented to eliminate the target pollutants without changing water quality, either by adding new pollutants or removing essential water components. Some innovative technologies that combine the use of membranes with the biological degradation of ionic micropollutants in order to minimize the secondary contamination of treated water include pressure-driven membrane bioreactors, gas-transfer membrane bioreactors and ion exchange membrane bioreactors.

Development and Characterization of a Parallelizable Perfusion Bioreactor for 3D Cell Culture.

PubMed

Egger, Dominik; Fischer, Monica; Clementi, Andreas; Ribitsch, Volker; Hansmann, Jan; Kasper, Cornelia

2017-05-25

The three dimensional (3D) cultivation of stem cells in dynamic bioreactor systems is essential in the context of regenerative medicine. Still, there is a lack of bioreactor systems that allow the cultivation of multiple independent samples under different conditions while ensuring comprehensive control over the mechanical environment. Therefore, we developed a miniaturized, parallelizable perfusion bioreactor system with two different bioreactor chambers. Pressure sensors were also implemented to determine the permeability of biomaterials which allows us to approximate the shear stress conditions. To characterize the flow velocity and shear stress profile of a porous scaffold in both bioreactor chambers, a computational fluid dynamics analysis was performed. Furthermore, the mixing behavior was characterized by acquisition of the residence time distributions. Finally, the effects of the different flow and shear stress profiles of the bioreactor chambers on osteogenic differentiation of human mesenchymal stem cells were evaluated in a proof of concept study. In conclusion, the data from computational fluid dynamics and shear stress calculations were found to be predictable for relative comparison of the bioreactor geometries, but not for final determination of the optimal flow rate. However, we suggest that the system is beneficial for parallel dynamic cultivation of multiple samples for 3D cell culture processes.

Development and Characterization of a Parallelizable Perfusion Bioreactor for 3D Cell Culture

PubMed Central

Egger, Dominik; Fischer, Monica; Clementi, Andreas; Ribitsch, Volker; Hansmann, Jan; Kasper, Cornelia

2017-01-01

The three dimensional (3D) cultivation of stem cells in dynamic bioreactor systems is essential in the context of regenerative medicine. Still, there is a lack of bioreactor systems that allow the cultivation of multiple independent samples under different conditions while ensuring comprehensive control over the mechanical environment. Therefore, we developed a miniaturized, parallelizable perfusion bioreactor system with two different bioreactor chambers. Pressure sensors were also implemented to determine the permeability of biomaterials which allows us to approximate the shear stress conditions. To characterize the flow velocity and shear stress profile of a porous scaffold in both bioreactor chambers, a computational fluid dynamics analysis was performed. Furthermore, the mixing behavior was characterized by acquisition of the residence time distributions. Finally, the effects of the different flow and shear stress profiles of the bioreactor chambers on osteogenic differentiation of human mesenchymal stem cells were evaluated in a proof of concept study. In conclusion, the data from computational fluid dynamics and shear stress calculations were found to be predictable for relative comparison of the bioreactor geometries, but not for final determination of the optimal flow rate. However, we suggest that the system is beneficial for parallel dynamic cultivation of multiple samples for 3D cell culture processes. PMID:28952530

Strategies for Enhancing the Accumulation and Retention of Extracellular Matrix in Tissue-Engineered Cartilage Cultured in Bioreactors

PubMed Central

Shahin, Kifah; Doran, Pauline M.

2011-01-01

Production of tissue-engineered cartilage involves the synthesis and accumulation of key constituents such as glycosaminoglycan (GAG) and collagen type II to form insoluble extracellular matrix (ECM). During cartilage culture, macromolecular components are released from nascent tissues into the medium, representing a significant waste of biosynthetic resources. This work was aimed at developing strategies for improving ECM retention in cartilage constructs and thus the quality of engineered tissues produced in bioreactors. Human chondrocytes seeded into polyglycolic acid (PGA) scaffolds were cultured in perfusion bioreactors for up to 5 weeks. Analysis of the size and integrity of proteoglycans in the constructs and medium showed that full-sized aggrecan was being stripped from the tissues without proteolytic degradation. Application of low (0.075 mL min−1) and gradually increasing (0.075–0.2 mL min−1) medium flow rates in the bioreactor resulted in the generation of larger constructs, a 4.0–4.4-fold increase in the percentage of GAG retained in the ECM, and a 4.8–5.2-fold increase in GAG concentration in the tissues compared with operation at 0.2 mL min−1. GAG retention was also improved by pre-culturing seeded scaffolds in flasks for 5 days prior to bioreactor culture. In contrast, GAG retention in PGA scaffolds infused with alginate hydrogel did not vary significantly with medium flow rate or pre-culture treatment. This work demonstrates that substantial improvements in cartilage quality can be achieved using scaffold and bioreactor culture strategies that specifically target and improve ECM retention. PMID:21858004

Expansion of Human Mesenchymal Stem Cells in a Microcarrier Bioreactor.

PubMed

Tsai, Ang-Chen; Ma, Teng

2016-01-01

Human mesenchymal stem cells (hMSCs) are considered as a primary candidate in cell therapy owing to their self-renewability, high differentiation capabilities, and secretions of trophic factors. In clinical application, a large quantity of therapeutically competent hMSCs is required that cannot be produced in conventional petri dish culture. Bioreactors are scalable and have the capacity to meet the production demand. Microcarrier suspension culture in stirred-tank bioreactors is the most widely used method to expand anchorage dependent cells in a large scale. Stirred-tank bioreactors have the potential to scale up and microcarriers provide the high surface-volume ratio. As a result, a spinner flask bioreactor with microcarriers has been commonly used in large scale expansion of adherent cells. This chapter describes a detailed culture protocol for hMSC expansion in a 125 mL spinner flask using microcarriers, Cytodex I, and a procedure for cell seeding, expansion, metabolic sampling, and quantification and visualization using microculture tetrazolium (MTT) reagent.

NASA Social

NASA Image and Video Library

2012-05-19

A NASA Social follower holds up a mobile device as NASA Administrator Charles Bolden, left, and Kennedy Space Center director Robert Cabana appear at the NASA Social event, Friday morning, May 19, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

[Research progress of in vivo bioreactor as vascularization strategies in bone tissue engineering].

PubMed

Zhang, Haifeng; Han, Dong

2014-09-01

To review the application and research progress of in vivo bioreactor as vascularization strategies in bone tissue engineering. The original articles about in vivo bioreactor that can enhance vascularization of tissue engineered bone were extensively reviewed and analyzed. The in vivo bioreactor can be created by periosteum, muscle, muscularis membrane, and fascia flap as well as biomaterials. Using in vivo bioreactor can effectively promote the establishment of a microcirculation in the tissue engineered bones, especially for large bone defects. However, main correlative researches, currently, are focused on animal experiments, more clinical trials will be carried out in the future. With the rapid development of related technologies of bone tissue engineering, the use of in vivo bioreactor will to a large extent solve the bottleneck limitations and has the potential values for clinical application.

Use of a Packed-Column Bioreactor for Isolation of Diverse Protease-Producing Bacteria from Antarctic Soil

PubMed Central

Wery, Nathalie; Gerike, Ursula; Sharman, Ajay; Chaudhuri, Julian B.; Hough, David W.; Danson, Michael J.

2003-01-01

Seventy-five aerobic heterotrophs have been isolated from a packed-column bioreactor inoculated with soil from Antarctica. The column was maintained at 10°C and continuously fed with a casein-containing medium to enrich protease producers. Twenty-eight isolates were selected for further characterization on the basis of morphology and production of clearing zones on skim milk plates. Phenotypic tests indicated that the strains were mainly psychrotrophs and presented a high morphological and metabolical diversity. The extracellular protease activities tested were optimal at neutral pH and between 30 and 45°C. 16S ribosomal DNA sequence analyses showed that the bioreactor was colonized by a wide variety of taxons, belonging to various bacterial divisions: α-, β-, and γ-Proteobacteria; the Flexibacter-Cytophaga-Bacteroides group; and high G+C gram-positive bacteria and low G+C gram-positive bacteria. Some strains represent candidates for new species of the genera Chryseobacterium and Massilia. This diversity demonstrates that the bioreactor is an efficient enrichment tool compared to traditional isolation strategies. PMID:12620829

Large-scale progenitor cell expansion for multiple donors in a monitored hollow fibre bioreactor.

PubMed

Lambrechts, Toon; Papantoniou, Ioannis; Rice, Brent; Schrooten, Jan; Luyten, Frank P; Aerts, Jean-Marie

2016-09-01

With the increasing scale in stem cell production, a robust and controlled cell expansion process becomes essential for the clinical application of cell-based therapies. The objective of this work was the assessment of a hollow fiber bioreactor (Quantum Cell Expansion System from Terumo BCT) as a cell production unit for the clinical-scale production of human periosteum derived stem cells (hPDCs). We aimed to demonstrate comparability of bioreactor production to standard culture flask production based on a product characterization in line with the International Society of Cell Therapy in vitro benchmarks and supplemented with a compelling quantitative in vivo bone-forming potency assay. Multiple process read-outs were implemented to track process performance and deal with donor-to-donor-related variation in nutrient needs and harvest timing. The data show that the hollow fiber bioreactor is capable of robustly expanding autologous hPDCs on a clinical scale (yield between 316 million and 444 million cells starting from 20 million after ± 8 days of culture) while maintaining their in vitro quality attributes compared with the standard flask-based culture. The in vivo bone-forming assay on average resulted in 10.3 ± 3.7% and 11.0 ± 3.8% newly formed bone for the bioreactor and standard culture flask respectively. The analysis showed that the Quantum system provides a reproducible cell expansion process in terms of yields and culture conditions for multiple donors. Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Fish as bioreactors: transgene expression of human coagulation factor VII in fish embryos.

PubMed

Hwang, Gyulin; Müller, Ferenc; Rahman, M Aziz; Williams, Darren W; Murdock, Paul J; Pasi, K John; Goldspink, Geoffrey; Farahmand, Hamid; Maclean, Norman

2004-01-01

A plasmid containing human coagulation factor VII (hFVII) complementary DNA regulated by a cytomegalovirus promoter was microinjected into fertilized eggs of zebrafish, African catfish, and tilapia. The active form of hFVll was detected in the fish embryos by various assays. This positive expression of human therapeutic protein in fish embryos demonstrates the possibility of exploitation of transgenic fish as bioreactors.

NDARC - NASA Design and Analysis of Rotorcraft Validation and Demonstration

NASA Technical Reports Server (NTRS)

Johnson, Wayne

2010-01-01

Validation and demonstration results from the development of the conceptual design tool NDARC (NASA Design and Analysis of Rotorcraft) are presented. The principal tasks of NDARC are to design a rotorcraft to satisfy specified design conditions and missions, and then analyze the performance of the aircraft for a set of off-design missions and point operating conditions. The aircraft chosen as NDARC development test cases are the UH-60A single main-rotor and tail-rotor helicopter, the CH-47D tandem helicopter, the XH-59A coaxial lift-offset helicopter, and the XV-15 tiltrotor. These aircraft were selected because flight performance data, a weight statement, detailed geometry information, and a correlated comprehensive analysis model are available for each. Validation consists of developing the NDARC models for these aircraft by using geometry and weight information, airframe wind tunnel test data, engine decks, rotor performance tests, and comprehensive analysis results; and then comparing the NDARC results for aircraft and component performance with flight test data. Based on the calibrated models, the capability of the code to size rotorcraft is explored.

Biotechnology

NASA Image and Video Library

2001-05-15

This prostate cancer construct was grown during NASA-sponsored bioreactor studies on Earth. Cells are attached to a biodegradable plastic lattice that gives them a head start in growth. Prostate tumor cells are to be grown in a NASA-sponsored Bioreactor experiment aboard the STS-107 Research-1 mission in 2002. Dr. Leland Chung of the University of Virginia is the principal investigator. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Credit: NASA and the University of Virginia.

Seasonal Patterns in Microbial Community Composition in Denitrifying Bioreactors Treating Subsurface Agricultural Drainage.

PubMed

Porter, Matthew D; Andrus, J Malia; Bartolerio, Nicholas A; Rodriguez, Luis F; Zhang, Yuanhui; Zilles, Julie L; Kent, Angela D

2015-10-01

Denitrifying bioreactors, consisting of water flow control structures and a woodchip-filled trench, are a promising approach for removing nitrate from agricultural subsurface or tile drainage systems. To better understand the seasonal dynamics and the ecological drivers of the microbial communities responsible for denitrification in these bioreactors, we employed microbial community "fingerprinting" techniques in a time-series examination of three denitrifying bioreactors over 2 years, looking at bacteria, fungi, and the denitrifier functional group responsible for the final step of complete denitrification. Our analysis revealed that microbial community composition responds to depth and seasonal variation in moisture content and inundation of the bioreactor media, as well as temperature. Using a geostatistical analysis approach, we observed recurring temporal patterns in bacterial and denitrifying bacterial community composition in these bioreactors, consistent with annual cycling. The fungal communities were more stable, having longer temporal autocorrelations, and did not show significant annual cycling. These results suggest a recurring seasonal cycle in the denitrifying bioreactor microbial community, likely due to seasonal variation in moisture content.

The Fluid Mechanics of a Wavy-Wall Bioreactor

NASA Astrophysics Data System (ADS)

Sucosky, Philippe; Bilgen, Bahar; Aleem, Alexander; Neitzel, Paul; Barabino, Gilda

2004-11-01

Bioreactors are devices used for the production of mammalian tissue in vitro. Although mixing has been shown to stimulate the growth of cartilage constructs, high shear-stress levels can damage the cells. In order to enhance mixing while minimizing shear, a wavy-wall bioreactor (WWB) featuring a sinusoidal internal profile has been designed. The turbulent hydrodynamic environment produced in this device is investigated experimentally using particle-image velocimetry. A model bioreactor made of acrylic and filled with an index-matching solution of zinc iodide is used to compensate for the refraction of light at the walls. The flow observed in different planes is shown to be periodic, spatially dependent, and dominated by mean-shear rather than Reynolds stresses in the vicinity of constructs. Finally, a comparison between the mean-shear stresses obtained in the WWB and in a standard spinner flask reveals similar stress levels near the construct walls.

Biodegradation of paint stripper solvents in a modified gas lift loop bioreactor.

PubMed

Vanderberg-Twary, L; Steenhoudt, K; Travis, B J; Hanners, J L; Foreman, T M; Brainard, J R

1997-07-05

Paint stripping wastes generated during the decontamination and decommissioning of former nuclear facilities contain paint stripping organics (dichloromethane, 2-propanol, and methanol) and bulk materials containing paint pigments. It is desirable to degrade the organic residues as part of an integrated chemical-biological treatment system. We have developed a modified gas lift loop bioreactor employing a defined consortium of Rhodococcus rhodochrous strain OFS and Hyphomicrobium sp. DM-2 that degrades paint stripper organics. Mass transfer coefficients and kinetic constants for biodegradation in the system were determined. It was found that transfer of organic substrates from surrogate waste into the air and further into the liquid medium in the bioreactor were rapid processes, occurring within minutes. Monod kinetics was employed to model the biodegradation of paint stripping organics. Analysis of the bioreactor process was accomplished with BIOLAB, a mathematical code that simulates coupled mass transfer and biodegradation processes. This code was used to fit experimental data to Monod kinetics and to determine kinetic parameters. The BIOLAB code was also employed to compare activities in the bioreactor of individual microbial cultures to the activities of combined cultures in the bioreactor. This code is of benefit for further optimization and scale-up of the bioreactor for treatment of paint stripping and other volatile organic wastes in bulk materials.

The Role of Bioreactors in Ligament and Tendon Tissue Engineering.

PubMed

Mace, James; Wheelton, Andy; Khan, Wasim S; Anand, Sanj

2016-01-01

Bioreactors are pivotal to the emerging field of tissue engineering. The formation of neotissue from pluripotent cell lineages potentially offers a source of tissue for clinical use without the significant donor site morbidity associated with many contemporary surgical reconstructive procedures. Modern bioreactor design is becoming increasingly complex to provide a both an expandable source of readily available pluripotent cells and to facilitate their controlled differentiation into a clinically applicable ligament or tendon like neotissue. This review presents the need for such a method, challenges in the processes to engineer neotissue and the current designs and results of modern bioreactors in the pursuit of engineered tendon and ligament.

New generation NMR bioreactor coupled with high-resolution NMR spectroscopy leads to novel discoveries in Moorella thermoaceticum metabolic profiles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xue, Junfeng; Isern, Nancy G.; Ewing, R James

An in-situ nuclear magnetic resonance (NMR) bioreactor was developed and employed to monitor microbial metabolism under batch-growth conditions in real time. We selected Moorella thermoacetica ATCC 49707 as a test case. M. thermoacetica (formerly Clostridium thermoaceticum) is a strictly anaerobic, thermophilic, acetogenic, gram-positive bacterium with potential for industrial production of chemicals. The metabolic profiles of M. thermoacetica were characterized during growth in batch mode on xylose (a component of lignocellulosic biomass) using the new generation NMR bioreactor in combination with high-resolution, high sensitivity NMR (HR-NMR) spectroscopy. In-situ NMR measurements were performed using water-suppressed H-1 NMR spectroscopy at an NMR frequencymore » of 500 MHz, and aliquots of the bioreactor contents were taken for 600 MHz HR-NMR spectroscopy at specific intervals to confirm metabolite identifications and expand metabolite coverage. M. thermoacetica demonstrated the metabolic potential to produce formate, ethanol and methanol from xylose, in addition to its known capability of producing acetic acid. Real-time monitoring of bioreactor conditions showed a temporary pH decrease, with a concomitant increase in formic acid during exponential growth. Fermentation experiments performed outside of the magnet showed that the strong magnetic field employed for NMR detection did not significantly affect cell metabolism. Use of the in-situ NMR bioreactor facilitated monitoring of the fermentation process in real time, enabling identification of intermediate and end-point metabolites and their correlation with pH and biomass produced during culture growth. Real-time monitoring of culture metabolism using the NMR bioreactor in combination with the HR-NMR spectroscopy will allow optimization of the metabolism of microorganisms producing valuable bioproducts.« less

A novel perfused rotary bioreactor for cardiomyogenesis of embryonic stem cells.

PubMed

Teo, Ailing; Mantalaris, Athanasios; Song, Kedong; Lim, Mayasari

2014-05-01

Developments in bioprocessing technology play an important role for overcoming challenges in cardiac tissue engineering. To this end, our laboratory has developed a novel rotary perfused bioreactor for supporting three-dimensional cardiac tissue engineering. The dynamic culture environments provided by our novel perfused rotary bioreactor and/or the high-aspect rotating vessel produced constructs with higher viability and significantly higher cell numbers (up to 4 × 10(5) cells/bead) than static tissue culture flasks. Furthermore, cells in the perfused rotary bioreactor showed earlier gene expressions of cardiac troponin-T, α- and β-myosin heavy chains with higher percentages of cardiac troponin-I-positive cells and better uniformity of sacromeric α-actinin expression. A dynamic and perfused environment, as provided by this bioreactor, provides a superior culture performance in cardiac differentiation for embryonic stem cells particularly for larger 3D constructs.

Numerical simulation of microcarrier motion in a rotating wall vessel bioreactor.

PubMed

Ju, Zhi-Hao; Liu, Tian-Qing; Ma, Xue-Hu; Cui, Zhan-Feng

2006-06-01

To analyze the forces of rotational wall vessel (RWV) bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. The motion of the microcarrier in the rotating wall vessel (RWV) bioreactor with both the inner and outer cylinders rotating was modeled by numerical simulation. The continuous trajectory of microcarrier particles, including the possible collision with the wall was obtained. An expression between the minimum rotational speed difference of the inner and outer cylinders and the microcarrier particle or aggregate radius could avoid collisions with either wall. The range of microcarrier radius or tissue size, which could be safely cultured in the RWV bioreactor, in terms of shear stress level, was determined. The model works well in describing the trajectory of a heavier microcarrier particle in rotating wall vessel.

NASA Social

NASA Image and Video Library

2012-05-19

NASA Administrator Charles Bolden, left, and Kennedy Space Center director Robert Cabana appear at the NASA Social event, Friday morning, May 19, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

Bioreactor and methods for producing synchronous cells

NASA Technical Reports Server (NTRS)

Helmstetter, Charles E. (Inventor); Thornton, Maureen (Inventor); Gonda, Steve (Inventor)

2005-01-01

Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

Accelerated and Improved Differentiation of Retinal Organoids from Pluripotent Stem Cells in Rotating-Wall Vessel Bioreactors.

PubMed

DiStefano, Tyler; Chen, Holly Yu; Panebianco, Christopher; Kaya, Koray Dogan; Brooks, Matthew J; Gieser, Linn; Morgan, Nicole Y; Pohida, Tom; Swaroop, Anand

2018-01-09

Pluripotent stem cells can be differentiated into 3D retinal organoids, with major cell types self-patterning into a polarized, laminated architecture. In static cultures, organoid development may be hindered by limitations in diffusion of oxygen and nutrients. Herein, we report a bioprocess using rotating-wall vessel (RWV) bioreactors to culture retinal organoids derived from mouse pluripotent stem cells. Organoids in RWV demonstrate enhanced proliferation, with well-defined morphology and improved differentiation of neurons including ganglion cells and S-cone photoreceptors. Furthermore, RWV organoids at day 25 (D25) reveal similar maturation and transcriptome profile as those at D32 in static culture, closely recapitulating spatiotemporal development of postnatal day 6 mouse retina in vivo. Interestingly, however, retinal organoids do not differentiate further under any in vitro condition tested here, suggesting additional requirements for functional maturation. Our studies demonstrate that bioreactors can accelerate and improve organoid growth and differentiation for modeling retinal disease and evaluation of therapies. Published by Elsevier Inc.

Nano-ceramic composite scaffolds for bioreactor-based bone engineering.

PubMed

Lv, Qing; Deng, Meng; Ulery, Bret D; Nair, Lakshmi S; Laurencin, Cato T

2013-08-01

Composites of biodegradable polymers and bioactive ceramics are candidates for tissue-engineered scaffolds that closely match the properties of bone. We previously developed a porous, three-dimensional poly (D,L-lactide-co-glycolide) (PLAGA)/nanohydroxyapatite (n-HA) scaffold as a potential bone tissue engineering matrix suitable for high-aspect ratio vessel (HARV) bioreactor applications. However, the physical and cellular properties of this scaffold are unknown. The present study aims to evaluate the effect of n-HA in modulating PLAGA scaffold properties and human mesenchymal stem cell (HMSC) responses in a HARV bioreactor. By comparing PLAGA/n-HA and PLAGA scaffolds, we asked whether incorporation of n-HA (1) accelerates scaffold degradation and compromises mechanical integrity; (2) promotes HMSC proliferation and differentiation; and (3) enhances HMSC mineralization when cultured in HARV bioreactors. PLAGA/n-HA scaffolds (total number = 48) were loaded into HARV bioreactors for 6 weeks and monitored for mass, molecular weight, mechanical, and morphological changes. HMSCs were seeded on PLAGA/n-HA scaffolds (total number = 38) and cultured in HARV bioreactors for 28 days. Cell migration, proliferation, osteogenic differentiation, and mineralization were characterized at four selected time points. The same amount of PLAGA scaffolds were used as controls. The incorporation of n-HA did not alter the scaffold degradation pattern. PLAGA/n-HA scaffolds maintained their mechanical integrity throughout the 6 weeks in the dynamic culture environment. HMSCs seeded on PLAGA/n-HA scaffolds showed elevated proliferation, expression of osteogenic phenotypic markers, and mineral deposition as compared with cells seeded on PLAGA scaffolds. HMSCs migrated into the scaffold center with nearly uniform cell and extracellular matrix distribution in the scaffold interior. The combination of PLAGA/n-HA scaffolds with HMSCs in HARV bioreactors may allow for the generation of engineered

Optimization of denitrifying bioreactor performance with agricultural residue-based filter media

USDA-ARS?s Scientific Manuscript database

Denitrification bioreactors are a promising technology for mitigation of nitrate-nitrogen (NO3-N) losses in subsurface drainage water. Bioreactors are constructed with carbon substrates, typically wood chips, to provide a substrate for denitrifying microorganisms. Columns were packed with wood chips...

CASKAD. Manual Mixing in Bioreactor

NASA Image and Video Library

2013-10-05

ISS037-E-005692 (5 Oct. 2013) --- Russian cosmonaut Sergey Ryazanskiy, Expedition 37 flight engineer, prepares to manually mix samples in a Bioreactor for the CASKAD experiment in the Poisk Mini-Research Module 2 (MRM2) of the International Space Station.

CASKAD. Manual Mixing in Bioreactor

NASA Image and Video Library

2013-10-05

ISS037-E-005694 (5 Oct. 2013) --- Russian cosmonaut Sergey Ryazanskiy, Expedition 37 flight engineer, prepares to manually mix samples in a Bioreactor for the CASKAD experiment in the Poisk Mini-Research Module 2 (MRM2) of the International Space Station.

Microgravity

NASA Image and Video Library

1996-01-01

Interior of a Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Microgravity

NASA Image and Video Library

1996-01-01

Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell and with thermal blankets partially removed. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Microgravity

NASA Image and Video Library

1996-01-01

Close-up view of the interior of a NASA Bioreactor shows the plastic plumbing and valves (cylinders at right center) to control fluid flow. The rotating wall vessel is at top center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Microgravity

NASA Image and Video Library

1996-01-01

Biotechnology Refrigerator that preserves samples for use in (or after culturing in) the NASA Bioreactor. The unit is shown extracted from a middeck locker shell. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Measurements of NH 3 and CO 2 with Distributed-Feedback Diode Lasers Near 2.0 m in Bioreactor Vent Gases

NASA Astrophysics Data System (ADS)

Webber, Michael E.; Claps, Ricardo; Englich, Florian V.; Tittel, Frank K.; Jeffries, Jay B.; Hanson, Ronald K.

2001-08-01

Measurements of NH3 and CO2 were made in bioreactor vent gases with distributed-feedback diode-laser sensors operating near 2 m. Calculated spectra of NH3 and CO2 were used to determine the optimum transitions for interrogating with an absorption sensor. For ammonia, a strong and isolated absorption transition at 5016.977 cm-1 was selected for trace gas monitoring. For CO2 , an isolated transition at 5007.787 cm-1 was selected to measure widely varying concentrations [500 parts per million (ppm) to 10% ,] with sufficient signal for low mole fractions and without being optically thick for high mole fractions. Using direct absorption and a 36-m total path-length multipass flow-through cell, we achieved a minimum detectivity of 0.25 ppm for NH3 and 40 ppm for CO2 . We report on the quasi-continuous field measurements of NH3 and CO2 concentration in bioreactor vent gases that were recorded at NASA Johnson Space Center with a portable and automated sensor system over a 45-h data collection window.

Miniature Bioreactor System for Long-Term Cell Culture

NASA Technical Reports Server (NTRS)

Gonda, Steve R.; Kleis, Stanley J.; Geffert, Sandara K.

2010-01-01

A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.

Bioreactors for guiding muscle tissue growth and development.

PubMed

Dennis, R G; Smith, B; Philp, A; Donnelly, K; Baar, K

2009-01-01

Muscle tissue bioreactors are devices which are employed to guide and monitor the development of engineered muscle tissue. These devices have a modern history that can be traced back more than a century, because the key elements of muscle tissue bioreactors have been studied for a very long time. These include barrier isolation and culture of cells, tissues and organs after isolation from a host organism; the provision of various stimuli intended to promote growth and maintain the muscle, such as electrical and mechanical stimulation; and the provision of a perfusate such as culture media or blood derived substances. An accurate appraisal of our current progress in the development of muscle bioreactors can only be made in the context of the history of this endeavor. Modern efforts tend to focus more upon the use of computer control and the application of mechanical strain as a stimulus, as well as substrate surface modifications to induce cellular organization at the early stages of culture of isolated muscle cells.

Microgravity

NASA Image and Video Library

1998-01-01

Cells from kidneys lose some of their special features in conventional culture but form spheres replete with specialized cell microvilli (hair) and synthesize hormones that may be clinically useful. Ground-based research studies have demonstrated that both normal and neoplastic cells and tissues recreate many of the characteristics in the NASA bioreactor that they display in vivo. Proximal kidney tubule cells that normally have rich apically oriented microvilli with intercellular clefts in the kidney do not form any of these structures in conventional two-dimensional monolayer culture. However, when normal proximal renal tubule cells are cultured in three-dimensions in the bioreactor, both the microvilli and the intercellular clefts form. This is important because, when the morphology is recreated, the function is more likely also to be rejuvenated. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC).

Arterial specification of endothelial cells derived from human induced pluripotent stem cells in a biomimetic flow bioreactor.

PubMed

Sivarapatna, Amogh; Ghaedi, Mahboobe; Le, Andrew V; Mendez, Julio J; Qyang, Yibing; Niklason, Laura E

2015-01-01

Endothelial cells (ECs) exist in different microenvironments in vivo, including under different levels of shear stress in arteries versus veins. Standard stem cell differentiation protocols to derive ECs and EC-subtypes from human induced pluripotent stem cells (hiPSCs) generally use growth factors or other soluble factors in an effort to specify cell fate. In this study, a biomimetic flow bioreactor was used to subject hiPSC-derived ECs (hiPSC-ECs) to shear stress to determine the impacts on phenotype and upregulation of markers associated with an anti-thrombotic, anti-inflammatory, arterial-like phenotype. The in vitro bioreactor system was able to efficiently mature hiPSC-ECs into arterial-like cells in 24 h, as demonstrated by qRT-PCR for arterial markers EphrinB2, CXCR4, Conexin40 and Notch1, as well protein-level expression of Notch1 intracellular domain (NICD). Furthermore, the exogenous addition of soluble factors was not able to fully recapitulate this phenotype that was imparted by shear stress exposure. The induction of these phenotypic changes was biomechanically mediated in the shear stress bioreactor. This biomimetic flow bioreactor is an effective means for the differentiation of hiPSC-ECs toward an arterial-like phenotype, and is amenable to scale-up for culturing large quantities of cells for tissue engineering applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

PERFORMANCE OF NORTH AMERICAN BIOREACTOR LANDFILLS: II. CHEMICAL AND BIOLOGICAL CHARACTERISTICS

EPA Science Inventory

The objective of this research was to examine the performance of five North American bioreactor landfills. This paper represents the second of a two part series and addresses biological and chemical aspects of bioreactor performance including gas production and management, and l...

Regulation of mesenchymal stem cell 3D microenvironment: From macro to microfluidic bioreactors.

PubMed

Sart, Sébastien; Agathos, Spiros N; Li, Yan; Ma, Teng

2016-01-01

Human mesenchymal stem cells (hMSCs) have emerged as an important cell type in cell therapy and tissue engineering. In these applications, maintaining the therapeutic properties of hMSCs requires tight control of the culture environments and the structural cell organizations. Bioreactor systems are essential tools to achieve these goals in the clinical-scale expansion and tissue engineering applications. This review summarizes how different bioreactors provide cues to regulate the structure and the chemico-mechanical microenvironment of hMSCs with a focus on 3D organization. In addition to conventional bioreactors, recent advances in microfluidic bioreactors as a novel approach to better control the hMSC microenvironment are also discussed. These advancements highlight the key role of bioreactor systems in preserving hMSC's functional properties by providing dynamic and temporal regulation of in vitro cellular microenvironment. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bioreactors as engineering support to treat cardiac muscle and vascular disease.

PubMed

Massai, Diana; Cerino, Giulia; Gallo, Diego; Pennella, Francesco; Deriu, Marco A; Rodriguez, Andres; Montevecchi, Franco M; Bignardi, Cristina; Audenino, Alberto; Morbiducci, Umberto

2013-01-01

Cardiovascular disease is the leading cause of morbidity and mortality in the Western World. The inability of fully differentiated, load-bearing cardiovascular tissues to in vivo regenerate and the limitations of the current treatment therapies greatly motivate the efforts of cardiovascular tissue engineering to become an effective clinical strategy for injured heart and vessels. For the effective production of organized and functional cardiovascular engineered constructs in vitro, a suitable dynamic environment is essential, and can be achieved and maintained within bioreactors. Bioreactors are technological devices that, while monitoring and controlling the culture environment and stimulating the construct, attempt to mimic the physiological milieu. In this study, a review of the current state of the art of bioreactor solutions for cardiovascular tissue engineering is presented, with emphasis on bioreactors and biophysical stimuli adopted for investigating the mechanisms influencing cardiovascular tissue development, and for eventually generating suitable cardiovascular tissue replacements.

Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor

PubMed Central

2012-01-01

Background To simplify clinical scale lymphocyte expansions, we investigated the use of the WAVE®, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. Methods We have developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant numbers of cells for our adoptive cell transfer clinical protocols. Results TIL and genetically modified PBL were rapidly expanded to clinically relevant scales in both static bags and the WAVE bioreactor. Both bioreactors produced comparable numbers of cells; however the cultures generated in the WAVE bioreactor had a higher percentage of CD4+ cells and had a less activated phenotype. Conclusions The WAVE bioreactor simplifies the process of rapidly expanding tumor reactive lymphocytes under GMP conditions, and provides an alternate approach to cell generation for ACT protocols. PMID:22475724

NASA's Autonomous Formation Flying Technology Demonstration, Earth Observing-1(EO-1)

NASA Technical Reports Server (NTRS)

Folta, David; Bristow, John; Hawkins, Albin; Dell, Greg

2002-01-01

NASA's first autonomous formation flying mission, the New Millennium Program's (NMP) Earth Observing-1 (EO-1) spacecraft, recently completed its principal goal of demonstrating advanced formation control technology. This paper provides an overview of the evolution of an onboard system that was developed originally as a ground mission planning and operations tool. We discuss the Goddard Space Flight Center s formation flying algorithm, the onboard flight design and its implementation, the interface and functionality of the onboard system, and the implementation of a Kalman filter based GPS data smoother. A number of safeguards that allow the incremental phasing in of autonomy and alleviate the potential for mission-impacting anomalies from the on- board autonomous system are discussed. A comparison of the maneuvers planned onboard using the EO-1 autonomous control system to those from the operational ground-based maneuver planning system is presented to quantify our success. The maneuvers discussed encompass reactionary and routine formation maintenance. Definitive orbital data is presented that verifies all formation flying requirements.

Review of nonconventional bioreactor technology

DOE Office of Scientific and Technical Information (OSTI.GOV)

Turick, C.E.; Mcllwain, M.E.

1993-09-01

Biotechnology will significantly affect many industrial sectors in the future. Industrial sectors that will be affected include pharmaceutical, chemical, fuel, agricultural, and environmental remediation. Future research is needed to improve bioprocessing efficiency and cost-effectiveness in order to compete with traditional technologies. This report describes recent advances in bioprocess technologies and bioreactor designs and relates them to problems encountered in many industrial bioprocessing operations. The primary focus is directed towards increasing gas and vapor transfer for enhanced bioprocess kinetics as well as unproved by-product separation and removal. The advantages and disadvantages of various conceptual designs such as hollow-fiber, gas-phase, hyperbaric/hypobaric, andmore » electrochemical bioreactors are also discussed. Specific applications that are intended for improved bioprocesses include coal desulfurization, coal liquefaction, soil bioremediation, biomass conversion to marketable chemicals, biomining, and biohydrometallurgy as well as bioprocessing of gases and vapors.« less

Bioreactor production of secondary metabolites from cell cultures of periwinkle and sandalwood.

PubMed

Valluri, Jagan V

2009-01-01

A bench-top bioreactor allowing continuous extraction of secondary metabolites is designed for Catharanthus roseus L. (G.) Don (periwinkle) and Santalum album L. (sandalwood) plant cell suspensions. Periwinkle cell cultures are exposed to biotic elicitors (Aspergillus niger, crude chitin) and abiotic elicitors (mannitol, methyl jasmonate) to induce alkaloid production. Whereas most of the biotic elicitors are effective when added on day 15 of culture, the abiotic elicitors are effective when added on day 20. The use of trans-cinnamic acid, an inhibitor of phenylalanine ammonia lyase (PAL) activity, results in significant increase in the alkaloid production of periwinkle cell cultures. Exposure of the cells to mannitol-induced osmotic stress produced marked increment in the total alkaloid production. When biotic and abiotic stress treatments are applied sequentially, an additive effect in alkaloid accumulation is observed. Although no essential oils are detected, secondary metabolites in the form of phenolics are produced by the sandalwood cell cultures in the bioreactor environment. The use of morphologic modification such as organ cultures and transformed cultures is believed to be required for both production and storage of essential oil constituents in sandalwood. The present chapter demonstrates that periwinkle and sandalwood cell suspensions could be developed and successfully cultured in a modified air-lift bioreactor. The exploitation of variant cell strains and biotransformation of added precursors can certainly improve the use of periwinkle and sandalwood cell cultures for the bioproduction of desired compounds.

Membrane bioreactor technology: A novel approach to the treatment of compost leachate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, Kayleigh; Ghoshdastidar, Avik J.; Hanmore, Jillian

Highlights: • First membrane bioreactor treatment method for compost leachate. • No chemical additive or UV radiation source in this new biological method. • Removal rates of more than 99% for organics and ammonium were achieved. • Heavy metals were reduced by at least 82.7% except copper. - Abstract: Compost leachate forms during the composting process of organic material. It is rich in oxidizable organics, ammonia and metals, which pose a risk to the environment if released without proper treatment. An innovative method based on the membrane bioreactor (MBR) technology was developed to treat compost leachate over 39 days. Watermore » quality parameters, such as pH, dissolved oxygen, ammonia, nitrate, nitrite and chemical oxygen demand (COD) were measured daily. Concentrations of caffeine and metals were measured over the course of the experiment using gas chromatography – mass spectrometry (GC/MS) and inductively coupled plasma – mass spectrometry (ICP–MS) respectively. A decrease of more than 99% was achieved for a COD of 116 g/L in the initial leachate. Ammonia was decreased from 2720 mg/L to 0.046 mg/L, while the nitrate concentration in the effluent rose to 710 mg/L. The bacteria in the MBR system adjusted to the presence of the leachate, and increased 4 orders of magnitude. Heavy metals were removed by at least 82.7% except copper. These successful results demonstrated the membrane bioreactor technology is feasible, efficient method for the treatment of compost leachate.« less

Biological treatment of mixtures of toluene and n-hexane vapours in a hollow fibre membrane bioreactor.

PubMed

Zhao, Kang; Xiu, Guangli; Xu, Lihang; Zhang, Danian; Zhang, Xiaofeng; Deshusses, Marc A

2011-04-01

Membrane bioreactors are gaining interest for the control of contaminated air streams. In this study, the removal of toluene and n-hexane vapours in a hollow fibre membrane bioreactor (HFMB) was investigated. The focus was on quantifying the possible interactions occurring during the simultaneous biotreatment of the two volatile pollutants. Two lab-scale units fitted with microporous polypropylene hollow fibre membranes were connected in series and inoculated with activated sludge. Contaminated air was passed through the lumen at gas residence times ranging from 2.3 to 9.4 s while a pollutant-degrading biofilm developed on the shell side of the fibres. When toluene was treated alone, very high elimination capacities (up to 750 g m(-3) h(-1) based on lumen volume, or 1.25 g m(-2) h(-1) when normalized by the hollow fibre membrane area) were reached. When toluene and hexane were treated simultaneously, toluene biodegradation was partially inhibited by n-hexane, resulting in lower toluene removal rates. On the other hand, hexane removal was only marginally affected by the presence of toluene and was degraded at very high rates (upwards of 440 g m(-3) h(-1) or 0.73 g m(-2) h(-1) without breakthrough). Overall, this study demonstrates that mixtures of toluene and n-hexane vapours can be effectively removed in hollow fibre membrane bioreactors and that complex biological interactions may affect one or more of the pollutants undergoing treatment in gas-phase membrane bioreactors.

Electrical stimulation for enhanced denitrification in woodchip bioreactors: Opportunities and challenges

USDA-ARS?s Scientific Manuscript database

Woodchip bioreactors are being implemented for the removal of nitrates in groundwater and tile water drainage. However, low nitrate removals in denitrifying woodchip bioreactors have been observed for short hydraulic retention time (HRT) and low water temperature (< 10ºC). One potential approach to ...

Bioreactor design for successive culture of anchorage-dependent cells operated in an automated manner.

PubMed

Kino-Oka, Masahiro; Ogawa, Natsuki; Umegaki, Ryota; Taya, Masahito

2005-01-01

A novel bioreactor system was designed to perform a series of batchwise cultures of anchorage-dependent cells by means of automated operations of medium change and passage for cell transfer. The experimental data on contamination frequency ensured the biological cleanliness in the bioreactor system, which facilitated the operations in a closed environment, as compared with that in flask culture system with manual handlings. In addition, the tools for growth prediction (based on growth kinetics) and real-time growth monitoring by measurement of medium components (based on small-volume analyzing machinery) were installed into the bioreactor system to schedule the operations of medium change and passage and to confirm that culture proceeds as scheduled, respectively. The successive culture of anchorage-dependent cells was conducted with the bioreactor running in an automated way. The automated bioreactor gave a successful culture performance with fair accordance to preset scheduling based on the information in the latest subculture, realizing 79- fold cell expansion for 169 h. In addition, the correlation factor between experimental data and scheduled values through the bioreactor performance was 0.998. It was concluded that the proposed bioreactor with the integration of the prediction and monitoring tools could offer a feasible system for the manufacturing process of cultured tissue products.

In situ Raman spectroscopy for simultaneous monitoring of multiple process parameters in mammalian cell culture bioreactors.

PubMed

Whelan, Jessica; Craven, Stephen; Glennon, Brian

2012-01-01

In this study, the application of Raman spectroscopy to the simultaneous quantitative determination of glucose, glutamine, lactate, ammonia, glutamate, total cell density (TCD), and viable cell density (VCD) in a CHO fed-batch process was demonstrated in situ in 3 L and 15 L bioreactors. Spectral preprocessing and partial least squares (PLS) regression were used to correlate spectral data with off-line reference data. Separate PLS calibration models were developed for each analyte at the 3 L laboratory bioreactor scale before assessing its transferability to the same bioprocess conducted at the 15 L pilot scale. PLS calibration models were successfully developed for all analytes bar VCD and transferred to the 15 L scale. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

CFD of mixing of multi-phase flow in a bioreactor using population balance model.

PubMed

Sarkar, Jayati; Shekhawat, Lalita Kanwar; Loomba, Varun; Rathore, Anurag S

2016-05-01

Mixing in bioreactors is known to be crucial for achieving efficient mass and heat transfer, both of which thereby impact not only growth of cells but also product quality. In a typical bioreactor, the rate of transport of oxygen from air is the limiting factor. While higher impeller speeds can enhance mixing, they can also cause severe cell damage. Hence, it is crucial to understand the hydrodynamics in a bioreactor to achieve optimal performance. This article presents a novel approach involving use of computational fluid dynamics (CFD) to model the hydrodynamics of an aerated stirred bioreactor for production of a monoclonal antibody therapeutic via mammalian cell culture. This is achieved by estimating the volume averaged mass transfer coefficient (kL a) under varying conditions of the process parameters. The process parameters that have been examined include the impeller rotational speed and the flow rate of the incoming gas through the sparger inlet. To undermine the two-phase flow and turbulence, an Eulerian-Eulerian multiphase model and k-ε turbulence model have been used, respectively. These have further been coupled with population balance model to incorporate the various interphase interactions that lead to coalescence and breakage of bubbles. We have successfully demonstrated the utility of CFD as a tool to predict size distribution of bubbles as a function of process parameters and an efficient approach for obtaining optimized mixing conditions in the reactor. The proposed approach is significantly time and resource efficient when compared to the hit and trial, all experimental approach that is presently used. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:613-628, 2016. © 2016 American Institute of Chemical Engineers.

Enhanced Denitrification in Roadside Ditches with Bioreactors

NASA Astrophysics Data System (ADS)

Pluer, W.; Schneider, R.; Walter, M. T.

2016-12-01

Nitrate (NO3) pollution remains a water quality problem in agriculture-dominated watersheds despite decades of research and concerted efforts. Excess NO3 causes eutrophication in estuarine and marine ecosystems far downstream of the pollution source. Denitrification reduces NO3 to inert dinitrogen gas; this process occurs naturally in saturated areas of the landscape but this rate cannot keep up with the runoff rate due to fertilizer and manure applications. Researchers developed denitrifying bioreactors as a solution to encourage denitrification at the field level. Denitrifying bioreactors remove NO3 at a significantly higher rate (>2 g N m-2 d-1) than natural systems such as wetlands (<0.5 g N m-2 d-1). Most current designs of denitrifying bioreactors necessitate connection with tile drainage as the inflow source of water and NO3. It also requires a portion of farmland (typically <1% of field area is needed) which farmers can be reluctant to relinquish. Meanwhile, road ditches commonly run along agricultural fields, channeling runoff and NO3 to surface water. Because the ditches are designed to avoid flooding, they channel water rapidly and minimize time and contact with soil microbes for denitrification (denitrification rates in ditches are typically <0.1 g N m-2 d-1). Modified denitrifying bioreactors placed in road ditches could provide high NO3 removal in already marginal land, especially at baseflow conditions. A pilot study of this shows instantaneous NO3 removal rates up to 110 g N m-2 d-1 in the first year. Continued results similar to this pilot study and wider application could significantly increase ditch denitrification and help mitigate NO3 pollution.

Quantitative analysis of microbial biomass yield in aerobic bioreactor.

PubMed

Watanabe, Osamu; Isoda, Satoru

2013-12-01

We have studied the integrated model of reaction rate equations with thermal energy balance in aerobic bioreactor for food waste decomposition and showed that the integrated model has the capability both of monitoring microbial activity in real time and of analyzing biodegradation kinetics and thermal-hydrodynamic properties. On the other hand, concerning microbial metabolism, it was known that balancing catabolic reactions with anabolic reactions in terms of energy and electron flow provides stoichiometric metabolic reactions and enables the estimation of microbial biomass yield (stoichiometric reaction model). We have studied a method for estimating real-time microbial biomass yield in the bioreactor during food waste decomposition by combining the integrated model with the stoichiometric reaction model. As a result, it was found that the time course of microbial biomass yield in the bioreactor during decomposition can be evaluated using the operational data of the bioreactor (weight of input food waste and bed temperature) by the combined model. The combined model can be applied to manage a food waste decomposition not only for controlling system operation to keep microbial activity stable, but also for producing value-added products such as compost on optimum condition. Copyright © 2013 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Use of ATP to characterize biomass viability in freely suspended and immobilized cell bioreactors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gikas, P.; Livingston, A.G.

1993-12-01

This work describes investigations into the viability of cells growing on 3,4-dichloroaniline (34DCA). Two bio-reactors are employed for microbial growth, a continuous stirred tank (CST) bioreactor with a 2-L working volume, and a three-phase air lift (TPAL) bioreactor with a 3-L working volume. Experiments have been performed at several dilution rates between 0.027 and 0.115 h[sup [minus]1] in the CST bioreactor and between 0.111 and 0.500h[sup [minus]1] in the TPAL bioreactor. The specific ATP concentration was calculated at each dilution rate in the suspended biomass in both bioreactors as well as in the immobilized biomass in the TPAL bioreactor. Themore » cultures were inspected under an electron microscope to monitor compositional changes. Results from the CST bioreactor showed that the biomass-specific ATP concentration increases from 0.44 to 1.86 mg ATP g[sup [minus]1] dry weight (dw) as dilution rate increases from 0.027 to 0.115 h[sup [minus]1]. At this upper dilution rate the cells were washed out. The specific ATP concentration reached a limiting average value of 1.73 mg ATP g[sup [minus]1] dw, which is assumed to be the quantity of ATP in 100% viable biomass, In the TPAL bioreactor, the ATP level increased with dilution rat in both the immobilized and suspended biomass. The specific ATP concentration in the immobilized biomass increased from approximately 0.051 mg ATP g[sup [minus]1] dw at dilution rates between 0.111 and 0.200 h[sup [minus]1] to approximately 0.119 mg ATP g[sup [minus]1] dw at dilution rates between 0.300 and 0.500 h[sup [minus]1].« less

Replaceable Sensor System for Bioreactor Monitoring

NASA Technical Reports Server (NTRS)

Mayo, Mike; Savoy, Steve; Bruno, John

2006-01-01

A sensor system was proposed that would monitor spaceflight bioreactor parameters. Not only will this technology be invaluable in the space program for which it was developed, it will find applications in medical science and industrial laboratories as well. Using frequency-domain-based fluorescence lifetime technology, the sensor system will be able to detect changes in fluorescence lifetime quenching that results from displacement of fluorophorelabeled receptors bound to target ligands. This device will be used to monitor and regulate bioreactor parameters including glucose, pH, oxygen pressure (pO2), and carbon dioxide pressure (pCO2). Moreover, these biosensor fluorophore receptor-quenching complexes can be designed to further detect and monitor for potential biohazards, bioproducts, or bioimpurities. Biosensors used to detect biological fluid constituents have already been developed that employ a number of strategies, including invasive microelectrodes (e.g., dark electrodes), optical techniques including fluorescence, and membrane permeable systems based on osmotic pressure. Yet the longevity of any of these sensors does not meet the demands of extended use in spacecraft habitat or bioreactor monitoring. It was therefore necessary to develop a sensor platform that could determine not only fluid variables such as glucose concentration, pO2, pCO2, and pH but can also regulate these fluid variables with controlled feedback loop.

Use of bioreactors for culturing human retinal organoids improves photoreceptor yields.

PubMed

Ovando-Roche, Patrick; West, Emma L; Branch, Matthew J; Sampson, Robert D; Fernando, Milan; Munro, Peter; Georgiadis, Anastasios; Rizzi, Matteo; Kloc, Magdalena; Naeem, Arifa; Ribeiro, Joana; Smith, Alexander J; Gonzalez-Cordero, Anai; Ali, Robin R

2018-06-13

The use of human pluripotent stem cell-derived retinal cells for cell therapy strategies and disease modelling relies on the ability to obtain healthy and organised retinal tissue in sufficient quantities. Generating such tissue is a lengthy process, often taking over 6 months of cell culture, and current approaches do not always generate large quantities of the major retinal cell types required. We adapted our previously described differentiation protocol to investigate the use of stirred-tank bioreactors. We used immunohistochemistry, flow cytometry and electron microscopy to characterise retinal organoids grown in standard and bioreactor culture conditions. Our analysis revealed that the use of bioreactors results in improved laminar stratification as well as an increase in the yield of photoreceptor cells bearing cilia and nascent outer-segment-like structures. Bioreactors represent a promising platform for scaling up the manufacture of retinal cells for use in disease modelling, drug screening and cell transplantation studies.

NASA Social

NASA Image and Video Library

2011-05-18

Gwynne Shotwell, President of SpaceX, speaks during a NASA Social, Friday, May 18, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

NASA Social

NASA Image and Video Library

2012-05-18

Models of various rockets line a table at a NASA Social, Friday, May 18, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

Recent Advances in Cartilage Tissue Engineering: From the Choice of Cell Sources to the Use of Bioreactors

NASA Astrophysics Data System (ADS)

Martin, Ivan; Démarteau, Olivier; Braccini, Alessandra

Grafting engineered cartilage tissues represents a promising approach for the repair of joint injuries. Recent animal experiments have demonstrated that tissues engineered by culturing chondrocytes on 3D scaffolds in bioreactors provide functional templates for orderly repair of large osteochondral lesions. To date, however, a reproducible generation of uniform cartilage tissues of predefined size starting from adult human cells has not been achieved. In this paper we review some of the recent advances and challenges ahead in the identification of appropriate (i) cell sources, (ii) bioactive factors, (iii) 3D scaffolds and (iv) bioreactors for human cartilage tissue engineering. We also present an example of how integrated efforts in these different areas can help addressing fundamental questions and advancing the field of cartilage tissue engineering towards clinical use. The presented experiment demonstrates that human nasal chondrocytes are responsive to dynamic loading and thus could be further investigated as a cell source for implantation in a joint environment.

Studies of Cell-Mediated Immunity Against Immune Disorders Using Synthetic Peptides and Rotating Bioreactor System

NASA Technical Reports Server (NTRS)

Sastry, Jagannadha K.

1997-01-01

Our proposed experiments included: (1) immunzing mice with synthetic peptides; (2) preparing spleen and lymph node cells; (3) growing them under conventional conditions as well as in the rotatory vessel in appropriate medium reconstituting with synthetic peptides and/or cytokines as needed; and (4) comparing at regular time intervals the specific CTL activity as well as helper T-cell activity (in terms of both proliferative responses and cytokine production) using established procedures in my laboratory. We further proposed that once we demonstrated the merit of rotatory vessel technology to achieve desired results, these studies would be expanded to include immune cells from non-human primates (rhesus monkeys and chimpanzees) and also humans. We conducted a number of experiments to determine CTL induction by the synthetic peptides corresponding to antigenic proteins in HIV and HPV in different mouse strains that express MHC haplotypes H-2b or H-2d. We immunized mice with 100 ug of the synthetic peptide, suspended in sterile water, and emulsified in CFA (1:1). The immune lymph node cells obtained after 7 days were restimulated by culturing in T25 flask, HARV-10, or STLV-50, in the presence of the peptide at 20 ug/ml. The results from the 5'Cr-release assay consistently revealed complete abrogation of CTL activity of cells grown in the bioreactors (both HARV and STLV), while significant antigen-specific CTL activity was observed with cells cultured in tissue culture flasks. Thus, overall the data we generated in this study proved the usefulness of the NASA-developed developed technology for understanding the known immune deficiency during space travel. Additionally, this ex vivo microgravity technology since it mimics effectively the in vivo situation, it is also useful in understanding immune disorders in general. Thus, our proposed studies in TMC-NASA contract round II application benefit from data generated in this TMC-NASA contract round I study.

Reduced Differentiation Efficiency of Murine Embryonic Stem Cells in Stirred Suspension Bioreactors

PubMed Central

Taiani, Jaymi T.; Krawetz, Roman J.; zur Nieden, Nicole I.; Wu, Yiru Elizabeth; Kallos, Michael S.; Matyas, John R.

2010-01-01

The use of embryonic stem cells (ESCs) for regenerative medicine has generated increased attention due to the favorable attributes of these cells; namely, they are pluripotent and possess long-term self-renewal capacity. The initial aims of the present study were: (i) to use stirred suspension bioreactors to expand and differentiate ESCs into osteogenic and chondrogenic cell types and (ii) to explore if these ESC-derived cells influenced skeletal healing in an in vivo fracture model. We show that differentiation protocols used in static culture are insufficient when applied directly to suspension culture bioreactors. Moreover, when bioreactor-differentiated cells are transplanted into a burr-hole defect in bone, severe disruption of the bone architecture was noted at the fracture site, as determined by microcomputed tomography (microCT) imaging and histopathology. Further characterization of the bioreactor-differentiated cultures revealed that a subpopulation of cells in the resulting aggregates expressed the pluripotency marker Oct-4 in the nucleus. Nuclear Oct-4 expression persisted even after 30 days of culture in the absence of leukemia inhibitory factor (LIF). Remarkably, and unlike ESCs differentiated into skeletal cell types in static cultures, bioreactor-differentiated aggregates implanted subcutaneously into SCID mice formed teratomas. The development of effective ESC differentiation protocols for suspension bioreactors will require a more complete understanding of the environmental conditions within these culture systems and the influence that these conditions have on the regulation of pluripotency and differentiation in ESCs. PMID:19775198

NASA in the Park, 2018

NASA Image and Video Library

2018-06-20

NASA in the Park on June 16 in Huntsville featured more than 60 exhibits and demonstrations by NASA experts, as well as performances by Marshall musicians, educational opportunities, games and hands-on activities for all ages. National Space Club members help demonstrate propulsion principles at NASA in the Park.

Thinking beyond the Bioreactor Box: Incorporating Stream Ecology into Edge-of-Field Nitrate Management.

PubMed

Goeller, Brandon C; Febria, Catherine M; Harding, Jon S; McIntosh, Angus R

2016-05-01

Around the world, artificially drained agricultural lands are significant sources of reactive nitrogen to stream ecosystems, creating substantial stream health problems. One management strategy is the deployment of denitrification enhancement tools. Here, we evaluate the factors affecting the potential of denitrifying bioreactors to improve stream health and ecosystem services. The performance of bioreactors and the structure and functioning of stream biotic communities are linked by environmental parameters like dissolved oxygen and nitrate-nitrogen concentrations, dissolved organic carbon availability, flow and temperature regimes, and fine sediment accumulations. However, evidence of bioreactors' ability to improve waterway health and ecosystem services is lacking. To improve the potential of bioreactors to enhance desirable stream ecosystem functioning, future assessments of field-scale bioreactors should evaluate the influences of bioreactor performance on ecological indicators such as primary production, organic matter processing, stream metabolism, and invertebrate and fish assemblage structure and function. These stream health impact assessments should be conducted at ecologically relevant spatial and temporal scales. Bioreactors have great potential to make significant contributions to improving water quality, stream health, and ecosystem services if they are tailored to site-specific conditions and implemented strategically with land-based and stream-based mitigation tools within watersheds. This will involve combining economic, logistical, and ecological information in their implementation. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Microgravity

NASA Image and Video Library

1996-01-01

Exterior view of the NASA Bioreactor Engineering Development Unit flown on Mir. The rotating wall vessel is behind the window on the face of the large module. Control electronics are in the module at left; gas supply and cooling fans are in the module at back. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Tissue engineering bioreactor systems for applying physical and electrical stimulations to cells.

PubMed

Jin, GyuHyun; Yang, Gi-Hoon; Kim, GeunHyung

2015-05-01

Bioreactor systems in tissue engineering applications provide various types of stimulation to mimic the tissues in vitro and in vivo. Various bioreactors have been designed to induce high cellular activities, including initial cell attachment, cell growth, and differentiation. Although cell-stimulation processes exert mostly positive effects on cellular responses, in some cases such stimulation can also have a negative effect on cultured cells. In this review, we discuss various types of bioreactor and the positive and negative effects of stimulation (physical, chemical, and electrical) on various cultured cell types. © 2014 Wiley Periodicals, Inc.

The Flow in a Model Rotating-Wall Bioreactor.

NASA Astrophysics Data System (ADS)

Smith, Marc K.; Neitzel, G. Paul

1997-11-01

Aggregates of mammalian cells can be grown on artificial polymer constructs in a reactor vessel in order to produce high-quality tissue for medical applications. The growth and differentiation of these cells is greatly affected by the fluid flow and mass transfer within the bioreactor. The surface shear stress on the constructs is an especially important quantity of interest. Here, we consider a bioreactor in the form of two concentric, independently-rotating cylinders with the axis of rotation in a horizontal plane. We shall examine the flow around a model tissue construct in the form of a disk fixed in the flow produced by the rotating walls of the bioreactor. Using CFD techniques, we shall determine the flow field and the surface shear stress distribution on the construct as a function of the wall velocities, the Reynolds number of the flow, and the construct size and position. The results will be compared to the PIV measurements of this system reported by Brown & Neitzel(1997 Meeting of the APS/DFD.).

A novel axial-stress bioreactor system combined with a substance exchanger for tissue engineering of 3D constructs.

PubMed

Li, Song-Tao; Liu, Yong; Zhou, Qiang; Lue, Ren-Fa; Song, Lei; Dong, Shi-Wu; Guo, Ping; Kopjar, Branko

2014-03-01

This study introduced a prototype of an axial-stress bioreactor system that supports long-term growth and development of engineered tissues. The main features of this bioreactor are an integrated substance exchanger and feedback control of pH and PO₂. A 21-day study was conducted to validate the system's ability to maintain a stable environment, while remaining sterile. Our results showed that the pH, PO₂, and nutrient (glucose) remained balanced at appropriate levels, while metabolic waste (lactic acid) was removed. No bacteria or fungi were detected in the system or tissue; thus, demonstrating that it was sterile. These data indicate the bioreactor's strong potential for long-term tissue culture. To explore this idea, the effect of dynamic culture, including cyclic compression and automatic substance exchange, on mouse bone-marrow mesenchymal stem cells (BMSCs) seeded in decalcified bone matrix was studied using the bioreactor prototype. Histological sections of the engineered tissues showed higher cell densities in scaffolds in dynamic culture compared to those in static culture, while cell cycle analysis showed that dynamic culture promoted BMSC proliferation (proliferation index, PI=34.02±1.77) more effectively than static culture (PI=26.66±1.81). The results from a methyl thiazolyl tetrazolium assay were consistent with the loading experimental data. Furthermore, elevated alkaline phosphatase activity and calcium content were observed in dynamic condition compared to static culture. In conclusion, this bioreactor system supplies a method of modulating the pH and PO₂ in defined ranges with only small fluctuations; it can be used as a physiological or pathological analog. Automatic control of the environment is a practical solution for long-term, steady-state culture for future commercialization.

Kinetics of heavy metal removal in a suspended and immobilized bioreactors

NASA Astrophysics Data System (ADS)

Kutty, S. R. M.; Ezechi, E. H.; Khaw, S. G.; Lai, C. L.; Isa, M. H.

2017-06-01

The capacity of microorganisms to remove heavy metal from wastewater has been a subject of diverse interest. Whereas some heavy metals are essential for effective microbial activity, some heavy metals could be toxic to the microorganisms at concentrations higher than their minimal inhibitory limit. The kinetics of Zn2+ removal from aqueous solution was evaluated in terms of substrate removal rate for two identical suspended and immobilized bioreactors. The suspended growth bioreactor was used as a control system (CS) and contains only biomass. The immobilized bioreactor (IB) contains both biomass and microwave incinerated rice husk ash (MIRHA). The bioreactors were operated at a fixed HRT of 29.1 hours, whereas Zn2+ influent concentration was varied in the range of 0.5, 1, 2, 5, 10 and 15 mg/L. At steady state conditions, the results show that Zn2+ removal was in the range of 72, 75, 72.5, 68.2, 70.3 and 58.7% for CS, whereas it was in the range of 88, 90, 83, 88.6, 86.2 and 83.7% for IB. The substrate removal rate was found as 1.1856 g/L.d for CS and 4.2693 g/L.d for IB. The results clearly show that Zn2+ removal was more favorable in IB, indicating that the performance of the bioreactor was enhanced by the addition of MIRHA.

Oxygen mass transfer in a stirred tank bioreactor using different impeller configurations for environmental purposes

PubMed Central

2013-01-01

In this study, a miniature stirred tank bioreactor was designed for treatment of waste gas containing benzene, toluene and xylene. Oxygen mass transfer characteristics for various twin and single-impeller systems were investigated for 6 configurations in a vessel with 10 cm of inner diameter and working volume of 1.77L. Three types of impellers, namely, Rushton turbine, Pitched 4blades and Pitched 2blades impellers with downward pumping have been used. Deionized water was used as a liquid phase. With respect to other independent variables such as agitation speed, aeration rate, type of sparger, number of impellers, the relative performance of these impellers was assessed by comparing the values of (KLa) as a key parameter. Based on the experimental data, empirical correlations as a function of the operational conditions have been proposed, to study the oxygen transfer rates from air bubbles generated in the bioreactor. It was shown that twin Rushton turbine configuration demonstrates superior performance (23% to 77% enhancement in KLa) compared with other impeller compositions and that sparger type has negligible effect on oxygen mass transfer rate. Agitation speeds of 400 to 800 rpm were the most efficient speeds for oxygen mass transfer in the stirred bioreactor. PMID:23369581

NASA Social

NASA Image and Video Library

2012-05-18

Participants with the NASA Social stand together, Friday, May 18, 2012, in front of the Vehicle Assembly Building (VAB) at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

Application of Pi Preform Composite Joints in Fabrication of NASA Composite Crew Module Demonstration Structure

NASA Technical Reports Server (NTRS)

Higgins, John E.; Pelham, Larry

2008-01-01

This paper will describe unique and extensive use of pre-woven and impregnated pi cross-sections in fabrication of a carbon composite demonstration structure for the Composite Crew Module (CCM) Program. The program is managed by the NASA Safety and Engineering Center with participants from ten NASA Centers and AFRL. Multiple aerospace contractors are participating in the design development, tooling and fabrication effort as well. The goal of the program is to develop an agency wide design team for composite habitable spacecraft. The specific goals for this development project are: a) To gain hands on experience in design, building and testing a composite crew module. b) To validate key assumptions by resolving composite spacecraft design details through fabrication and testing of hardware. This paper will focus on the design and fabrication issues supporting selection of the Lockheed Martin patented Pi pre-form to provide sound composite joints a numerous locations in the structure. This abstract is based on Preliminary Design data. The final design will continue to evolve through the fall of 2007 with fabrication mostly completed by conference date.

Lidocaine/monoethylglycinexylidide test, galactose elimination test, and sorbitol elimination test for metabolic assessment of liver cell bioreactors.

PubMed

Gerlach, Jörg C; Brayfield, Candace; Puhl, Gero; Borneman, Reiner; Müller, Christian; Schmelzer, Eva; Zeilinger, Katrin

2010-06-01

Various metabolic tests were compared for the performance characterization of a liver cell bioreactor as a routine function assessment of cultures in a standby for patient application in clinical studies. Everyday quality assessment (QA) is essential to ensure a continuous level of cellular functional capacity in the development of hepatic progenitor cell expansion systems providing cells for regenerative medicine research; it is also of interest to meet safety requirements in bioartificial extracorporeal liver support systems under clinical evaluation. Quality criteria for the description of bioreactor cultures were developed using primary porcine liver cells as a model. Porcine liver cells isolated by collagenase perfusion with an average of 3 x 10(9) primary cells were used in 39 bioreactors for culture periods up to 33 days. Measurements of monoethylglycinexylidide synthesis and elimination of lidocaine, galactose elimination, and sorbitol elimination proved to be useful for routine QA of primary liver cell cultures. We demonstrate two methods for dispensing test substances, bolus administration and continuous, steady-state administration. Bolus test data were grouped in Standard, Therapy, Infection/Contamination, and Cell-free control groups. Statistical analyses show significant differences among all groups for every test substance. Post hoc comparisons indicated significant differences between Standard and Cell-free groups for all elimination parameters. For continuous tests, results were categorized according to number of culture days and time-dependent changes were analyzed. Continuous administration enables a better view of culture health and the time dependency of cellular function, whereas bolus administration is more flexible. Both procedures can be used to define cell function. Assessment of cellular function and bioreactor quality can contribute significantly to the quality of experimental or clinical studies in the field of hepatic bioreactor

A comparative study on the anaerobic membrane bioreactor performance during the treatment of domestic wastewaters of various origins.

PubMed

Saddoud, A; Ellouze, M; Dhouib, A; Sayadi, S

2006-09-01

This study examined the practical performance of a cross-flow ultrafiltration membrane coupled to an anaerobic bioreactor, for treatment of raw domestic wastewater (RDW), at a pilot-scale plant. Wastewaters used in this study originated from two different domestic wastewater treatment plans (DWTPs) (Sfax and Ksour Essef). During the treatment in the membrane bioreactor (MBR) of the RDW originating from Sfax DWTP, the bioreactor did not reach its stationary phase because the anaerobic biomass was unable to adapt to the wastewater. This was explained by the considerable fluctuations in the domestic wastewater composition and a possible contamination of Sfax wastewater by industrial discharges. However, the treatment of RDW originating from Ksour Essef (DWTP) was successful. In both cases, the treatment led to a total removal of all tested pathogens. The quality of treated wastewater fits largely with WHO guidelines for unrestricted irrigation. The phytotoxicity and the microtoxicity tests, using Lepidium sativum and Vibrio fischeri respectively, demonstrated that wastewater from Sfax exhibited higher toxicity than that from Ksour Sssef.

Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

NASA Technical Reports Server (NTRS)

Kleis, Stanley J. (Inventor); Geffert, Sandra K. (Inventor); Gonda, Steve R. (Inventor)

2015-01-01

A bioreactor and method that permits continuous and simultaneous short, moderate, or long term cell culturing of one or more cell types or tissue in a laminar flow configuration is disclosed, where the bioreactor supports at least two laminar flow zones, which are isolated by laminar flow without the need for physical barriers between the zones. The bioreactors of this invention are ideally suited for studying short, moderate and long term studies of cell cultures and the response of cell cultures to one or more stressors such as pharmaceuticals, hypoxia, pathogens, or any other stressor. The bioreactors of this invention are also ideally suited for short, moderate or long term cell culturing with periodic cell harvesting and/or medium processing for secreted cellular components.

Sensing in tissue bioreactors

NASA Astrophysics Data System (ADS)

Rolfe, P.

2006-03-01

Specialized sensing and measurement instruments are under development to aid the controlled culture of cells in bioreactors for the fabrication of biological tissues. Precisely defined physical and chemical conditions are needed for the correct culture of the many cell-tissue types now being studied, including chondrocytes (cartilage), vascular endothelial cells and smooth muscle cells (blood vessels), fibroblasts, hepatocytes (liver) and receptor neurones. Cell and tissue culture processes are dynamic and therefore, optimal control requires monitoring of the key process variables. Chemical and physical sensing is approached in this paper with the aim of enabling automatic optimal control, based on classical cell growth models, to be achieved. Non-invasive sensing is performed via the bioreactor wall, invasive sensing with probes placed inside the cell culture chamber and indirect monitoring using analysis within a shunt or a sampling chamber. Electroanalytical and photonics-based systems are described. Chemical sensing for gases, ions, metabolites, certain hormones and proteins, is under development. Spectroscopic analysis of the culture medium is used for measurement of glucose and for proteins that are markers of cell biosynthetic behaviour. Optical interrogation of cells and tissues is also investigated for structural analysis based on scatter.

Generation of Neural Progenitor Spheres from Human Pluripotent Stem Cells in a Suspension Bioreactor.

PubMed

Yan, Yuanwei; Song, Liqing; Tsai, Ang-Chen; Ma, Teng; Li, Yan

2016-01-01

Conventional two-dimensional (2-D) culture systems cannot provide large numbers of human pluripotent stem cells (hPSCs) and their derivatives that are demanded for commercial and clinical applications in in vitro drug screening, disease modeling, and potentially cell therapy. The technologies that support three-dimensional (3-D) suspension culture, such as a stirred bioreactor, are generally considered as promising approaches to produce the required cells. Recently, suspension bioreactors have also been used to generate mini-brain-like structure from hPSCs for disease modeling, showing the important role of bioreactor in stem cell culture. This chapter describes a detailed culture protocol for neural commitment of hPSCs into neural progenitor cell (NPC) spheres using a spinner bioreactor. The basic steps to prepare hPSCs for bioreactor inoculation are illustrated from cell thawing to cell propagation. The method for generating NPCs from hPSCs in the spinner bioreactor along with the static control is then described. The protocol in this study can be applied to the generation of NPCs from hPSCs for further neural subtype specification, 3-D neural tissue development, or potential preclinical studies or clinical applications in neurological diseases.

Biohydrogen production in a continuous stirred tank bioreactor from synthesis gas by anaerobic photosynthetic bacterium: Rhodopirillum rubrum.

PubMed

Younesi, Habibollah; Najafpour, Ghasem; Ku Ismail, Ku Syahidah; Mohamed, Abdul Rahman; Kamaruddin, Azlina Harun

2008-05-01

Hydrogen may be considered a potential fuel for the future since it is carbon-free and oxidized to water as a combustion product. Bioconversion of synthesis gas (syngas) to hydrogen was demonstrated in continuous stirred tank bioreactor (CSTBR) utilizing acetate as a carbon source. An anaerobic photosynthetic bacterium, Rhodospirillum rubrum catalyzed water-gas shift reaction which was applied for the bioconversion of syngas to hydrogen. The continuous fermentation of syngas in the bioreactor was continuously operated at various gas flow rates and agitation speeds, for the period of two months. The gas flow rates were varied from 5 to 14 ml/min. The agitation speeds were increasingly altered in the range of 150-500 rpm. The pH and temperature of the bioreactor was set at 6.5 and 30 degrees C. The liquid flow rate was kept constant at 0.65 ml/min for the duration of 60 days. The inlet acetate concentration was fed at 4 g/l into the bioreactor. The hydrogen production rate and yield were 16+/-1.1 mmol g(-1)cell h(-1) and 87+/-2.4% at fixed agitation speed of 500 rpm and syngas flow rate of 14 ml/min, respectively. The mass transfer coefficient (KLa) at this condition was approximately 72.8h(-1). This new approach, using a biocatalyst was considered as an alternative method of conventional Fischer-Tropsch synthetic reactions, which were able to convert syngas into hydrogen.

Development of an Autonomous, Dual Chamber Bioreactor for the Growth of 3-Dimensional Epithelial-Stromal Tissues in Microgravity

NASA Technical Reports Server (NTRS)

Patel, Zarana S.; Wettergreen, Matthew A.; Huff, Janice L.

2014-01-01

We are developing a novel, autonomous bioreactor that can provide for the growth and maintenance in microgravity of 3-D organotypic epithelial-stromal cultures that require an air-liquid interface. These complex 3-D tissue models accurately represent the morphological features, differentiation markers, and growth characteristics observed in normal human epithelial tissues, including the skin, esophagus, lung, breast, pancreas, and colon. However, because of their precise and complex culture requirements, including that of an air-liquid interface, these 3-D models have yet to be utilized for life sciences research aboard the International Space Station. The development of a bioreactor for these cultures will provide the capability to perform biological research on the ISS using these realistic, tissue-like human epithelial-stromal cell models and will contribute significantly to advances in fundamental space biology research on questions regarding microgravity effects on normal tissue development, aging, cancer, and other disease processes. It will also allow for the study of how combined stressors, such as microgravity with radiation and nutritional deficiencies, affect multiple biological processes and will provide a platform for conducting countermeasure investigations on the ISS without the use of animal models. The technology will be autonomous and consist of a cell culture chamber that provides for air-liquid, liquid-liquid, and liquid-air exchanges within the chambers while maintaining the growth and development of the biological samples. The bioreactor will support multiple tissue types and its modular design will provide for incorporation of add-on capabilities such as microfluidics drug delivery, media sampling, and in situ biomarker analysis. Preliminary flight testing of the hardware will be conducted on a parabolic platform through NASA's Flight Opportunities Program.

Performance of an under-loaded denitrifying bioreactor with biochar amendment.

PubMed

Bock, Emily M; Coleman, Brady S L; Easton, Zachary M

2018-07-01

Denitrifying bioreactors are recently-established agricultural best management practices with growing acceptance in the US Midwest but less studied in other agriculturally significant regions, such as the US Mid-Atlantic. A bioreactor was installed in the Virginia Coastal Plain to evaluate performance in this geographically novel region facing challenges managing nutrient pollution. The 25.3 m 3 woodchip bed amended with 10% biochar (v/v) intercepted subsurface drainage from 6.5 ha cultivated in soy. Influent and effluent nitrate-nitrogen (NO 3 -N) and total phosphorus (TP) concentrations and flowrate were monitored intensively during the second year of operation. Bed surface fluxes of greenhouse gases (GHGs) nitrous oxide (N 2 O), methane (CH 4 ), and carbon dioxide (CO 2 ) were measured periodically with the closed dynamic chamber technique. The bioreactor did not have a statistically or environmentally significant effect on TP export. Cumulative NO 3 -N removal efficiency (9.5%) and average removal rate (0.56 ± 0.25 g m -3  d -1 ) were low relative to Midwest tile bioreactors, but comparable to installations in the Maryland Coastal Plain. Underperformance was attributed mainly to low NO 3 -N loading (mean 9.4 ± 4.4 kg ha -1 yr -1 ), although intermittent flow, periods of low HRT, and low pH (mean 5.3) also likely contributed. N removal rates were correlated with influent NO 3 -N concentration and temperature, but decreased with hydraulic residence time, indicating that removal was often N-limited. GHG emissions were similar to other bioreactors and constructed wetlands and not considered environmentally concerning. This study suggests that expectations of NO 3 -N removal efficiency developed from bioreactors receiving moderate to high NO 3 -N loading with influent concentrations exceeding 10-20 mg L -1 are unlikely to be met by systems where N-limitation becomes significant. Copyright © 2018 Elsevier Ltd. All rights reserved.

Mathematical modeling and experimental testing of three bioreactor configurations based on windkessel models

PubMed Central

Ruel, Jean; Lachance, Geneviève

2010-01-01

This paper presents an experimental study of three bioreactor configurations. The bioreactor is intended to be used for the development of tissue-engineered heart valve substitutes. Therefore it must be able to reproduce physiological flow and pressure waveforms accurately. A detailed analysis of three bioreactor arrangements is presented using mathematical models based on the windkessel (WK) approach. First, a review of the many applications of this approach in medical studies enhances its fundamental nature and its usefulness. Then the models are developed with reference to the actual components of the bioreactor. This study emphasizes different conflicting issues arising in the design process of a bioreactor for biomedical purposes, where an optimization process is essential to reach a compromise satisfying all conditions. Two important aspects are the need for a simple system providing ease of use and long-term sterility, opposed to the need for an advanced (thus more complex) architecture capable of a more accurate reproduction of the physiological environment. Three classic WK architectures are analyzed, and experimental results enhance the advantages and limitations of each one. PMID:21977286

Reduction of Cr(6+) to Cr(3+) in a packed-bed bioreactor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Turick, C.E.; Apel, W.A.; Camp, C.E.

1997-12-31

Hexavalent chromium, Cr(6{sup +}), is a common and toxic pollutant in soils and waters. Reduction of the mobile Cr(6{sup +}) to the less mobile and less toxic trivalent chromium, Cr(3{sup +}), can be achieved with conventional chemical reduction technologies. Alternatively, Cr(6{sup +}) can be biochemically reduced to Cr(3{sup +}) by anaerobic microbial consortia which appear to use Cr(6{sup +}) as a terminal electron acceptor. A bioprocess for Cr(6{sup +}) reduction has been demonstrated using a packed-bed bioreactor containing ceramic packing, and then compared to a similar bioreactor containing DuPont Bio-Sep beads. An increase in volumetric productivity from 4 mg Cr(6{supmore » +})/L/h to 260 mg Cr(6{sup +})/L/h, probably due to an increase in biomass density, was obtained using Bio-Sep beads. The beads contain internal macropores which were shown by scanning electron microscopy to house dense concentrations of bacteria. Comparisons to conventional Cr(6{sup +}) treatment technologies indicate that a bioprocess has several economic and operational advantages. 7 refs., 4 figs.« less

Uncovering the Potential of Termite Gut Microbiome for Lignocellulose Bioconversion in Anaerobic Batch Bioreactors

PubMed Central

Auer, Lucas; Lazuka, Adèle; Sillam-Dussès, David; Miambi, Edouard; O'Donohue, Michael; Hernandez-Raquet, Guillermina

2017-01-01

Termites are xylophages, being able to digest a wide variety of lignocellulosic biomass including wood with high lignin content. This ability to feed on recalcitrant plant material is the result of complex symbiotic relationships, which involve termite-specific gut microbiomes. Therefore, these represent a potential source of microorganisms for the bioconversion of lignocellulose in bioprocesses targeting the production of carboxylates. In this study, gut microbiomes of four termite species were studied for their capacity to degrade wheat straw and produce carboxylates in controlled bioreactors. All of the gut microbiomes successfully degraded lignocellulose and up to 45% w/w of wheat straw degradation was observed, with the Nasutitermes ephratae gut-microbiome displaying the highest levels of wheat straw degradation, carboxylate production and enzymatic activity. Comparing the 16S rRNA gene diversity of the initial gut inocula to the bacterial communities in lignocellulose degradation bioreactors revealed important changes in community diversity. In particular, taxa such as Spirochaetes and Fibrobacteres that were highly abundant in the initial gut inocula were replaced by Firmicutes and Proteobacteria at the end of incubation in wheat straw bioreactors. Overall, this study demonstrates that termite-gut microbiomes constitute a reservoir of lignocellulose-degrading bacteria that can be harnessed in artificial conditions for biomass conversion processes that lead to the production of useful molecules. PMID:29312279

An economical bioreactor for evaluating biogas potential of particulate biomass.

PubMed

Wilkie, Ann C; Smith, P H; Bordeaux, F M

2004-03-01

An economical bioreactor designed for evaluating the biogas potential of particulate biomass is described. The bioreactor uses a simple stirring apparatus, called the Bordeaux stirrer, to enable gas-tight mixing of fermentation cultures. The apparatus consists of a low-rpm motor connected to a bent steel stir rod, which is placed in a length of flexible plastic tubing inserted through a rubber stopper in a gas-tight manner. This stirrer is suitable for providing intermittent or continuous mixing in bench-scale anaerobic cultures containing particulate biomass. The reactor system may be operated as a batch-fed or semi-continuously fed digester. This communication documents the advantages of the stirring apparatus, describes the details of reactor fabrication and operation, and outlines the type of experimental work for which the bioreactor is suitable.

Anaerobic digestion of citrus waste using two-stage membrane bioreactor

NASA Astrophysics Data System (ADS)

Millati, Ria; Lukitawesa; Dwi Permanasari, Ervina; Wulan Sari, Kartika; Nur Cahyanto, Muhammad; Niklasson, Claes; Taherzadeh, Mohammad J.

2018-03-01

Anaerobic digestion is a promising method to treat citrus waste. However, the presence of limonene in citrus waste inhibits anaerobic digestion process. Limonene is an antimicrobial compound and could inhibit methane forming bacteria that takes a longer time to recover than the injured acid forming bacteria. Hence, volatile fatty acids will be accumulated and methane production will be decreased. One way to solve this problem is by conducting anaerobic digestion process into two stages. The first step is aimed for hydrolysis, acidogenesis, and acetogenesis reactions and the second stage is aimed for methanogenesis reaction. The separation of the system would further allow each stage in their optimum conditions making the process more stable. In this research, anaerobic digestion was carried out in batch operations using 120 ml-glass bottle bioreactors in 2 stages. The first stage was performed in free-cells bioreactor, whereas the second stage was performed in both bioreactor of free cells and membrane bioreactor. In the first stage, the reactor was set into ‘anaerobic’ and ‘semi-aerobic’ conditions to examine the effect of oxygen on facultative anaerobic bacteria in acid production. In the second stage, the protection of membrane towards the cells against limonene was tested. For the first stage, the basal medium was prepared with 1.5 g VS of inoculum and 4.5 g VS of citrus waste. The digestion process was carried out at 55°C for four days. For the second stage, the membrane bioreactor was prepared with 3 g of cells that were encased and sealed in a 3×6 cm2 polyvinylidene fluoride membrane. The medium contained 40 ml basal medium and 10 ml liquid from the first stage. The bioreactors were incubated at 55°C for 2 days under anaerobic condition. The results from the first stage showed that the maximum total sugar under ‘anaerobic’ and ‘semi-aerobic’ conditions was 294.3 g/l and 244.7 g/l, respectively. The corresponding values for total volatile

STATE OF THE PRACTICE FOR BIOREACTOR LANDFILLS - SUMMARY OF USEPA WORKSHOP ON BIOREACTOR LANDFILLS: SUMMARY

EPA Science Inventory

This is a summary of the Workshop on Landfill Bioreactors, held 9/6-7/2000 in Arlington, VA. The purpose of the workshop was to provide a forum to EPA, state and local governments, solid waste industry, and academic research representatives to exchange information and ideas on b...

Performance of high intensity fed-batch mammalian cell cultures in disposable bioreactor systems.

PubMed

Smelko, John Paul; Wiltberger, Kelly Rae; Hickman, Eric Francis; Morris, Beverly Janey; Blackburn, Tobias James; Ryll, Thomas

2011-01-01

The adoption of disposable bioreactor technology as an alternate to traditional nondisposable technology is gaining momentum in the biotechnology industry. Evaluation of current disposable bioreactors systems to sustain high intensity fed-batch mammalian cell culture processes needs to be explored. In this study, an assessment was performed comparing single-use bioreactors (SUBs) systems of 50-, 250-, and 1,000-L operating scales with traditional stainless steel (SS) and glass vessels using four distinct mammalian cell culture processes. This comparison focuses on expansion and production stage performance. The SUB performance was evaluated based on three main areas: operability, process scalability, and process performance. The process performance and operability aspects were assessed over time and product quality performance was compared at the day of harvest. Expansion stage results showed disposable bioreactors mirror traditional bioreactors in terms of cellular growth and metabolism. Set-up and disposal times were dramatically reduced using the SUB systems when compared with traditional systems. Production stage runs for both Chinese hamster ovary and NS0 cell lines in the SUB system were able to model SS bioreactors runs at 100-, 200-, 2,000-, and 15,000-L scales. A single 1,000-L SUB run applying a high intensity fed-batch process was able to generate 7.5 kg of antibody with comparable product quality. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Intelligent Bioreactor Management Information System (IBM-IS) for Mitigation of Greenhouse Gas Emissions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Paul Imhoff; Ramin Yazdani; Don Augenstein

Methane is an important contributor to global warming with a total climate forcing estimated to be close to 20% that of carbon dioxide (CO2) over the past two decades. The largest anthropogenic source of methane in the US is 'conventional' landfills, which account for over 30% of anthropogenic emissions. While controlling greenhouse gas emissions must necessarily focus on large CO2 sources, attention to reducing CH4 emissions from landfills can result in significant reductions in greenhouse gas emissions at low cost. For example, the use of 'controlled' or bioreactor landfilling has been estimated to reduce annual US greenhouse emissions by aboutmore » 15-30 million tons of CO2 carbon (equivalent) at costs between $3-13/ton carbon. In this project we developed or advanced new management approaches, landfill designs, and landfill operating procedures for bioreactor landfills. These advances are needed to address lingering concerns about bioreactor landfills (e.g., efficient collection of increased CH4 generation) in the waste management industry, concerns that hamper bioreactor implementation and the consequent reductions in CH4 emissions. Collectively, the advances described in this report should result in better control of bioreactor landfills and reductions in CH4 emissions. Several advances are important components of an Intelligent Bioreactor Management Information System (IBM-IS).« less

Application of a Parallelizable Perfusion Bioreactor for Physiologic 3D Cell Culture.

PubMed

Egger, Dominik; Spitz, Sarah; Fischer, Monica; Handschuh, Stephan; Glösmann, Martin; Friemert, Benedikt; Egerbacher, Monika; Kasper, Cornelia

2017-01-01

It is crucial but challenging to keep physiologic conditions during the cultivation of 3D cell scaffold constructs for the optimization of 3D cell culture processes. Therefore, we demonstrate the benefits of a recently developed miniaturized perfusion bioreactor together with a specialized incubator system that allows for the cultivation of multiple samples while screening different conditions. Hence, a decellularized bone matrix was tested towards its suitability for 3D osteogenic differentiation under flow perfusion conditions. Subsequently, physiologic shear stress and hydrostatic pressure (HP) conditions were optimized for osteogenic differentiation of human mesenchymal stem cells (MSCs). X-ray computed microtomography and scanning electron microscopy (SEM) revealed a closed cell layer covering the entire matrix. Osteogenic differentiation assessed by alkaline phosphatase activity and SEM was found to be increased in all dynamic conditions. Furthermore, screening of different fluid shear stress (FSS) conditions revealed 1.5 mL/min (equivalent to ∼10 mPa shear stress) to be optimal. However, no distinct effect of HP compared to flow perfusion without HP on osteogenic differentiation was observed. Notably, throughout all experiments, cells cultivated under FSS or HP conditions displayed increased osteogenic differentiation, which underlines the importance of physiologic conditions. In conclusion, the bioreactor system was used for biomaterial testing and to develop and optimize a 3D cell culture process for the osteogenic differentiation of MSCs. Due to its versatility and higher throughput efficiency, we hypothesize that this bioreactor/incubator system will advance the development and optimization of a variety of 3D cell culture processes. © 2017 S. Karger AG, Basel.

Design and Performance of the NASA SCEPTOR Distributed Electric Propulsion Flight Demonstrator

NASA Technical Reports Server (NTRS)

Borer, Nicholas K.; Patterson, Michael D.; Viken, Jeffrey K.; Moore, Mark D.; Clarke, Sean; Redifer, Matthew E.; Christie, Robert J.; Stoll, Alex M.; Dubois, Arthur; Bevirt, JoeBen;

Removal of dichloromethane from waste gas streams using a hybrid bubble column/biofilter bioreactor

PubMed Central

2014-01-01

The performance of a hybrid bubble column/biofilter (HBCB) bioreactor for the removal of dichloromethane (DCM) from waste gas streams was studied in continuous mode for several months. The HBCB bioreactor consisted of two compartments: bubble column bioreactor removing DCM from liquid phase and biofilter removing DCM from gas phase. Effect of inlet DCM concentration on the elimination capacity was examined in the DCM concentration range of 34–359 ppm with loading rates ranged from 2.2 to 22.8 g/m3.h and constant total empty bed retention time (EBRT) of 200 s. In the equal loading rates, the elimination capacity and removal efficiency of the biofilter were higher than the corresponding values of the bubble column bioreactor. The maximum elimination capacity of the HBCB bioreactor was determined to be 15.7 g/m3.h occurred in the highest loading rate of 22.8 g/m3.h with removal efficiency of 69%. The overall mineralization portion of the HBCB bioreactor was in the range of 72-79%. The mixed liquor acidic pH especially below 5.5 inhibited microbial activity and decreased the elimination capacity. Inhibitory effect of high ionic strength was initiated in the mixed liquor electrical conductivity of 12.2 mS/cm. This study indicated that the HBCB bioreactor could benefit from advantages of both bubble column and biofilter reactors and could remove DCM from waste gas streams in a better manner. PMID:24406056

Optimizing T Cell Expansion in a Hollow-Fiber Bioreactor.

PubMed

Nankervis, Brian; Jones, Mark; Vang, Boah; Brent Rice, R; Coeshott, Claire; Beltzer, Jim

2018-01-01

Recent developments in regenerative medicine have precipitated the need to expand gene-modified human T cells to numbers that exceed the capacity of well-plate-based, and flask-based processes. This review discusses the changes in process development that are needed to meet the cell expansion requirements by utilizing hollow-fiber bioreactors . Maintenance of cell proliferation over long periods can become limited by unfilled demands for nutrients and oxygen and by the accumulation of waste products in the local environment. Perfusion feeding, improved gas exchange, and the efficient removal of lactate can increase the yield of T cells from an average of 10.8E +09 to more than 28E +09 in only 10 days. Aggressively feeding cells and actively keeping cells in the bioreactor improves gas exchange and metabolite management over semi-static methods. The ability to remove the environmental constraints that can limit cell expansion by using a two-chamber hollow-fiber bioreactor will be discussed.

Biodegradation of nonylphenol in a continuous packed-bed bioreactor.

PubMed

Soares, Ana; Guieysse, Benoit; Mattiasson, Bo

2003-06-01

A packed bed bioreactor, with 170 ml glass bead carriers and 130 ml medium, was tested for the removal of the endocrine disrupter, nonylphenol, with a Sphingomonas sp. The bioreactor was first continuously fed with medium saturated with nonylphenol in an attempt to simulate groundwater pollution. At best, nonylphenol was degraded by 99.5% at a feeding rate of 69 ml h(-1) and a removal rate of 4.3 mg nonylphenol day(-1), resulting in a 7.5-fold decrease in effluent toxicity according to the Microtox. The bioreactor was then fed with soil leachates at 69 ml h(-1) from artificially contaminated soil (1 g nonylphenol kg(-1) soil) and a real contaminated soil (0.19 g nonylphenol kg(-1) soil). Nonylphenol was always completely removed from the leachates of the two soils. It was removed by 99% from the artificial soil but only 62% from real contaminated soil after 18 and 20 d of treatment, respectively, showing limitation due to nonylphenol adsorption.

High-Lift Propeller System Configuration Selection for NASA's SCEPTOR Distributed Electric Propulsion Flight Demonstrator

NASA Technical Reports Server (NTRS)

Patterson, Michael D.; Derlaga, Joseph M.; Borer, Nicholas K.

2016-01-01

Although the primary function of propellers is typically to produce thrust, aircraft equipped with distributed electric propulsion (DEP) may utilize propellers whose main purpose is to act as a form of high-lift device. These \\high-lift propellers" can be placed upstream of wing such that, when the higher-velocity ow in the propellers' slipstreams interacts with the wing, the lift is increased. This technique is a main design feature of a new NASA advanced design project called Scalable Convergent Electric Propulsion Technology Operations Research (SCEPTOR). The goal of the SCEPTOR project is design, build, and y a DEP aircraft to demonstrate that such an aircraft can be much more ecient than conventional designs. This paper provides details into the high-lift propeller system con guration selection for the SCEPTOR ight demonstrator. The methods used in the high-lift propeller system conceptual design and the tradeo s considered in selecting the number of propellers are discussed.

Application of multivariate analysis and mass transfer principles for refinement of a 3-L bioreactor scale-down model--when shake flasks mimic 15,000-L bioreactors better.

PubMed

Ahuja, Sanjeev; Jain, Shilpa; Ram, Kripa

2015-01-01

Characterization of manufacturing processes is key to understanding the effects of process parameters on process performance and product quality. These studies are generally conducted using small-scale model systems. Because of the importance of the results derived from these studies, the small-scale model should be predictive of large scale. Typically, small-scale bioreactors, which are considered superior to shake flasks in simulating large-scale bioreactors, are used as the scale-down models for characterizing mammalian cell culture processes. In this article, we describe a case study where a cell culture unit operation in bioreactors using one-sided pH control and their satellites (small-scale runs conducted using the same post-inoculation cultures and nutrient feeds) in 3-L bioreactors and shake flasks indicated that shake flasks mimicked the large-scale performance better than 3-L bioreactors. We detail here how multivariate analysis was used to make the pertinent assessment and to generate the hypothesis for refining the existing 3-L scale-down model. Relevant statistical techniques such as principal component analysis, partial least square, orthogonal partial least square, and discriminant analysis were used to identify the outliers and to determine the discriminatory variables responsible for performance differences at different scales. The resulting analysis, in combination with mass transfer principles, led to the hypothesis that observed similarities between 15,000-L and shake flask runs, and differences between 15,000-L and 3-L runs, were due to pCO2 and pH values. This hypothesis was confirmed by changing the aeration strategy at 3-L scale. By reducing the initial sparge rate in 3-L bioreactor, process performance and product quality data moved closer to that of large scale. © 2015 American Institute of Chemical Engineers.

Design of a flow perfusion bioreactor system for bone tissue-engineering applications.

PubMed

Bancroft, Gregory N; Sikavitsas, Vassilios I; Mikos, Antonios G

2003-06-01

Several different bioreactors have been investigated for tissue-engineering applications. Among these bioreactors are the spinner flask and the rotating wall vessel reactor. In addition, a new type of culture system has been developed and investigated, the flow perfusion culture bioreactor. Flow perfusion culture offers several advantages, notably the ability to mitigate both external and internal diffusional limitations as well as to apply mechanical stress to the cultured cells. For such investigation, a flow perfusion culture system was designed and built. This design is the outgrowth of important design requirements and incorporates features crucial to successful experimentation with such a system.

Microalgae-activated sludge treatment of molasses wastewater in sequencing batch photo-bioreactor.

PubMed

Tsioptsias, Costas; Lionta, Gesthimani; Samaras, Petros

2017-05-01

The aim of this work was the examination of the treatment potential of molasses wastewater, by the utilization of activated sludge and microalgae. The systems used included a sequencing batch bioreactor and a similar photo-bioreactor, favoring microalgae growth. The microalgae treatment of molasses wastewater mixture resulted in a considerable reduction in the total nitrogen content. A reduction in the ammonium and nitrate content was observed in the photo-bioreactor, while the effluent's total nitrogen consisted mainly of 50% organic nitrogen. The transformation of the nitrogen forms in the photo-bioreactor was attributed to microalgae activity, resulting in the production of a better quality effluent. Lower COD removal was observed for the photo-bioreactor than the control, which however increased, by the replacement of the anoxic phase by a long aeration period. The mechanism of nitrogen removal included both the denitrification process during the anoxic stage and the microalgae activities, as the replacement of the anoxic stage resulted in low total nitrogen removal capacities. A decrease in the photobioreactor performance was observed after 35 days of operation due to biofilm formation on the light tube surface, while the operation at higher temperature accelerated microalgae growth, resulting thus in the early failure of the photoreactor.

A novel membrane distillation-thermophilic bioreactor system: biological stability and trace organic compound removal.

PubMed

Wijekoon, Kaushalya C; Hai, Faisal I; Kang, Jinguo; Price, William E; Guo, Wenshan; Ngo, Hao H; Cath, Tzahi Y; Nghiem, Long D

2014-05-01

The removal of trace organic compounds (TrOCs) by a novel membrane distillation-thermophilic bioreactor (MDBR) system was examined. Salinity build-up and the thermophilic conditions to some extent adversely impacted the performance of the bioreactor, particularly the removal of total nitrogen and recalcitrant TrOCs. While most TrOCs were well removed by the thermophilic bioreactor, compounds containing electron withdrawing functional groups in their molecular structure were recalcitrant to biological treatment and their removal efficiency by the thermophilic bioreactor was low (0-53%). However, the overall performance of the novel MDBR system with respect to the removal of total organic carbon, total nitrogen, and TrOCs was high and was not significantly affected by the conditions of the bioreactor. All TrOCs investigated here were highly removed (>95%) by the MDBR system. Biodegradation, sludge adsorption, and rejection by MD contribute to the removal of TrOCs by MDBR treatment. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

An Osmotic Membrane Bioreactor-Membrane Distillation System for Simultaneous Wastewater Reuse and Seawater Desalination: Performance and Implications.

PubMed

Luo, Wenhai; Phan, Hop V; Li, Guoxue; Hai, Faisal I; Price, William E; Elimelech, Menachem; Nghiem, Long D

2017-12-19

In this study, we demonstrate the potential of an osmotic membrane bioreactor (OMBR)-membrane distillation (MD) hybrid system for simultaneous wastewater reuse and seawater desalination. A stable OMBR water flux of approximately 6 L m -2 h -1 was achieved when using MD to regenerate the seawater draw solution. Water production by the MD process was higher than that from OMBR to desalinate additional seawater and thus account for draw solute loss due to the reverse salt flux. Amplicon sequencing on the Miseq Illumina platform evidenced bacterial acclimatization to salinity build-up in the bioreactor, though there was a reduction in the bacterial community diversity. In particular, 18 halophilic and halotolerant bacterial genera were identified with notable abundance in the bioreactor. Thus, the effective biological treatment was maintained during OMBR-MD operation. By coupling biological treatment and two high rejection membrane processes, the OMBR-MD hybrid system could effectively remove (>90%) all 30 trace organic contaminants of significant concern investigated here and produce high quality water. Nevertheless, further study is necessary to address MD membrane fouling due to the accumulation of organic matter, particularly protein- and humic-like substances, in seawater draw solution.

Long-term three-dimensional perfusion culture of human adult bone marrow mononuclear cells in bioreactors.

PubMed

Schmelzer, Eva; Finoli, Anthony; Nettleship, Ian; Gerlach, Jörg C

2015-04-01

The construction and long-term maintenance of three-dimensional in vitro bone marrow models is of great interest but still quite challenging. Here we describe the use of a multi-compartment hollow-fiber membrane based three-dimensional perfusion bioreactor for long-term culture of whole human bone marrow mononuclear cells. We also investigated bioreactors with incorporated open-porous foamed hydroxyapatite scaffolds, mimicking the in vivo bone matrix. Cells in bioreactors with and without scaffolds were cultured to 6 weeks and compared to Petri dish controls. Cells were analyzed for gene expression, surface markers by flow cytometry, metabolic activity, hematopoietic potential, viability, and attachment by immunocytochemistry. Cells in bioreactors were metabolic active during long-term culture. The percentages of hematopoietic stem cell and mature endothelial cell fractions were maintained in bioreactors. The expression of most of the analyzed genes stabilized and increased after long-term culture of 6 weeks. Compared to Petri dish culture controls, bioreactor perfusion culture improved in both the short and long-term, the colony formation unit capacity of hematopoietic progenitors. Cells attached to the ample surface area provided by hydroxyapatite scaffolds. The implementation of a hydroxyapatite scaffold did not influence colony formation capacity, percentages of cell type specific fractions, gene expression, cell viability or metabolic turnover when compared to control cells cultured in bioreactors without scaffolds. In conclusion, three-dimensional perfusion bioreactor culture enables long-term maintenance of primary human bone marrow cells, with hydroxyapatite scaffolds providing an in vivo-like scaffold for three-dimensional culture. © 2015 Wiley Periodicals, Inc.

Woodchip bioreactors effectively treat aquaculture effluent

USDA-ARS?s Scientific Manuscript database

Nutrients, in particular nitrogen and phosphorus, can create eutrophication problems in any watershed. Preventing water quality impairment requires controlling nutrients from both point-source and non-point source discharges. Woodchip bioreactors are one relatively new approach that can be utilized ...

A Microfluidic Bioreactor for Toxicity Testing of Stem Cell Derived 3D Cardiac Bodies.

PubMed

Christoffersson, Jonas; Bergström, Gunnar; Schwanke, Kristin; Kempf, Henning; Zweigerdt, Robert; Mandenius, Carl-Fredrik

2016-01-01

Modeling tissues and organs using conventional 2D cell cultures is problematic as the cells rapidly lose their in vivo phenotype. In microfluidic bioreactors the cells reside in microstructures that are continuously perfused with cell culture medium to provide a dynamic environment mimicking the cells natural habitat. These micro scale bioreactors are sometimes referred to as organs-on-chips and are developed in order to improve and extend cell culture experiments. Here, we describe the two manufacturing techniques photolithography and soft lithography that are used in order to easily produce microfluidic bioreactors. The use of these bioreactors is exemplified by a toxicity assessment on 3D clustered human pluripotent stem cells (hPSC)-derived cardiomyocytes by beating frequency imaging.

Integrating human stem cell expansion and neuronal differentiation in bioreactors

PubMed Central

Serra, Margarida; Brito, Catarina; Costa, Eunice M; Sousa, Marcos FQ; Alves, Paula M

2009-01-01

Background Human stem cells are cellular resources with outstanding potential for cell therapy. However, for the fulfillment of this application, major challenges remain to be met. Of paramount importance is the development of robust systems for in vitro stem cell expansion and differentiation. In this work, we successfully developed an efficient scalable bioprocess for the fast production of human neurons. Results The expansion of undifferentiated human embryonal carcinoma stem cells (NTera2/cl.D1 cell line) as 3D-aggregates was firstly optimized in spinner vessel. The media exchange operation mode with an inoculum concentration of 4 × 105 cell/mL was the most efficient strategy tested, with a 4.6-fold increase in cell concentration achieved in 5 days. These results were validated in a bioreactor where similar profile and metabolic performance were obtained. Furthermore, characterization of the expanded population by immunofluorescence microscopy and flow cytometry showed that NT2 cells maintained their stem cell characteristics along the bioreactor culture time. Finally, the neuronal differentiation step was integrated in the bioreactor process, by addition of retinoic acid when cells were in the middle of the exponential phase. Neurosphere composition was monitored and neuronal differentiation efficiency evaluated along the culture time. The results show that, for bioreactor cultures, we were able to increase significantly the neuronal differentiation efficiency by 10-fold while reducing drastically, by 30%, the time required for the differentiation process. Conclusion The culture systems developed herein are robust and represent one-step-forward towards the development of integrated bioprocesses, bridging stem cell expansion and differentiation in fully controlled bioreactors. PMID:19772662

Development of a method for reliable power input measurements in conventional and single‐use stirred bioreactors at laboratory scale

PubMed Central

Werner, Sören; Jossen, Valentin; Kraume, Matthias; Eibl, Dieter

2016-01-01

Power input is an important engineering and scale‐up/down criterion in stirred bioreactors. However, reliably measuring power input in laboratory‐scale systems is still challenging. Even though torque measurements have proven to be suitable in pilot scale systems, sensor accuracy, resolution, and errors from relatively high levels of friction inside bearings can become limiting factors at smaller scales. An experimental setup for power input measurements was developed in this study by focusing on stainless steel and single‐use bioreactors in the single‐digit volume range. The friction losses inside the air bearings were effectively reduced to less than 0.5% of the measurement range of the torque meter. A comparison of dimensionless power numbers determined for a reference Rushton turbine stirrer (N P = 4.17 ± 0.14 for fully turbulent conditions) revealed good agreement with literature data. Hence, the power numbers of several reusable and single‐use bioreactors could be determined over a wide range of Reynolds numbers between 100 and >104. Power numbers of between 0.3 and 4.5 (for Re = 104) were determined for the different systems. The rigid plastic vessels showed similar power characteristics to their reusable counterparts. Thus, it was demonstrated that the torque‐based technique can be used to reliably measure power input in stirred reusable and single‐use bioreactors at the laboratory scale. PMID:28579937

Microgravity

NASA Image and Video Library

1996-01-01

Close-up view of the interior of a NASA Bioreactor shows the plastic plumbing and valves (cylinders at center) to control fluid flow. A fresh nutrient bag is installed at top; a flattened waste bag behind it will fill as the nutrients are consumed during the course of operation. The drive chain and gears for the rotating wall vessel are visible at bottom center center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Bioreactor and process design for biohydrogen production.

PubMed

Show, Kuan-Yeow; Lee, Duu-Jong; Chang, Jo-Shu

2011-09-01

Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. It has the potential for renewable biofuel to replace current hydrogen production which rely heavily on fossil fuels. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of advances in bioreactor and bioprocess design for biohydrogen production. The state-of-the art of biohydrogen production is discussed emphasizing on production pathways, factors affecting biohydrogen production, as well as bioreactor configuration and operation. Challenges and prospects of biohydrogen production are also outlined. Copyright © 2011 Elsevier Ltd. All rights reserved.

Initial results from the Solar Dynamic (SD) Ground Test Demonstration (GTD) project at NASA Lewis

NASA Technical Reports Server (NTRS)

Shaltens, Richard K.; Boyle, Robert V.

1995-01-01

A government/industry team designed, built, and tested a 2 kWe solar dynamic space power system in a large thermal/vacuum facility with a simulated sun at the NASA Lewis Research Center. The Lewis facility provides an accurate simulation of temperatures, high vacuum, and solar flux as encountered in low earth orbit. This paper reviews the goals and status of the Solar Dynamic (SD) Ground Test Demonstration (GTD) program and describes the initial testing, including both operational and performance data. This SD technology has the potential as a future power source for the International Space Station Alpha.

Efficient high-throughput biological process characterization: Definitive screening design with the ambr250 bioreactor system.

PubMed

Tai, Mitchell; Ly, Amanda; Leung, Inne; Nayar, Gautam

2015-01-01

The burgeoning pipeline for new biologic drugs has increased the need for high-throughput process characterization to efficiently use process development resources. Breakthroughs in highly automated and parallelized upstream process development have led to technologies such as the 250-mL automated mini bioreactor (ambr250™) system. Furthermore, developments in modern design of experiments (DoE) have promoted the use of definitive screening design (DSD) as an efficient method to combine factor screening and characterization. Here we utilize the 24-bioreactor ambr250™ system with 10-factor DSD to demonstrate a systematic experimental workflow to efficiently characterize an Escherichia coli (E. coli) fermentation process for recombinant protein production. The generated process model is further validated by laboratory-scale experiments and shows how the strategy is useful for quality by design (QbD) approaches to control strategies for late-stage characterization. © 2015 American Institute of Chemical Engineers.

Microgravity

NASA Image and Video Library

1998-01-01

The heart of the bioreactor is the rotating wall vessel, shown without its support equipment. Volume is about 125 mL. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Computer control of a microgravity mammalian cell bioreactor

NASA Technical Reports Server (NTRS)

Hall, William A.

1987-01-01

The initial steps taken in developing a completely menu driven and totally automated computer control system for a bioreactor are discussed. This bioreactor is an electro-mechanical cell growth system cell requiring vigorous control of slowly changing parameters, many of which are so dynamically interactive that computer control is a necessity. The process computer will have two main functions. First, it will provide continuous environmental control utilizing low signal level transducers as inputs and high powered control devices such as solenoids and motors as outputs. Secondly, it will provide continuous environmental monitoring, including mass data storage and periodic data dumps to a supervisory computer.

Sunlight supply and gas exchange systems in microalgal bioreactor

NASA Technical Reports Server (NTRS)

Mori, K.; Ohya, H.; Matsumoto, K.; Furune, H.

1987-01-01

The bioreactor with sunlight supply system and gas exchange systems presented has proved feasible in ground tests and shows much promise for space use as a closed ecological life support system device. The chief conclusions concerning the specification of total system needed for a life support system for a man in a space station are the following: (1) Sunlight supply system - compactness and low electrical consumption; (2) Bioreactor system - high density and growth rate of chlorella; and (3) Gas exchange system - enough for O2 production and CO2 assimilation.

BIOREACTOR ECONOMICS, SIZE AND TIME OF OPERATION (BEST) COMPUTER SIMULATOR FOR DESIGNING SULFATE-REDUCING BACTERIA FIELD BIOREACTORS

EPA Science Inventory

BEST (bioreactor economics, size and time of operation) is an Excel™ spreadsheet-based model that is used in conjunction with the public domain geochemical modeling software, PHREEQCI. The BEST model is used in the design process of sulfate-reducing bacteria (SRB) field bioreacto...

Example study for granular bioreactor stratification: Three-dimensional evaluation of a sulfate-reducing granular bioreactor

PubMed Central

Hao, Tian-wei; Luo, Jing-hai; Su, Kui-zu; Wei, Li; Mackey, Hamish R.; Chi, Kun; Chen, Guang-Hao

2016-01-01

Recently, sulfate-reducing granular sludge has been developed for application in sulfate-laden water and wastewater treatment. However, little is known about biomass stratification and its effects on the bioprocesses inside the granular bioreactor. A comprehensive investigation followed by a verification trial was therefore conducted in the present work. The investigation focused on the performance of each sludge layer, the internal hydrodynamics and microbial community structures along the height of the reactor. The reactor substratum (the section below baffle 1) was identified as the main acidification zone based on microbial analysis and reactor performance. Two baffle installations increased mixing intensity but at the same time introduced dead zones. Computational fluid dynamics simulation was employed to visualize the internal hydrodynamics. The 16S rRNA gene of the organisms further revealed that more diverse communities of sulfate-reducing bacteria (SRB) and acidogens were detected in the reactor substratum than in the superstratum (the section above baffle 1). The findings of this study shed light on biomass stratification in an SRB granular bioreactor to aid in the design and optimization of such reactors. PMID:27539264

Garver NASA Social

NASA Image and Video Library

2011-05-18

NASA Deputy Administrator Lori Garver, in yellow jacket, stands with participants from the NASA Social underneath the engines of the Saturn V rocket at the Apollo Saturn V visitor center, Thursday, May 18, 2012, at Kennedy Space Center in Cape Canaveral, Fla. About 50 NASA Social followers attended an event as part of activities surrounding the launch of Space Exploration Technologies, or SpaceX, demonstration mission of the company's Falcon 9 rocket to the International Space Station. Photo Credit: (NASA/Paul E. Alers)

Efficacy of Bioremediation of Agricultural Runoff Using Bacterial Communities in Woodchip Bioreactors.

NASA Astrophysics Data System (ADS)

Mortensen, Z. H.; Leandro, M.; Silveus, J. M.

2016-12-01

California's agricultural sector is fundamental in the State's economic growth and is responsible for supplying a large portion of the country's produce. In order to meet the market's demand for crop production the region's agrarian landscape requires an abundance of nutrient rich irrigation. The resultant agricultural effluent is a source of increased nutrient content in California's watershed and groundwater systems, promoting eutrophication and contributing to negative impacts on local ecosystems and human health. Previous studies have examined the denitrification potential of woodchip bioreactors. However, research has been deficient regarding specific variables that may affect the remediation process. To evaluate the efficacy of woodchip bioreactors in remediating waters containing high nitrate concentrations, denitrification rates were examined and parameters such as temperature, laminar flow, and hydraulic residence times were measured to identify potential methods for increasing denitrification efficiency. By measuring the rate of denitrification in a controlled environment where potentially confounding factors can be manipulated, physical components affecting the efficiency of woodchip bioreactors were examined to assess effects. Our research suggests the implementation of woodchip bioreactors to treat agricultural runoff would significantly reduce the concentration of nitrate in agricultural effluent and contribute to the mitigation of negative impacts associated with agricultural irrigation. Future research should focus on the ability of woodchip bioreactors to successfully remediate other agricultural pollutants, such as phosphates and pesticides, to optimize the efficiency of the bioremediation process.

In-depth physiological characterization of primary human hepatocytes in a 3D hollow-fiber bioreactor.

PubMed

Mueller, Daniel; Tascher, Georg; Müller-Vieira, Ursula; Knobeloch, Daniel; Nuessler, Andreas K; Zeilinger, Katrin; Heinzle, Elmar; Noor, Fozia

2011-08-01

As the major research focus is shifting to three-dimensional (3D) cultivation techniques, hollow-fiber bioreactors, allowing the formation of tissue-like structures, show immense potential as they permit controlled in vitro cultivation while supporting the in vivo environment. In this study we carried out a systematic and detailed physiological characterization of human liver cells in a 3D hollow-fiber bioreactor system continuously run for > 2 weeks. Primary human hepatocytes were maintained viable and functional over the whole period of cultivation. Both general cellular functions, e.g. oxygen uptake, amino acid metabolism and substrate consumption, and liver-specific functions, such as drug-metabolizing capacities and the production of liver-specific metabolites were found to be stable for > 2 weeks. As expected, donor-to-donor variability was observed in liver-specific functions, namely urea and albumin production. Moreover, we show the maintenance of primary human hepatocytes in serum-free conditions in this set-up. The stable basal cytochrome P450 activity 3 weeks after isolation of the cells demonstrates the potential of such a system for pharmacological applications. Liver cells in the presented 3D bioreactor system could eventually be used not only for long-term metabolic and toxicity studies but also for chronic repeated dose toxicity assessment. Copyright © 2011 John Wiley & Sons, Ltd.

LANDFILL BIOREACTOR PERFORMANCE, SECOND INTERIM REPORT

EPA Science Inventory

A bioreactor landfill is a landfill that is operated in a manner that is expected to increase the rate and extent of waste decomposition, gas generation, and settlement compared to a traditional landfill. This Second Interim Report was prepared to provide an interpretation of fie...

Comparison of biomass from integrated fixed-film activated sludge (IFAS), moving bed biofilm reactor (MBBR) and membrane bioreactor (MBR) treating recalcitrant organics: Importance of attached biomass.

PubMed

Huang, Chunkai; Shi, Yijing; Xue, Jinkai; Zhang, Yanyan; Gamal El-Din, Mohamed; Liu, Yang

2017-03-15

This study compared microbial characteristics and oil sands process-affected water (OSPW) treatment performance of five types of microbial biomass (MBBR-biofilm, IFAS-biofilm, IFAS-floc, MBR-aerobic-floc, and MBR-anoxic-floc) cultivated from three types of bioreactors (MBBR, IFAS, and MBR) in batch experiments. Chemical oxygen demand (COD), ammonium, acid extractable fraction (AEF), and naphthenic acids (NAs) removals efficiencies were distinctly different between suspended and attached bacterial aggregates and between aerobic and anoxic suspended flocs. MBR-aerobic-floc and MBR-anoxic-floc demonstrated COD removal efficiencies higher than microbial aggregates obtained from MBBR and IFAS, MBBR and IFAS biofilm had higher AEF removal efficiencies than those obtained using flocs. MBBR-biofilm demonstrated the most efficient NAs removal from OSPW. NAs degradation efficiency was highly dependent on the carbon number and NA cyclization number according to UPLC/HRMS analysis. Mono- and di-oxidized NAs were the dominant oxy-NA species in OSPW samples. Microbial analysis with quantitative polymerase chain reaction (q-PCR) indicated that the bacterial 16S rRNA gene abundance was significantly higher in the batch bioreactors with suspended flocs than in those with biofilm, the NSR gene abundance in the MBR-anoxic bioreactor was significantly lower than that in aerobic batch bioreactors, and denitrifiers were more abundant in the suspended phase of the activated sludge flocs. Copyright © 2016 Elsevier B.V. All rights reserved.

Treatment of N-Nitrosodimethylamine (NDMA) in Groundwater Using a Fluidized Bed Bioreactor

DTIC Science & Technology

2014-01-01

Nitrosodimethylamine ( NDMA ) in Groundwater Using a Fluidized Bed Bioreactor Report Documentation Page Form ApprovedOMB No. 0704-0188 Public reporting burden for the...Treatment of N-Nitrosodimethylamine ( NDMA ) in Groundwater Using a Fluidized Bed Bioreactor 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM...21 5.6.1 NDMA and DMN

Design considerations and challenges for mechanical stretch bioreactors in tissue engineering.

PubMed

Lei, Ying; Ferdous, Zannatul

2016-05-01

With the increase in average life expectancy and growing aging population, lack of functional grafts for replacement surgeries has become a severe problem. Engineered tissues are a promising alternative to this problem because they can mimic the physiological function of the native tissues and be cultured on demand. Cyclic stretch is important for developing many engineered tissues such as hearts, heart valves, muscles, and bones. Thus a variety of stretch bioreactors and corresponding scaffolds have been designed and tested to study the underlying mechanism of tissue formation and to optimize the mechanical conditions applied to the engineered tissues. In this review, we look at various designs of stretch bioreactors and common scaffolds and offer insights for future improvements in tissue engineering applications. First, we summarize the requirements and common configuration of stretch bioreactors. Next, we present the features of different actuating and motion transforming systems and their applications. Since most bioreactors must measure detailed distributions of loads and deformations on engineered tissues, techniques with high accuracy, precision, and frequency have been developed. We also cover the key points in designing culture chambers, nutrition exchanging systems, and regimens used for specific tissues. Since scaffolds are essential for providing biophysical microenvironments for residing cells, we discuss materials and technologies used in fabricating scaffolds to mimic anisotropic native tissues, including decellularized tissues, hydrogels, biocompatible polymers, electrospinning, and 3D bioprinting techniques. Finally, we present the potential future directions for improving stretch bioreactors and scaffolds. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:543-553, 2016. © 2016 American Institute of Chemical Engineers.

Simulation of photobioreaction for hydrogen production in membrane bioreactor with an optical fiber

NASA Astrophysics Data System (ADS)

Yang, Yanxia; Li, Jing

2018-05-01

A generalized lattice Boltzmann (LB) model for porous media is adopted to simulate the hydrodynamics and mass transport combined with biodegradation in membrane bioreactor with a circular optical fiber. The LB model is coupled with a multi-block scheme, as well as non-equilibrium extrapolation method for boundary condition treatment. The effect of porosity and permeability (represented by Darcy number Da) of biofilm on flow and concentration fields are investigated. The performance of biodegradation is evaluated by substrate consumption efficiency. Higher porosity and permeability of biofilm facilitate mass transport of substance and enhance the metabolic activity of bacteria in biofilm, which results in the optimal biodegradation performance is obtained under the condition of Da = 0.001 and ɛ =0.3. For further increasing of these parameters, the substrate consumption efficiency decreases due to the inhibition effect of substrate and shorter hydraulic retention time. Furthermore, the LB results coincide with experimental results, demonstrating that the LB model for porous media is available to optimize the membrane bioreactor for efficient biodegradation.

Demonstrating Enabling Technologies for the High-Resolution Imaging Spectrometer of the Next NASA X-ray Astronomy Mission

NASA Astrophysics Data System (ADS)

Kilbourne, Caroline; Adams, J. S.; Bandler, S.; Chervenak, J.; Chiao, M.; Doriese, R.; Eckart, M.; Finkbeiner, F.; Fowler, J. W.; Hilton, G.; Irwin, K.; Kelley, R. L.; Moseley, S. J.; Porter, F. S.; Reintsema, C.; Sadleir, J.; Smith, S. J.; Swetz, D.; Ullom, J.

2014-01-01

NASA/GSFC and NIST-Boulder are collaborating on a program to advance superconducting transition-edge sensor (TES) microcalorimeter technology toward Technology Readiness Level (TRL) 6. The technology development for a TES imaging X-ray microcalorimeter spectrometer (TES microcalorimeter arrays and time-division multiplexed SQUID readout) is now at TRL 4, as evaluated by both NASA and the European Space Agency (ESA) during mission formulation for the International X-ray Observatory (IXO). We will present the status of the development program. The primary goal of the current project is to advance the core X-ray Microcalorimeter Spectrometer (XMS) detector-system technologies to a demonstration of TRL 5 in 2014. Additional objectives are to develop and demonstrate two important related technologies to at least TRL 4: position-sensitive TES devices and code-division multiplexing (CDM). These technologies have the potential to expand significantly the range of possible instrument optimizations; together they allow an expanded focal plane and higher per-pixel count rates without greatly increasing mission resources. The project also includes development of a design concept and critical technologies needed for the thermal, electrical, and mechanical integration of the detector and readout components into the focal-plane assembly. A verified design concept for the packaging of the focal-plane components will be needed for the detector system eventually to advance to TRL 6. Thus, the current project is a targeted development and demonstration program designed to make significant progress in advancing the XMS detector system toward TRL 6, establishing its readiness for a range of possible mission implementations.

Measuring Water in Bioreactor Landfills

NASA Astrophysics Data System (ADS)

Han, B.; Gallagher, V. N.; Imhoff, P. T.; Yazdani, R.; Chiu, P.

2004-12-01

Methane is an important greenhouse gas, and landfills are the largest anthropogenic source in many developed countries. Bioreactor landfills have been proposed as one means of abating greenhouse gas emissions from landfills. Here, the decomposition of organic wastes is enhanced by the controlled addition of water or leachate to maintain optimal conditions for waste decomposition. Greenhouse gas abatement is accomplished by sequestration of photosynthetically derived carbon in wastes, CO2 offsets from energy use of waste derived gas, and mitigation of methane emission from the wastes. Maintaining optimal moisture conditions for waste degradation is perhaps the most important operational parameter in bioreactor landfills. To determine how much water is needed and where to add it, methods are required to measure water within solid waste. However, there is no reliable method that can measure moisture content simply and accurately in the heterogeneous environment typical of landfills. While well drilling and analysis of solid waste samples is sometimes used to determine moisture content, this is an expensive, time-consuming, and destructive procedure. To overcome these problems, a new technology recently developed by hydrologists for measuring water in the vadose zone --- the partitioning tracer test (PTT) --- was evaluated for measuring water in solid waste in a full-scale bioreactor landfill in Yolo County, CA. Two field tests were conducted in different regions of an aerobic bioreactor landfill, with each test measuring water in ≈ 250 ft3 of solid waste. Tracers were injected through existing tubes inserted in the landfill, and tracer breakthrough curves were measured through time from the landfill's gas collection system. Gas samples were analyzed on site using a field-portable gas chromatograph and shipped offsite for more accurate laboratory analysis. In the center of the landfill, PTT measurements indicated that the fraction of the pore space filled with water

NASA Exploration Forum: Human Path to Mars

NASA Image and Video Library

2014-04-29

Randy Lillard, Program Executive for Technology Demonstration Missions of NASA's Space Technology Mission DIrectorate, speaks about the upcoming Low-Density Supersonic Decelerator demonstration during an Exploration Forum showcasing NASA's human exploration path to Mars in the James E. Webb Auditorium at NASA Headquarters on Tuesday, April 29, 2014. Photo Credit: (NASA/Joel Kowsky)

A symbiotic gas exchange between bioreactors enhances microalgal biomass and lipid productivities: taking advantage of complementary nutritional modes.

PubMed

Santos, C A; Ferreira, M E; da Silva, T Lopes; Gouveia, L; Novais, J M; Reis, A

2011-08-01

This paper describes the association of two bioreactors: one photoautotrophic and the other heterotrophic, connected by the gas phase and allowing an exchange of O(2) and CO(2) gases between them, benefiting from a symbiotic effect. The association of two bioreactors was proposed with the aim of improving the microalgae oil productivity for biodiesel production. The outlet gas flow from the autotrophic (O(2) enriched) bioreactor was used as the inlet gas flow for the heterotrophic bioreactor. In parallel, the outlet gas flow from another heterotrophic (CO(2) enriched) bioreactor was used as the inlet gas flow for the autotrophic bioreactor. Aside from using the air supplied from the auto- and hetero-trophic bioreactors as controls, one mixotrophic bioreactor was also studied and used as a model, for its claimed advantage of CO(2) and organic carbon being simultaneously assimilated. The microalga Chlorella protothecoides was chosen as a model due to its ability to grow under different nutritional modes (auto, hetero, and mixotrophic), and its ability to attain a high biomass productivity and lipid content, suitable for biodiesel production. The comparison between heterotrophic, autotrophic, and mixotrophic Chlorella protothecoides growth for lipid production revealed that heterotrophic growth achieved the highest biomass productivity and lipid content (>22%), and furthermore showed that these lipids had the most suitable fatty acid profile in order to produce high quality biodiesel. Both associations showed a higher biomass productivity (10-20%), when comparing the two separately operated bioreactors (controls) which occurred on the fourth day. A more remarkable result would have been seen if in actuality the two bioreactors had been inter-connected in a closed loop. The biomass productivity gain would have been 30% and the lipid productivity gain would have been 100%, as seen by comparing the productivities of the symbiotic assemblage with the sum of the two

Cultivation of Mycobacterium bovis BCG in bioreactors.

PubMed

Dietrich, Guido; Mollenkopf, Hans-Joachim; Weber, Heinz; Knapp, Bernhard; Diehl, Klaus-Dieter; Hess, Jürgen; Blackkolb, Friedrich; Bröker, Michael; Kaufmann, Stefan H E; Hundt, Erika

2002-07-03

The Mycobacterium bovis BCG vaccine for commercial use is classically produced as surface pellicles by culture on synthetic medium. Under these conditions, reproducibility of the cultures and quality assessment are hampered by slow growth of the bacilli, the formation of bacterial aggregates and a high proportion of dead bacilli after processing and final formulation of the vaccine. Here, we established dispersed cultures of M. bovis BCG in synthetic media in small-scale bioreactors. These cultures allow recording and adjusting of culture parameters and give rise to single bacilli with a high degree of live bacteria. In the murine model, bioreactor-grown M. bovis BCG exhibited slightly stronger replication and persistence than the vaccine produced under the classical conditions. The protective efficacy against challenge with M. tuberculosis was identical for both vaccine preparations.

Catalytic bioreactors and methods of using same

DOEpatents

Worden, Robert Mark; Liu, Yangmu Chloe

2017-07-25

Various embodiments provide a bioreactor for producing a bioproduct comprising one or more catalytically active zones located in a housing and adapted to keep two incompatible gaseous reactants separated when in a gas phase, wherein each of the one or more catalytically active zones may comprise a catalytic component retainer and a catalytic component retained within and/or thereon. Each of the catalytically active zones may additionally or alternatively comprise a liquid medium located on either side of the catalytic component retainer. Catalytic component may include a microbial cell culture located within and/or on the catalytic component retainer, a suspended catalytic component suspended in the liquid medium, or a combination thereof. Methods of using various embodiments of the bioreactor to produce a bioproduct, such as isobutanol, are also provided.

Affordable Development and Demonstration of a Small NTR Engine and Stage: A Preliminary NASA, DOE, and Industry Assessment

NASA Technical Reports Server (NTRS)

Borowski, Stanley K.; Sefcik, Robert J.; Fittje, James E.; McCurdy, David R.; Qualls, Arthur L.; Schnitzler, Bruce G.; Werner, James E.; Weitzberg, Abraham; Joyner, Claude R.

2015-01-01

The Nuclear Thermal Rocket (NTR) represents the next evolutionary step in cryogenic liquid rocket engines. Deriving its energy from fission of uranium-235 atoms contained within fuel elements that comprise the engine's reactor core, the NTR can generate high thrust at a specific impulse of approx. 900 seconds or more - twice that of today's best chemical rockets. In FY'11, as part of the AISP project, NASA proposed a Nuclear Thermal Propulsion (NTP) effort that envisioned two key activities - "Foundational Technology Development" followed by system-level "Technology Demonstrations". Five near-term NTP activities identified for Foundational Technology Development became the basis for the NCPS project started in FY'12 and funded by NASA's AES program. During Phase 1 (FY'12-14), the NCPS project was focused on (1) Recapturing fuel processing techniques and fabricating partial length "heritage" fuel elements for the two candidate fuel forms identified by NASA and the DOE - NERVA graphite "composite" and the uranium dioxide (UO2) in tungsten "cermet". The Phase 1 effort also included: (2) Engine Conceptual Design; (3) Mission Analysis and Requirements Definition; (4) Identification of Affordable Options for Ground Testing; and (5) Formulation of an Affordable and Sustainable NTP Development Strategy. During FY'14, a preliminary plan for DDT&E was outlined by GRC, the DOE and industry for NASA HQ that involved significant system-level demonstration projects that included GTD tests at the NNSS, followed by a FTD mission. To reduce development costs, the GTD and FTD tests use a small, low thrust (approx. 7.5 or 16.5 klbf) engine. Both engines use graphite composite fuel and a "common" fuel element design that is scalable to higher thrust (approx. 25 klbf) engines by increasing the number of elements in a larger diameter core that can produce greater thermal power output. To keep the FTD mission cost down, a simple "1-burn" lunar flyby mission was considered along with

Mathematical modelling of cell layer growth in a hollow fibre bioreactor.

PubMed

Chapman, Lloyd A C; Whiteley, Jonathan P; Byrne, Helen M; Waters, Sarah L; Shipley, Rebecca J

2017-04-07

Generating autologous tissue grafts of a clinically useful volume requires efficient and controlled expansion of cell populations harvested from patients. Hollow fibre bioreactors show promise as cell expansion devices, owing to their potential for scale-up. However, further research is required to establish how to specify appropriate hollow fibre bioreactor operating conditions for expanding different cell types. In this study we develop a simple model for the growth of a cell layer seeded on the outer surface of a single fibre in a perfused hollow fibre bioreactor. Nutrient-rich culture medium is pumped through the fibre lumen and leaves the bioreactor via the lumen outlet or passes through the porous fibre walls and cell layer, and out via ports on the outer wall of the extra-capillary space. Stokes and Darcy equations for fluid flow in the fibre lumen, fibre wall, cell layer and extra-capillary space are coupled to reaction-advection-diffusion equations for oxygen and lactate transport through the bioreactor, and to a simple growth law for the evolution of the free boundary of the cell layer. Cells at the free boundary are assumed to proliferate at a rate that increases with the local oxygen concentration, and to die and detach from the layer if the local fluid shear stress or lactate concentration exceed critical thresholds. We use the model to predict operating conditions that maximise the cell layer growth for different cell types. In particular, we predict the optimal flow rate of culture medium into the fibre lumen and fluid pressure imposed at the lumen outlet for cell types with different oxygen demands and fluid shear stress tolerances, and compare the growth of the cell layer when the exit ports on the outside of the bioreactor are open with that when they are closed. Model simulations reveal that increasing the inlet flow rate and outlet fluid pressure increases oxygen delivery to the cell layer and, therefore, the growth rate of cells that are

Stirred suspension bioreactors as a novel method to enrich germ cells from pre-pubertal pig testis.

PubMed

Dores, C; Rancourt, D; Dobrinski, I

2015-05-01

To study spermatogonial stem cells the heterogeneous testicular cell population first needs to be enriched for undifferentiated spermatogonia, which contain the stem cell population. When working with non-rodent models, this step requires working with large numbers of cells. Available cell separation methods rely on differential properties of testicular cell types such as expression of specific cell surface proteins, size, density, or differential adhesion to substrates to separate germ cells from somatic cells. The objective of this study was to develop an approach that allowed germ cell enrichment while providing efficiency of handling large cell numbers. Here, we report the use of stirred suspension bioreactors (SSB) to exploit the adhesion properties of Sertoli cells to enrich cells obtained from pre-pubertal porcine testes for undifferentiated spermatogonia. We also compared the bioreactor approach with an established differential plating method and the combination of both: SSB followed by differential plating. After 66 h of culture, germ cell enrichment in SSBs provided 7.3 ± 1.0-fold (n = 9), differential plating 9.8 ± 2.4-fold (n = 6) and combination of both methods resulted in 9.1 ± 0.3-fold enrichment of germ cells from the initial germ cell population (n = 3). To document functionality of cells recovered from the bioreactor, we demonstrated that cells retained their functional ability to reassemble seminiferous tubules de novo after grafting to mouse hosts and to support spermatogenesis. These results demonstrate that the SSB allows enrichment of germ cells in a controlled and scalable environment providing an efficient method when handling large cell numbers while reducing variability owing to handling. © 2015 American Society of Andrology and European Academy of Andrology.

Effects of Bioreactor Retention Time on Aerobic Microbial Decomposition of CELSS Crop Residues

NASA Technical Reports Server (NTRS)

Strayer, R. F.; Finger, B. W.; Alazraki, M. P.

1997-01-01

The focus of resource recovery research at the KSC-CELSS Breadboard Project has been the evaluation of microbiologically mediated biodegradation of crop residues by manipulation of bioreactor process and environmental variables. We will present results from over 3 years of studies that used laboratory- and breadboard-scale (8 and 120 L working volumes, respectively) aerobic, fed-batch, continuous stirred tank reactors (CSTR) for recovery of carbon and minerals from breadboard grown wheat and white potato residues. The paper will focus on the effects of a key process variable, bioreactor retention time, on response variables indicative of bioreactor performance. The goal is to determine the shortest retention time that is feasible for processing CELSS crop residues, thereby reducing bioreactor volume and weight requirements. Pushing the lower limits of bioreactor retention times will provide useful data for engineers who need to compare biological and physicochemical components. Bioreactor retention times were manipulated to range between 0.25 and 48 days. Results indicate that increases in retention time lead to a 4-fold increase in crop residue biodegradation, as measured by both dry weight losses and CO2 production. A similar overall trend was also observed for crop residue fiber (cellulose and hemicellulose), with a noticeable jump in cellulose degradation between the 5.3 day and 10.7 day retention times. Water-soluble organic compounds (measured as soluble TOC) were appreciably reduced by more than 4-fold at all retention times tested. Results from a study of even shorter retention times (down to 0.25 days), in progress, will also be presented.

A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xu, Qiyong; Tian, Ying; Wang, Shen

2015-07-15

Highlights: • Temporary aeration shortened the initial acid inhibition phase for methanogens. • COD decreased faster in the hybrid bioreactor than that in the anaerobic control. • Methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. • MSW settlement increased with increasing the frequency of intermittent aeration. - Abstract: Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10 months. Themore » hybrid bioreactors were operated in an aerobic–anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia–nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75 d and 60 d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.« less

Microgravity

NASA Image and Video Library

2001-05-31

The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

Microgravity

NASA Image and Video Library

2001-06-01

Cells cultured on Earth (left) typically settle quickly on the bottom of culture vessels due to gravity. In microgravity (right), cells remain suspended and aggregate to form three-dimensional tissue. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

Membrane bioreactors for treating waste streams.

PubMed

Howell, J A; Arnot, T C; Liu, W

2003-03-01

Membrane bioreactors (MBRs) have a number of advantages for treating wastewater containing large quantities of BOD. This paper reviews the inherent advantages of an MBR, which include high potential biomass loadings, lower sludge yields, and retention of specialized organisms that may not settle well in clarifiers. A major problem in effluent treatment occurs when mixed inorganic and organic wastes occur with high concentrations of pollutants. Inorganics that might cause extremes of pH and/or salinity will inhibit microbial growth and only specialized organisms can survive under these conditions. Refractory organics are only biodegraded with difficulty by specialized organisms, which usually do not resist the extreme inorganic environments. The use of membrane bioreactors to help separate the micro-organisms from the inorganic compounds, yet permit the organics to permeate, has been developed in two different designs that are outlined in this paper. The use of membrane contactors in a multimembrane stripping system to treat acidic chlorinated wastes is proposed and discussed.

Transport Advances in Disposable Bioreactors for Liver Tissue Engineering

NASA Astrophysics Data System (ADS)

Catapano, Gerardo; Patzer, John F.; Gerlach, Jörg Christian

Acute liver failure (ALF) is a devastating diagnosis with an overall survival of approximately 60%. Liver transplantation is the therapy of choice for ALF patients but is limited by the scarce availability of donor organs. The prognosis of ALF patients may improve if essential liver functions are restored during liver failure by means of auxiliary methods because liver tissue has the capability to regenerate and heal. Bioartificial liver (BAL) approaches use liver tissue or cells to provide ALF patients with liver-specific metabolism and synthesis products necessary to relieve some of the symptoms and to promote liver tissue regeneration. The most promising BAL treatments are based on the culture of tissue engineered (TE) liver constructs, with mature liver cells or cells that may differentiate into hepatocytes to perform liver-specific functions, in disposable continuous-flow bioreactors. In fact, adult hepatocytes perform all essential liver functions. Clinical evaluations of the proposed BALs show that they are safe but have not clearly proven the efficacy of treatment as compared to standard supportive treatments. Ambiguous clinical results, the time loss of cellular activity during treatment, and the presence of a necrotic core in the cell compartment of many bioreactors suggest that improvement of transport of nutrients, and metabolic wastes and products to or from the cells in the bioreactor is critical for the development of therapeutically effective BALs. In this chapter, advanced strategies that have been proposed over to improve mass transport in the bioreactors at the core of a BAL for the treatment of ALF patients are reviewed.

Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions.

PubMed

Freyer, Nora; Greuel, Selina; Knöspel, Fanny; Gerstmann, Florian; Storch, Lisa; Damm, Georg; Seehofer, Daniel; Foster Harris, Jennifer; Iyer, Rashi; Schubert, Frank; Zeilinger, Katrin

2018-03-15

The accurate prediction of hepatotoxicity demands validated human in vitro models that can close the gap between preclinical animal studies and clinical trials. In this study we investigated the response of primary human liver cells to toxic drug exposure in a perfused microscale 3D liver bioreactor. The cellularized bioreactors were treated with 5, 10, or 30 mM acetaminophen (APAP) used as a reference substance. Lactate production significantly decreased upon treatment with 30 mM APAP ( p < 0.05) and ammonia release significantly increased in bioreactors treated with 10 or 30 mM APAP ( p < 0.0001), indicating APAP-induced dose-dependent toxicity. The release of prostaglandin E2 showed a significant increase at 30 mM APAP ( p < 0.05), suggesting an inflammatory reaction towards enhanced cellular stress. The expression of genes involved in drug metabolism, antioxidant reactions, urea synthesis, and apoptosis was differentially influenced by APAP exposure. Histological examinations revealed that primary human liver cells in untreated control bioreactors were reorganized in tissue-like cell aggregates. These aggregates were partly disintegrated upon APAP treatment, lacking expression of hepatocyte-specific proteins and transporters. In conclusion, our results validate the suitability of the microscale 3D liver bioreactor to detect hepatotoxic effects of drugs in vitro under perfusion conditions.

Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions

PubMed Central

Freyer, Nora; Greuel, Selina; Knöspel, Fanny; Gerstmann, Florian; Storch, Lisa; Damm, Georg; Seehofer, Daniel; Foster Harris, Jennifer; Iyer, Rashi; Schubert, Frank; Zeilinger, Katrin

2018-01-01

The accurate prediction of hepatotoxicity demands validated human in vitro models that can close the gap between preclinical animal studies and clinical trials. In this study we investigated the response of primary human liver cells to toxic drug exposure in a perfused microscale 3D liver bioreactor. The cellularized bioreactors were treated with 5, 10, or 30 mM acetaminophen (APAP) used as a reference substance. Lactate production significantly decreased upon treatment with 30 mM APAP (p < 0.05) and ammonia release significantly increased in bioreactors treated with 10 or 30 mM APAP (p < 0.0001), indicating APAP-induced dose-dependent toxicity. The release of prostaglandin E2 showed a significant increase at 30 mM APAP (p < 0.05), suggesting an inflammatory reaction towards enhanced cellular stress. The expression of genes involved in drug metabolism, antioxidant reactions, urea synthesis, and apoptosis was differentially influenced by APAP exposure. Histological examinations revealed that primary human liver cells in untreated control bioreactors were reorganized in tissue-like cell aggregates. These aggregates were partly disintegrated upon APAP treatment, lacking expression of hepatocyte-specific proteins and transporters. In conclusion, our results validate the suitability of the microscale 3D liver bioreactor to detect hepatotoxic effects of drugs in vitro under perfusion conditions. PMID:29543727

Bioreactor System Using Noninvasive Imaging and Mechanical Stretch for Biomaterial Screening

PubMed Central

Kluge, Jonathan A.; Leisk, Gary G.; Cardwell, Robyn S.; Fernandes, Alexander P.; House, Michael; Ward, Andrew; Dorfmann, A. Luis; Kaplan, David L.

2012-01-01

Screening biomaterial and tissue systems in vitro, for guidance of performance in vivo, remains a major requirement in the field of tissue engineering. It is critical to understand how culture stimulation affects both tissue construct maturation and function, with the goal of eliminating resource-intensive trial-and-error screening and better matching specifications for various in vivo needs. We present a multifunctional and robust bioreactor design that addresses this need. The design enables a range of mechanical inputs, durations, and frequencies to be applied in coordination with noninvasive optical assessments. A variety of biomaterial systems, including micro- and nano-fiber and porous sponge biomaterials, as well as cell-laden tissue engineering constructs were used in validation studies in order to demonstrate the versatility and utility of this new bioreactor design. The silk-based biomaterials highlighted in these studies offered several unique optical signatures for use in label-free nondestructive imaging that allowed for sequential profiling. Both short- and long-term culture studies were conducted to evaluate several practical scenarios of usage: on a short-term basis, we demonstrate that construct cellularity can be monitored by usage of nonpermanent dyes; on a more long-term basis, we show that cell ingrowth can be monitored by GFP-labeling and construct integrity probed with concurrent load/displacement data. The ability to nondestructively track cells, biomaterials, and new matrix formation without harvesting designated samples at each time point will lead to less resource-intensive studies and should enhance our understanding and the discovery of biomaterial designs related to functional tissue engineering. PMID:21298345

Effects of exogenous aerobic bacteria on methane production and biodegradation of municipal solid waste in bioreactors.

PubMed

Ge, Sai; Liu, Lei; Xue, Qiang; Yuan, Zhiming

2016-09-01

Landfill is the most common and efficient ways of municipal solid waste (MSW) disposal and the landfill biogas, mostly methane, is currently utilized to generate electricity and heat. The aim of this work is to study the effects and the role of exogenous aerobic bacteria mixture (EABM) on methane production and biodegradation of MSW in bioreactors. The results showed that the addition of EABM could effectively enhance hydrolysis and acidogenesis processes of MSW degradation, resulting in 63.95% reduction of volatile solid (VS), the highest methane production rate (89.83Lkg(-1) organic matter) ever recorded and a threefold increase in accumulative methane production (362.9L) than the control (127.1L). In addition, it is demonstrated that white-rot fungi (WRF) might further promote the methane production through highly decomposing lignin, but the lower pH value in leachate and longer acidogenesis duration may cause methane production reduced. The data demonstrated that methane production and biodegradation of MSW in bioreactors could be significantly enhanced by EABM via enhanced hydrolysis and acidogenesis processes, and the results are of great economic importance for the future design and management of landfill. Copyright © 2015 Elsevier Ltd. All rights reserved.

Continuous microcarrier-based cell culture in a benchtop microfluidic bioreactor.

PubMed

Abeille, F; Mittler, F; Obeid, P; Huet, M; Kermarrec, F; Dolega, M E; Navarro, F; Pouteau, P; Icard, B; Gidrol, X; Agache, V; Picollet-D'hahan, N

2014-09-21

Microfluidic bioreactors are expected to impact cell therapy and biopharmaceutical production due to their ability to control cellular microenvironments. This work presents a novel approach for continuous cell culture in a microfluidic system. Microcarriers (i.e., microbeads) are used as growth support for anchorage-dependent mammalian cells. This approach eases the manipulation of cells within the system and enables harmless extraction of cells. Moreover, the microbioreactor uses a perfusion function based on the biocompatible integration of a porous membrane to continuously feed the cells. The perfusion rate is optimized through simulations to provide a stable biochemical environment. Thermal management is also addressed to ensure a homogeneous bioreactor temperature. Eventually, incubator-free cell cultures of Drosophila S2 and PC3 cells are achieved over the course of a week using this bioreactor. In future applications, a more efficient alternative to harvesting cells from microcarriers is also anticipated as suggested by our positive results from the microcarrier digestion experiments.

Membrane bioreactors' potential for ethanol and biogas production: a review.

PubMed

Ylitervo, Päivi; Akinbomia, Julius; Taherzadeha, Mohammad J

2013-01-01

Companies developing and producing membranes for different separation purposes, as well as the market for these, have markedly increased in numbers over the last decade. Membrane and separation technology might well contribute to making fuel ethanol and biogas production from lignocellulosic materials more economically viable and productive. Combining biological processes with membrane separation techniques in a membrane bioreactor (MBR) increases cell concentrations extensively in the bioreactor. Such a combination furthermore reduces product inhibition during the biological process, increases product concentration and productivity, and simplifies the separation of product and/or cells. Various MBRs have been studied over the years, where the membrane is either submerged inside the liquid to be filtered, or placed in an external loop outside the bioreactor. All configurations have advantages and drawbacks, as reviewed in this paper. The current review presents an account of the membrane separation technologies, and the research performed on MBRs, focusing on ethanol and biogas production. The advantages and potentials of the technology are elucidated.

Biogas Production from Citrus Waste by Membrane Bioreactor

PubMed Central

Wikandari, Rachma; Millati, Ria; Cahyanto, Muhammad Nur; Taherzadeh, Mohammad J.

2014-01-01

Rapid acidification and inhibition by d-limonene are major challenges of biogas production from citrus waste. As limonene is a hydrophobic chemical, this challenge was encountered using hydrophilic polyvinylidine difluoride (PVDF) membranes in a biogas reactor. The more sensitive methane-producing archaea were encapsulated in the membranes, while freely suspended digesting bacteria were present in the culture as well. In this membrane bioreactor (MBR), the free digesting bacteria digested the citrus wastes and produced soluble compounds, which could pass through the membrane and converted to biogas by the encapsulated cell. As a control experiment, similar digestions were carried out in bioreactors containing the identical amount of just free cells. The experiments were carried out in thermophilic conditions at 55 °C, and hydraulic retention time of 30 days. The organic loading rate (OLR) was started with 0.3 kg VS/m3/day and gradually increased to 3 kg VS/m3/day. The results show that at the highest OLR, MBR was successful to produce methane at 0.33 Nm3/kg VS, while the traditional free cell reactor reduced its methane production to 0.05 Nm3/kg VS. Approximately 73% of the theoretical methane yield was achieved using the membrane bioreactor. PMID:25167328

Living with heterogeneities in bioreactors: understanding the effects of environmental gradients on cells.

PubMed

Lara, Alvaro R; Galindo, Enrique; Ramírez, Octavio T; Palomares, Laura A

2006-11-01

The presence of spatial gradients in fundamental culture parameters, such as dissolved gases, pH, concentration of substrates, and shear rate, among others, is an important problem that frequently occurs in large-scale bioreactors. This problem is caused by a deficient mixing that results from limitations inherent to traditional scale-up methods and practical constraints during large-scale bioreactor design and operation. When cultured in a heterogeneous environment, cells are continuously exposed to fluctuating conditions as they travel through the various zones of a bioreactor. Such fluctuations can affect cell metabolism, yields, and quality of the products of interest. In this review, the theoretical analyses that predict the existence of environmental gradients in bioreactors and their experimental confirmation are reviewed. The origins of gradients in common culture parameters and their effects on various organisms of biotechnological importance are discussed. In particular, studies based on the scale-down methodology, a convenient tool for assessing the effect of environmental heterogeneities, are surveyed.

On-line removal of volatile fatty acids from CELSS anaerobic bioreactor via nanofiltration

NASA Technical Reports Server (NTRS)

Colon, Guillermo

1995-01-01

The CELSS (controlled ecological life support system) resource recovery system, which is a waste processing system, uses aerobic and anaerobic bioreactors to recover plants nutrients and secondary foods from the inedible biomass. The anaerobic degradation of the inedible biomass by means of culture of rumen bacteria,generates organic compounds such as volatile fatty acids (acetic, propionic, butyric, VFA) and ammonia. The presence of VFA in the bioreactor medium at fairly low concentrations decreases the microbial population's metabolic reactions due to end-product inhibition. Technologies to remove VFA continuously from the bioreactor are of high interest. Several candidate technologies were analyzed, such as organic solvent liquid-liquid extraction, adsorption and/or ion exchange, dialysis, electrodialysis, and pressure driven membrane separation processes. The proposed technique for the on-line removal of VFA from the anaerobic bioreactor was a nanofiltration membrane recycle bioreactor. In order to establish the nanofiltration process performance variables before coupling it to the bioreactor, a series of experiments were carried out using a 10,000 MWCO tubular ceramic membrane module. The variables studied were the bioreactor slurry permeation characteristics, such as, the permeate flux, VFA and the nutrient removal rates as a function of applied transmembrane pressure, fluid recirculation velocity, suspended matter concentration, and process operating time. Results indicate that the permeate flux, VFA and nutrients removal rates are directly proportional to the fluid recirculation velocity in the range between 0.6 to 1.0 m/s, applied pressure when these are low than 1.5 bar, and inversely proportional to the total suspended solids concentration in the range between 23,466 to 34,880. At applied pressure higher than 1.5 bar the flux is not more linearly dependent due to concentration polarization and fouling effects over the membrange surface. It was also found

Characterization of extended channel bioreactors for continuous-flow protein production

DOE PAGES

Timm, Andrea C.; Shankles, Peter G.; Foster, Carmen M.; ...

2015-10-02

In this paper, protein based therapeutics are an important class of drugs, used to treat a variety of medical conditions including cancer and autoimmune diseases. Requiring continuous cold storage, and having a limited shelf life, the ability to produce such therapeutics at the point-of-care would open up new opportunities in distributing medicines and treating patients in more remote locations. Here, the authors describe the first steps in the development of a microfluidic platform that can be used for point-of-care protein synthesis. While biologic medicines, including therapeutic proteins, are commonly produced using recombinant deoxyribonucleic acid (DNA) technology in large batch cellmore » cultures, the system developed here utilizes cell-free protein synthesis (CFPS) technology. CFPS is a scalable technology that uses cell extracts containing the biological machinery required for transcription and translation and combines those extracts with DNA, encoding a specific gene, and the additional metabolites required to produce proteins in vitro. While CFPS reactions are typically performed in batch or fed-batch reactions, a well-engineered reaction scheme may improve both the rate of protein production and the economic efficiency of protein synthesis reactions, as well as enable a more streamlined method for subsequent purification of the protein product—all necessary requirements for point-of-care protein synthesis. In this work, the authors describe a new bioreactor design capable of continuous production of protein using cell-free protein synthesis. The bioreactors were designed with three inlets to separate reactive components prior to on-chip mixing, which lead into a long, narrow, serpentine channel. These multiscale, serpentine channel bioreactors were designed to take advantage of microscale diffusion distances across narrow channels in reactors containing enough volume to produce a therapeutic dose of protein, and open the possibility of performing

NASA in the Park, 2018

NASA Image and Video Library

2018-06-20

NASA in the Park on June 16 in Huntsville featured more than 60 exhibits and demonstrations by NASA experts, as well as performances by Marshall musicians, educational opportunities, games and hands-on activities for all ages. Youthful energy is exhibited in the foot rocket demonstration.

Low Boom Flight Demonstrator Briefing

NASA Image and Video Library

2018-04-03

Panelists Peter Iosifidis, program manager, Low-Boom Flight Demonstrator, Lockheed Martin Skunk Works, left, Peter Coen, project manager, Commercial Supersonics Technology Project, Langley Research Center, NASA, center, and Dr. Ed Waggoner, program director, Integrated Aviation Systems Program, NASA, right, are seen behind a model of the Low Boom Flight Demonstrator at a briefing, Tuesday, April 3, 2018 at NASA Headquarters in Washington. This new experimental aircraft will cut cross country travel times in half by flying faster than the speed of sound without creating a sonic boom, enabling travel from New York to Los Angeles in two hours. Photo Credit: (NASA/Aubrey Gemignani)

Low Boom Flight Demonstrator Briefing

NASA Image and Video Library

2018-04-03

Dr. Ed Waggoner, program director, Integrated Aviation Systems Program, NASA, right, speaks on a panel with Peter Iosifidis, program manager, Low-Boom Flight Demonstrator, Lockheed Martin Skunk Works, left, and Peter Coen, project manager, Commercial Supersonics Technology Project, Langley Research Center, NASA, center, at a briefing on the Low Boom Flight Demonstrator, Tuesday, April 3, 2018 at NASA Headquarters in Washington. This new experimental aircraft will cut cross country travel times in half by flying faster than the speed of sound without creating a sonic boom, enabling travel from New York to Los Angeles in two hours. Photo Credit: (NASA/Aubrey Gemignani)

Use of bioreactors in maxillofacial tissue engineering.

PubMed

Depprich, Rita; Handschel, Jörg; Wiesmann, Hans-Peter; Jäsche-Meyer, Janine; Meyer, Ulrich

2008-07-01

Engineering of various oral tissues is a challenging issue in contemporary maxillofacial reconstructive research. In contrast to the classic biomaterial approach, tissue engineering is based on the understanding of cell driven tissue formation, and aims to generate new functional tissues, rather than just to implant non-living space holders. Researchers hope to reach this goal by combining knowledge from biology, physics, materials science, engineering, and medicine in an integrated manner. Several major technical advances have been made in this field during the last decade, and clinical application is at the stage of first clinical trials. A recent limitation of extracorporally engineered cellular substitutes is the problem of growing enlarged tissues ex vivo. One of the main research topics is therefore to scale up artificial tissue constructs for use in extended defect situations. To overcome the monolayer inherent two-dimensional cell assembly, efforts have been made to grow cells in a three-dimensional space. Bioreactors have therefore been in focus for a considerable time to build up enlarged tissues. The shift from the ex vivo approach of cell multiplication to the generation of a real tissue growth is mirrored by the development of bioreactors, enabling scientists to grow more complex tissue constructs. This present review intends to provide an overview of the current state of art in maxillofacial tissue engineering by the use of bioreactors, its limitations and hopes, as well as the future research trends.

Computer modeling movement of biomass in the bioreactors with bubbling mixing

NASA Astrophysics Data System (ADS)

Kuschev, L. A.; Suslov, D. Yu; Alifanova, A. I.

2017-01-01

Recently in the Russian Federation there is an observation of the development of biogas technologies which are used in organic waste conversion of agricultural enterprises, consequently improving the ecological environment. To intensify the process and effective outstanding performance of the acquisition of biogas the application of systems of mixing of bubbling is used. In the case of bubbling mixing of biomass in the bioreactor two-phase portions consisting of biomass and bubbles of gas are formed. The bioreactor computer model with bubble pipeline has been made in a vertical spiral form forming a cone type turned upside down. With the help of computing program of OpenFVM-Flow, an evaluation experiment was conducted to determine the key technological parameters of process of bubbling mixing and to get a visual picture of biomass flows distribution in the bioreactor. For the experimental bioreactor the following equation of V=190 l, speed level, the biomass circulation, and the time of a single cycle of uax =0,029 m/s; QC =0,00087 m3/s, Δtbm .=159 s. In future, we plan to conduct a series of theoretical and experimental researches into the mixing frequency influence on the biogas acquisition process effectiveness.

Microgravity

NASA Image and Video Library

1998-01-01

Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101823 for a version without labels, and No. 0103180 for an operational schematic.

Microgravity

NASA Image and Video Library

1998-01-01

Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101816 for a version without labels, and No. 0103180 for an operational schematic.

Microgravity

NASA Image and Video Library

1998-01-01

Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101825 for a version with major elements labeled, and No. 0103180 for an operational schematic. 0101816

Microgravity

NASA Image and Video Library

1998-01-01

Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators. See No. 0101824 for a version with labels, and No. 0103180 for an operational schematic.

HIGH-PERFORMANCE STEREOSPECIFIC ELASTOMERS FROM BIOREACTORS

USDA-ARS?s Scientific Manuscript database

In 2008, 10 million tons of natural rubber, cis-1,4-polyisoprene, will be produced for commercial use. Every molecule of that product will be produced in a microscopic bioreactor known as the rubber particle. These particles, suspended in an aqueous phase called latex, evolved to produce and store n...

Osteoarthritic human chondrocytes proliferate in 3D co-culture with mesenchymal stem cells in suspension bioreactors.

PubMed

Khurshid, Madiha; Mulet-Sierra, Aillette; Adesida, Adetola; Sen, Arindom

2018-03-01

Osteoarthritis (OA) is a painful disease, characterized by progressive surface erosion of articular cartilage. The use of human articular chondrocytes (hACs) sourced from OA patients has been proposed as a potential therapy for cartilage repair, but this approach is limited by the lack of scalable methods to produce clinically relevant quantities of cartilage-generating cells. Previous studies in static culture have shown that hACs co-cultured with human mesenchymal stem cells (hMSCs) as 3D pellets can upregulate proliferation and generate neocartilage with enhanced functional matrix formation relative to that produced from either cell type alone. However, because static culture flasks are not readily amenable to scale up, scalable suspension bioreactors were investigated to determine if they could support the co-culture of hMSCs and OA hACs under serum-free conditions to facilitate clinical translation of this approach. When hACs and hMSCs (1:3 ratio) were inoculated at 20,000 cells/ml into 125-ml suspension bioreactors and fed weekly, they spontaneously formed 3D aggregates and proliferated, resulting in a 4.75-fold increase over 16 days. Whereas the apparent growth rate was lower than that achieved during co-culture as a 2D monolayer in static culture flasks, bioreactor co-culture as 3D aggregates resulted in a significantly lower collagen I to II mRNA expression ratio and more than double the glycosaminoglycan/DNA content (5.8 vs. 2.5 μg/μg). The proliferation of hMSCs and hACs as 3D aggregates in serum-free suspension culture demonstrates that scalable bioreactors represent an accessible platform capable of supporting the generation of clinical quantities of cells for use in cell-based cartilage repair. Copyright © 2017 John Wiley & Sons, Ltd.

Stoichiometric and kinetic analysis of extreme halophilic Archaea on various substrates in a corrosion resistant bioreactor.

PubMed

Lorantfy, Bettina; Seyer, Bernhard; Herwig, Christoph

2014-01-25

Extreme halophilic Archaea are extremophile species which can thrive in hypersaline environments of up to 3-5 M sodium chloride concentration. Although their ecology and physiology are widely identified on the microbiological level, little emphasis has been laid on quantitative bioprocess development with extreme halophiles. The goal of this study was to establish, on the one hand, a methodological basis for quantitative bioprocess analysis of extreme halophilic Archaea with an extreme halophilic strain as an example. Firstly, as a novel usage, a corrosion resistant bioreactor setup for extreme halophiles has been implemented. Then, paying special attention to total bioprocess quantification approaches, an indirect method for biomass quantification using on-line process signals was introduced. Subsequently, robust quantitative data evaluation methods for halophiles could be developed, providing defined and controlled cultivation conditions in the bioreactor and therefore obtaining suitable quality of on-line as well as off-line datasets. On the other hand, new physiological results of extreme halophiles in bioreactor have also been obtained based on the quantitative methodological tools. For the first time, quantitative data on stoichiometry and kinetics were collected and evaluated on different carbon sources. The results on various substrates were interpreted, with proposed metabolic mechanisms, by linking to the reported primary carbon metabolism of extreme halophilic Archaea. Moreover, results of chemostat cultures demonstrated that extreme halophilic organisms show Monod-kinetics on different sole carbon sources. A diauxic growth pattern was described on a mixture of substrates in batch cultivations. In addition, the methodologies presented here enable one to characterize the utilized strain Haloferax mediterranei (HFX) as a potential new host organism. Thus, this study offers a strong methodological basis as well as a fundamental physiological assessment for

Space Internet-Embedded Web Technologies Demonstration

NASA Technical Reports Server (NTRS)

Foltz, David A.

2001-01-01

The NASA Glenn Research Center recently demonstrated the ability to securely command and control space-based assets by using the Internet and standard Internet Protocols (IP). This is a significant accomplishment because future NASA missions will benefit by using Internet standards-based protocols. The benefits include reduced mission costs and increased mission efficiency. The Internet-Based Space Command and Control System Architecture demonstrated at the NASA Inspection 2000 event proved that this communications architecture is viable for future NASA missions.

Air purification from TCE and PCE contamination in a hybrid bioreactors and biofilter integrated system.

PubMed

Tabernacka, Agnieszka; Zborowska, Ewa; Lebkowska, Maria; Borawski, Maciej

2014-01-15

A two-stage waste air treatment system, consisting of hybrid bioreactors (modified bioscrubbers) and a biofilter, was used to treat waste air containing chlorinated ethenes - trichloroethylene (TCE) and tetrachloroethylene (PCE). The bioreactor was operated with loadings in the range 0.46-5.50gm(-3)h(-1) for TCE and 2.16-9.02gm(-3)h(-1) for PCE. The biofilter loadings were in the range 0.1-0.97gm(-3)h(-1) for TCE and 0.2-2.12gm(-3)h(-1) for PCE. Under low pollutant loadings, the efficiency of TCE elimination was 23-25% in the bioreactor and 54-70% in the biofilter. The efficiency of PCE elimination was 44-60% in the bioreactor and 50-75% in the biofilter. The best results for the bioreactor were observed one week after the pollutant loading was increased. However, the process did not stabilize. In the next seven days contaminant removal efficiency, enzymatic activity and biomass content were all diminished. Copyright © 2013 Elsevier B.V. All rights reserved.

Creating homogenous strain distribution within 3D cell-encapsulated constructs using a simple and cost-effective uniaxial tensile bioreactor: Design and validation study.

PubMed

Subramanian, Gayathri; Elsaadany, Mostafa; Bialorucki, Callan; Yildirim-Ayan, Eda

2017-08-01

Mechanical loading bioreactors capable of applying uniaxial tensile strains are emerging to be a valuable tool to investigate physiologically relevant cellular signaling pathways and biochemical expression. In this study, we have introduced a simple and cost-effective uniaxial tensile strain bioreactor for the application of precise and homogenous uniaxial strains to 3D cell-encapsulated collagen constructs at physiological loading strains (0-12%) and frequencies (0.01-1 Hz). The bioreactor employs silicone-based loading chambers specifically designed to stretch constructs without direct gripping to minimize stress concentration at the ends of the construct and preserve its integrity. The loading chambers are driven by a versatile stepper motor ball-screw actuation system to produce stretching of the constructs. Mechanical characterization of the bioreactor performed through Finite Element Analysis demonstrated that the constructs experienced predominantly uniaxial tensile strain in the longitudinal direction. The strains produced were found to be homogenous over a 15 × 4 × 2 mm region of the construct equivalent to around 60% of the effective region of characterization. The strain values were also shown to be consistent and reproducible during cyclic loading regimes. Biological characterization confirmed the ability of the bioreactor to promote cell viability, proliferation, and matrix organization of cell-encapsulated collagen constructs. This easy-to-use uniaxial tensile strain bioreactor can be employed for studying morphological, structural, and functional responses of cell-embedded matrix systems in response to physiological loading of musculoskeletal tissues. It also holds promise for tissue-engineered strategies that involve delivery of mechanically stimulated cells at the site of injury through a biological carrier to develop a clinically useful therapy for tissue healing. Biotechnol. Bioeng. 2017;114: 1878-1887. © 2017 Wiley Periodicals

Effects of granular activated carbon on methane removal performance and methanotrophic community of a lab-scale bioreactor.