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.

NMR methods for metabolomics of mammalian cell culture bioreactors.

PubMed

Aranibar, Nelly; Reily, Michael D

2014-01-01

Metabolomics has become an important tool for measuring pools of small molecules in mammalian cell cultures expressing therapeutic proteins. NMR spectroscopy has played an important role, largely because it requires minimal sample preparation, does not require chromatographic separation, and is quantitative. The concentrations of large numbers of small molecules in the extracellular media or within the cells themselves can be measured directly on the culture supernatant and on the supernatant of the lysed cells, respectively, and correlated with endpoints such as titer, cell viability, or glycosylation patterns. The observed changes can be used to generate hypotheses by which these parameters can be optimized. This chapter focuses on the sample preparation, data acquisition, and analysis to get the most out of NMR metabolomics data from CHO cell cultures but could easily be extended to other in vitro culture systems.

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.

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.

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).

NASA Bioreactor Schematic

NASA Technical Reports Server (NTRS)

2001-01-01

The schematic depicts the major elements and flow patterns inside the NASA Bioreactor system. Waste and fresh medium are contained in plastic bags placed side-by-side so the waste bag fills as the fresh medium bag is depleted. The compliance vessel contains a bladder to accommodate pressure transients that might damage the system. A peristolic pump moves fluid by squeezing the plastic tubing, thus avoiding potential contamination. 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 Bioreactor

NASA Technical Reports Server (NTRS)

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

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

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.

EMERGING TECHNOLOGY BULLETIN - METHANOTROPHIC BIOREACTOR SYSTEM - BIOTROL, INC.

EPA Science Inventory

BioTrol's Methanotrophic Bioreactor is an above-ground remedial system for water contaminated with halogenated volatile organic compounds, including trichloroethylene (ICE) and related chemicals. Its design features circumvent problems peculiar to treatment of this unique class o...

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.

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.

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...

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.

Denitrification 'Woodchip' Bioreactors for Productive and Sustainable Agricultural Systems

NASA Astrophysics Data System (ADS)

Christianson, L. E.; Summerfelt, S.; Sharrer, K.; Lepine, C.; Helmers, M. J.

2014-12-01

Growing alarm about negative cascading effects of reactive nitrogen in the environment has led to multifaceted efforts to address elevated nitrate-nitrogen levels in water bodies worldwide. The best way to mitigate N-related impacts, such as hypoxic zones and human health concerns, is to convert nitrate to stable, non-reactive dinitrogen gas through the natural process of denitrification. This means denitrification technologies need to be one of our major strategies for tackling the grand challenge of managing human-induced changes to our global nitrogen cycle. While denitrification technologies have historically been focused on wastewater treatment, there is great interest in new lower-tech options for treating effluent and drainage water from one of our largest reactive nitrogen emitters -- agriculture. Denitrification 'woodchip' bioreactors are able to enhance this natural N-conversion via addition of a solid carbon source (e.g., woodchips) and through designs that facilitate development of anoxic conditions required for denitrification. Wood-based denitrification technologies such as woodchip bioreactors and 'sawdust' walls for groundwater have been shown to be effective at reducing nitrate loads in agricultural settings around the world. Designing these systems to be low-maintenance and to avoid removing land from agricultural production has been a primary focus of this "farmer-friendly" technology. This presentation provides a background on woodchip bioreactors including design considerations, N-removal performance, and current research worldwide. Woodchip bioreactors for the agricultural sector are an accessible new option to address society's interest in improving water quality while simultaneously allowing highly productive agricultural systems to continue to provide food in the face of increasing demand, changing global diets, and fluctuating weather.

nmrML: A Community Supported Open Data Standard for the Description, Storage, and Exchange of NMR Data.

PubMed

Schober, Daniel; Jacob, Daniel; Wilson, Michael; Cruz, Joseph A; Marcu, Ana; Grant, Jason R; Moing, Annick; Deborde, Catherine; de Figueiredo, Luis F; Haug, Kenneth; Rocca-Serra, Philippe; Easton, John; Ebbels, Timothy M D; Hao, Jie; Ludwig, Christian; Günther, Ulrich L; Rosato, Antonio; Klein, Matthias S; Lewis, Ian A; Luchinat, Claudio; Jones, Andrew R; Grauslys, Arturas; Larralde, Martin; Yokochi, Masashi; Kobayashi, Naohiro; Porzel, Andrea; Griffin, Julian L; Viant, Mark R; Wishart, David S; Steinbeck, Christoph; Salek, Reza M; Neumann, Steffen

2018-01-02

NMR is a widely used analytical technique with a growing number of repositories available. As a result, demands for a vendor-agnostic, open data format for long-term archiving of NMR data have emerged with the aim to ease and encourage sharing, comparison, and reuse of NMR data. Here we present nmrML, an open XML-based exchange and storage format for NMR spectral data. The nmrML format is intended to be fully compatible with existing NMR data for chemical, biochemical, and metabolomics experiments. nmrML can capture raw NMR data, spectral data acquisition parameters, and where available spectral metadata, such as chemical structures associated with spectral assignments. The nmrML format is compatible with pure-compound NMR data for reference spectral libraries as well as NMR data from complex biomixtures, i.e., metabolomics experiments. To facilitate format conversions, we provide nmrML converters for Bruker, JEOL and Agilent/Varian vendor formats. In addition, easy-to-use Web-based spectral viewing, processing, and spectral assignment tools that read and write nmrML have been developed. Software libraries and Web services for data validation are available for tool developers and end-users. The nmrML format has already been adopted for capturing and disseminating NMR data for small molecules by several open source data processing tools and metabolomics reference spectral libraries, e.g., serving as storage format for the MetaboLights data repository. The nmrML open access data standard has been endorsed by the Metabolomics Standards Initiative (MSI), and we here encourage user participation and feedback to increase usability and make it a successful standard.

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

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

Survival of Acetate in Biodegraded Stream Water DOM: New Insights Based on NMR Spectroscopy

NASA Astrophysics Data System (ADS)

Whitty, S.; Waggoner, D. C.; Bowen, J. C.; Cory, R. M.; Kaplan, L.; Hatcher, P.

2017-12-01

DOM is a complex chemical mixture of high- (HMW) and low-molecular-weight (LMW) organic molecules that serve as the primary energy sources for heterotrophic bacteria in freshwater environments. However, there are still large uncertainties on the composition of DOM that is labile and thus rapidly metabolized. The current thinking is that labile DOM is primarily composed of monosaccharides, amino acids, and other LMW organic acids such as formic, acetic, or propionic among others, although some humic substances also are biologically labile. To test the contribution of LMW organic acids to the labile fraction of DOM, freshwater samples were collected from five streams within the Rio Tempisquito watershed in Costa Rica and subjected to differing degrees of biodegradation using a series of plug-flow bioreactors with residence times ranging from 0.5-150 min. Varying the residence times of bioreactors allows for separation and identification of labile from less labile to more recalcitrant DOM. The stream water fed into the bioreactors had DOC concentrations that ranged from 0.7-1.2 ppm C and the GF/F-filtered stream water as well as the bioreactor effluents were analyzed directly without pre-treatment using proton nuclear magnetic resonance spectroscopy (1H NMR). Small molecules dominated the 1H NMR spectra with the greatest changes, as a function of bioreactor residence time, in the carbohydrate, terminal methyl, and long-chain methylene structures. In contrast, acetate remained relatively constant after 150 min of bioreactor residence time, thus raising the question of why this inherently labile volatile fatty acid was not consumed by stream microbes colonizing bioreactors that otherwise metabolized approximately 35% of the total dissolved organic carbon present in the stream water. We suggest that acetate may resist biodegradation because it is complexed strongly with inorganic cations.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

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...

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

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

NOVEL CERAMIC MEMBRANE BIOREACTOR FOR LOW-FLOW SYSTEMS - PHASE I

EPA Science Inventory

Improved low-flow (50,000 gallons per day) sanitary wastewater treatment systems are needed. CeraMem Corporation's proposed approach includes a membrane bioreactor (MBR) using fully proven biological processes for biological oxygen demand oxidation and (optionally) fo...

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.

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

Microbial Activity In The Peerless Jenny King Sulfate Reducing Bioreactors System

EPA Science Inventory

The Peerless Jenny King treatment system is a series of four sulfate reducing bioreactor cells installed to treat acid mine drainage in the Upper Tenmile Creek Superfund Site located in the Rimini Mining District, near Helena, MT. The system consists of a wetland pretreatment fo...

Phosphorus and water recovery by a novel osmotic membrane bioreactor-reverse osmosis system.

PubMed

Luo, Wenhai; Hai, Faisal I; Price, William E; Guo, Wenshan; Ngo, Hao H; Yamamoto, Kazuo; Nghiem, Long D

2016-01-01

An osmotic membrane bioreactor-reverse osmosis (OMBR-RO) hybrid system integrated with periodic microfiltration (MF) extraction was evaluated for simultaneous phosphorus and clean water recovery from raw sewage. In this hybrid system, the forward osmosis membrane effectively retained inorganic salts and phosphate in the bioreactor, while the MF membrane periodically bled them out for phosphorus recovery with pH adjustment. The RO process was used for draw solute recovery and clean water production. Results show that phosphorus recuperation from the MF permeate was most effective when the solution pH was adjusted to 10, whereby the recovered precipitate contained 15-20% (wt/wt) of phosphorus. Periodic MF extraction also limited salinity build-up in the bioreactor, resulting in a stable biological performance and an increase in water flux during OMBR operation. Despite the build-up of organic matter and ammonia in the draw solution, OMBR-RO allowed for the recovery of high quality reused water. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

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.

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.

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.

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

Application of high-throughput mini-bioreactor system for systematic scale-down modeling, process characterization, and control strategy development.

PubMed

Janakiraman, Vijay; Kwiatkowski, Chris; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

2015-01-01

High-throughput systems and processes have typically been targeted for process development and optimization in the bioprocessing industry. For process characterization, bench scale bioreactors have been the system of choice. Due to the need for performing different process conditions for multiple process parameters, the process characterization studies typically span several months and are considered time and resource intensive. In this study, we have shown the application of a high-throughput mini-bioreactor system viz. the Advanced Microscale Bioreactor (ambr15(TM) ), to perform process characterization in less than a month and develop an input control strategy. As a pre-requisite to process characterization, a scale-down model was first developed in the ambr system (15 mL) using statistical multivariate analysis techniques that showed comparability with both manufacturing scale (15,000 L) and bench scale (5 L). Volumetric sparge rates were matched between ambr and manufacturing scale, and the ambr process matched the pCO2 profiles as well as several other process and product quality parameters. The scale-down model was used to perform the process characterization DoE study and product quality results were generated. Upon comparison with DoE data from the bench scale bioreactors, similar effects of process parameters on process yield and product quality were identified between the two systems. We used the ambr data for setting action limits for the critical controlled parameters (CCPs), which were comparable to those from bench scale bioreactor data. In other words, the current work shows that the ambr15(TM) system is capable of replacing the bench scale bioreactor system for routine process development and process characterization. © 2015 American Institute of Chemical Engineers.

Detecting Microbially Induced Calcite Precipitation in a Model Well-Bore Using Downhole Low-Field NMR

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

Kirkland, Catherine M.; Zanetti, Sam; Grunewald, Elliot

Microbially induced calcite precipitation (MICP) has been widely researched recently due to its relevance for subsurface engineering applications including sealing leakage pathways and permeability modification. These applications of MICP are inherently difficult to monitor nondestructively in time and space. Nuclear magnetic resonance (NMR) can characterize the pore size distributions, porosity, and permeability of subsurface formations. This investigation used a low-field NMR well-logging probe to monitor MICP in a sand-filled bioreactor, measuring NMR signal amplitude and T 2 relaxation over an 8 day experimental period. Following inoculation with the ureolytic bacteria, Sporosarcina pasteurii, and pulsed injections of urea and calcium substrate,more » the NMR measured water content in the reactor decreased to 76% of its initial value. T 2 relaxation distributions bifurcated from a single mode centered about approximately 650 ms into a fast decaying population ( T 2 less than 10 ms) and a larger population with T 2 greater than 1000 ms. The combination of changes in pore volume and surface minerology accounts for the changes in the T 2 distributions. Destructive sampling confirmed final porosity was approximately 88% of the original value. Here, these results indicate the low-field NMR well-logging probe is sensitive to the physical and chemical changes caused by MICP in a laboratory bioreactor.« less

Detecting Microbially Induced Calcite Precipitation in a Model Well-Bore Using Downhole Low-Field NMR

DOE PAGES

Kirkland, Catherine M.; Zanetti, Sam; Grunewald, Elliot; ...

2016-12-20

Microbially induced calcite precipitation (MICP) has been widely researched recently due to its relevance for subsurface engineering applications including sealing leakage pathways and permeability modification. These applications of MICP are inherently difficult to monitor nondestructively in time and space. Nuclear magnetic resonance (NMR) can characterize the pore size distributions, porosity, and permeability of subsurface formations. This investigation used a low-field NMR well-logging probe to monitor MICP in a sand-filled bioreactor, measuring NMR signal amplitude and T 2 relaxation over an 8 day experimental period. Following inoculation with the ureolytic bacteria, Sporosarcina pasteurii, and pulsed injections of urea and calcium substrate,more » the NMR measured water content in the reactor decreased to 76% of its initial value. T 2 relaxation distributions bifurcated from a single mode centered about approximately 650 ms into a fast decaying population ( T 2 less than 10 ms) and a larger population with T 2 greater than 1000 ms. The combination of changes in pore volume and surface minerology accounts for the changes in the T 2 distributions. Destructive sampling confirmed final porosity was approximately 88% of the original value. Here, these results indicate the low-field NMR well-logging probe is sensitive to the physical and chemical changes caused by MICP in a laboratory bioreactor.« less

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.

NASA Bioreactor

NASA Technical Reports Server (NTRS)

1998-01-01

Astronaut John Blaha replaces an exhausted media bag and filled waste bag with fresh bags to continue a bioreactor experiment aboard space station Mir in 1996. 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. This image is from a video downlink. 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).

A bioreactor system for the nitrogen loop in a Controlled Ecological Life Support System

NASA Technical Reports Server (NTRS)

Saulmon, M. M.; Reardon, K. F.; Sadeh, W. Z.

1996-01-01

As space missions become longer in duration, the need to recycle waste into useful compounds rises dramatically. This problem can be addressed by the development of Controlled Ecological Life Support Systems (CELSS) (i.e., Engineered Closed/Controlled Eco-Systems (ECCES)), consisting of human and plant modules. One of the waste streams leaving the human module is urine. In addition to the reclamation of water from urine, recovery of the nitrogen is important because it is an essential nutrient for the plant module. A 3-step biological process for the recycling of nitrogenous waste (urea) is proposed. A packed-bed bioreactor system for this purpose was modeled, and the issues of reaction step segregation, reactor type and volume, support particle size, and pressure drop were addressed. Based on minimization of volume, a bioreactor system consisting of a plug flow immobilized urease reactor, a completely mixed flow immobilized cell reactor to convert ammonia to nitrite, and a plug flow immobilized cell reactor to produce nitrate from nitrite is recommended. It is apparent that this 3-step bioprocess meets the requirements for space applications.

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.

Microbial Activity In The Peerless Jenny King Sulfate Reducing Bioreactor System (Presentation)

EPA Science Inventory

The Peerless Jenny King treatment system is a series of four sulfate reducing bioreactor cells installed to treat acid mine drainage in the Upper Tenmile Creek Superfund Site located in the Rimini Mining District, near Helena MT. The system consists of a wetland pretreatment fol...

Online monitoring of fermentation processes via non-invasive low-field NMR.

PubMed

Kreyenschulte, Dirk; Paciok, Eva; Regestein, Lars; Blümich, Bernhard; Büchs, Jochen

2015-09-01

For the development of biotechnological processes in academia as well as in industry new techniques are required which enable online monitoring for process characterization and control. Nuclear magnetic resonance (NMR) spectroscopy is a promising analytical tool, which has already found broad applications in offline process analysis. The use of online monitoring, however, is oftentimes constrained by high complexity of custom-made NMR bioreactors and considerable costs for high-field NMR instruments (>US$200,000). Therefore, low-field (1) H NMR was investigated in this study in a bypass system for real-time observation of fermentation processes. The new technique was validated with two microbial systems. For the yeast Hansenula polymorpha glycerol consumption could accurately be assessed in spite of the presence of high amounts of complex constituents in the medium. During cultivation of the fungal strain Ustilago maydis, which is accompanied by the formation of several by-products, the concentrations of glucose, itaconic acid, and the relative amount of glycolipids could be quantified. While low-field spectra are characterized by reduced spectral resolution compared to high-field NMR, the compact design combined with the high temporal resolution (15 s-8 min) of spectra acquisition allowed online monitoring of the respective processes. Both applications clearly demonstrate that the investigated technique is well suited for reaction monitoring in opaque media while at the same time it is highly robust and chemically specific. It can thus be concluded that low-field NMR spectroscopy has a great potential for non-invasive online monitoring of biotechnological processes at the research and practical industrial scales. © 2015 Wiley Periodicals, Inc.

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

NASA Technical Reports Server (NTRS)

Sastry, Jagannadha K.

1998-01-01

We conducted a series of experiments using mouse immune-precursor cells, and observed that bioreactor culturing results in the loss of antigen-specific cytotoxic T lymphocyte (CTL) function. The reason for the abrogation of CTL function is microgravity conditions in the bioreactor, but not the antigen per se or its MHC restriction. Similarly, we observed that allostimulation of human PBMC in the bioreactor, but not in the T flask, resulted in the blunting of both allo-CTL function and the NK activity, indicating that the microgravity-associated functional defects are not unique to the mouse system. These results provide further confirmation to the microgravity-associated immune dysfunction, and constitute ground-based confirmatory data for those related to space-travel.

A New Fluidized Bed Bioreactor Based on Diversion-Type Microcapsule Suspension for Bioartificial Liver Systems

PubMed Central

Li, Jianzhou; Yu, Liang; Chen, Ermei; Zhu, Danhua; Zhang, Yimin; Li, LanJuan

2016-01-01

A fluidized bed bioreactor containing encapsulated hepatocytes may be a valuable alternative to a hollow fiber bioreactor for achieving the improved mass transfer and scale-up potential necessary for clinical use. However, a conventional fluidized bed bioreactor (FBB) operating under high perfusion velocity is incapable of providing the desired performance due to the resulting damage to cell-containing microcapsules and large void volume. In this study, we developed a novel diversion-type microcapsule-suspension fluidized bed bioreactor (DMFBB). The void volume in the bioreactor and stability of alginate/chitosan microcapsules were investigated under different flow rates. Cell viability, synthesis and metabolism functions, and expression of metabolizing enzymes at transcriptional levels in an encapsulated hepatocyte line (C3A cells) were determined. The void volume was significantly less in the novel bioreactor than in the conventional FBB. In addition, the microcapsules were less damaged in the DMFBB during the fluidization process as reflected by the results for microcapsule retention rates, swelling, and breakage. Encapsulated C3A cells exhibited greater viability and CYP1A2 and CYP3A4 activity in the DMFBB than in the FBB, although the increases in albumin and urea synthesis were less prominent. The transcription levels of several CYP450-related genes and an albumin-related gene were dramatically greater in cells in the DMFBB than in those in the FBB. Taken together, our results suggest that the DMFBB is a promising alternative for the design of a bioartificial liver system based on a fluidized bed bioreactor with encapsulated hepatocytes for treating patients with acute hepatic failure or other severe liver diseases. PMID:26840840

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.

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.

Large-scale culture of a megakaryocytic progenitor cell line with a single-use bioreactor system.

PubMed

Nurhayati, Retno Wahyu; Ojima, Yoshihiro; Dohda, Takeaki; Kino-Oka, Masahiro

2018-03-01

The increasing application of regenerative medicine has generated a growing demand for stem cells and their derivatives. Single-use bioreactors offer an attractive platform for stem cell expansion owing to their scalability for large-scale production and feasibility of meeting clinical-grade standards. The current work evaluated the capacity of a single-use bioreactor system (1 L working volume) for expanding Meg01 cells, a megakaryocytic (MK) progenitor cell line. Oxygen supply was provided by surface aeration to minimize foaming and orbital shaking was used to promote oxygen transfer. Oxygen transfer rates (k L a) of shaking speeds 50, 100, and 125 rpm were estimated to be 0.39, 1.12, and 10.45 h -1 , respectively. Shaking speed was a critical factor for optimizing cell growth. At 50 rpm, Meg01 cells exhibited restricted growth due to insufficient mixing. A negative effect occurred when the shaking speed was increased to 125 rpm, likely caused by high hydrodynamic shear stress. The bioreactor culture achieved the highest growth profile when shaken at 100 rpm, achieving a total expansion rate up to 5.7-fold with a total cell number of 1.2 ± 0.2 × 10 9 cells L -1 . In addition, cells expanded using the bioreactor system could maintain their potency to differentiate following the MK lineage, as analyzed from specific surface protein and morphological similarity with the cells grown in the conventional culturing system. Our study reports the impact of operational variables such as shaking speed for growth profile and MK differentiation potential of a progenitor cell line in a single-use bioreactor. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:362-369, 2018. © 2017 American Institute of Chemical Engineers.

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

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

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.

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.

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.

Impact of Bioreactor Environment and Recovery Method on the Profile of Bacterial Populations from Water Distribution Systems.

PubMed

Luo, Xia; Jellison, Kristen L; Huynh, Kevin; Widmer, Giovanni

2015-01-01

Multiple rotating annular reactors were seeded with biofilms flushed from water distribution systems to assess (1) whether biofilms grown in bioreactors are representative of biofilms flushed from the water distribution system in terms of bacterial composition and diversity, and (2) whether the biofilm sampling method affects the population profile of the attached bacterial community. Biofilms were grown in bioreactors until thickness stabilized (9 to 11 weeks) and harvested from reactor coupons by sonication, stomaching, bead-beating, and manual scraping. High-throughput sequencing of 16S rRNA amplicons was used to profile bacterial populations from flushed biofilms seeded into bioreactors as well as biofilms recovered from bioreactor coupons by different methods. β diversity between flushed and reactor biofilms was compared to β diversity between (i) biofilms harvested from different reactors and (ii) biofilms harvested by different methods from the same reactor. These analyses showed that average diversity between flushed and bioreactor biofilms was double the diversity between biofilms from different reactors operated in parallel. The diversity between bioreactors was larger than the diversity associated with different biofilm recovery methods. Compared to other experimental variables, the method used to recover biofilms had a negligible impact on the outcome of water biofilm analyses based on 16S amplicon sequencing. Results from this study show that biofilms grown in reactors over 9 to 11 weeks are not representative models of the microbial populations flushed from a distribution system. Furthermore, the bacterial population profile of biofilms grown in replicate reactors from the same flushed water are likely to diverge. However, four common sampling protocols, which differ with respect to disruption of bacterial cells, provide similar information with respect to the 16S rRNA population profile of the biofilm community.

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.

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

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.

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.

Delineating pMDI model reactions with loblolly pine via solution-state NMR spectroscopy. Part 2, Non-catalyzed reactions with the wood cell wall

Treesearch

Daniel J. Yelle; John Ralph; Charles R. Frihart

2011-01-01

Solution-state NMR provides a powerful tool to observe the presence or absence of covalent bonds between wood and adhesives. Finely ground wood can be dissolved in an NMR compatible solvent system containing dimethylsulfoxide-d6 and N-methylimidazole-d6, in which the wood polymers remain largely intact. High-resolution...

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.

Oxygen fires, materials compatibility and system contaminants

NASA Astrophysics Data System (ADS)

Barter, Simon A.; Hillen, Lance W.

An evaluation is made of aircraft fires initiated by oxygen systems, giving attention to such systems' mechanical design and materials-selection factors. While many oxygen compatibility tests are conducted on single components, operational systems become contaminated through frequent use and occasional abuse; it is therefore essential for the designer to have information about the oxygen compatibility of the entire spectrum of potential contaminants and their various combinations. Valve designs are noted to be especially sensitive to system contamination. Gaseous oxygen fire severity is fundamentally determined by the period of oxygen flow. There is a clear need for an automatic shut-off device which would operate when abnormal flow conditions occur.

Influence of dynamic coupled hydro-bio-mechanical processes on response of municipal solid waste and liner system in bioreactor landfills.

PubMed

Reddy, Krishna R; Kumar, Girish; Giri, Rajiv K

2017-05-01

A two-dimensional (2-D) mathematical model is presented to predict the response of municipal solid waste (MSW) of conventional as well as bioreactor landfills undergoing coupled hydro-bio-mechanical processes. The newly developed and validated 2-D coupled mathematical modeling framework combines and simultaneously solves a two-phase flow model based on the unsaturated Richard's equation, a plain-strain formulation of Mohr-Coulomb mechanical model and first-order decay kinetics biodegradation model. The performance of both conventional and bioreactor landfill was investigated holistically, by evaluating the mechanical settlement, extent of waste degradation with subsequent changes in geotechnical properties, landfill slope stability, and in-plane shear behavior (shear stress-displacement) of composite liner system and final cover system. It is concluded that for the given specific conditions considered, bioreactor landfill attained an overall stabilization after a continuous leachate injection of 16years, whereas the stabilization was observed after around 50years of post-closure in conventional landfills, with a total vertical strain of 36% and 37% for bioreactor and conventional landfills, respectively. The significant changes in landfill settlement, the extent of MSW degradation, MSW geotechnical properties, along with their influence on the in-plane shear response of composite liner and final cover system, between the conventional and bioreactor landfills, observed using the mathematical model proposed in this study, corroborates the importance of considering coupled hydro-bio-mechanical processes while designing and predicting the performance of engineered bioreactor landfills. The study underscores the importance of considering the effect of coupled processes while examining the stability and integrity of the liner and cover systems, which form the integral components of a landfill. Moreover, the spatial and temporal variations in the landfill settlement, the

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

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

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

Double Retort System for Materials Compatibility Testing

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

V. Munne; EV Carelli

2006-02-23

With Naval Reactors (NR) approval of the Naval Reactors Prime Contractor Team (NRPCT) recommendation to develop a gas cooled reactor directly coupled to a Brayton power conversion system as the Space Nuclear Power Plant (SNPP) for Project Prometheus (References a and b) there was a need to investigate compatibility between the various materials to be used throughout the SNPP. Of particular interest was the transport of interstitial impurities from the nickel-base superalloys, which were leading candidates for most of the piping and turbine components to the refractory metal alloys planned for use in the reactor core. This kind of contaminationmore » has the potential to affect the lifetime of the core materials. This letter provides technical information regarding the assembly and operation of a double retort materials compatibility testing system and initial experimental results. The use of a double retort system to test materials compatibility through the transfer of impurities from a source to a sink material is described here. The system has independent temperature control for both materials and is far less complex than closed loops. The system is described in detail and the results of three experiments are presented.« less

On the fluid dynamics of a laboratory scale single-use stirred bioreactor.

PubMed

Odeleye, A O O; Marsh, D T J; Osborne, M D; Lye, G J; Micheletti, M

2014-05-24

The commercial success of mammalian cell-derived recombinant proteins has fostered an increase in demand for novel single-use bioreactor (SUB) systems that facilitate greater productivity, increased flexibility and reduced costs (Zhang et al., 2010). These systems exhibit fluid flow regimes unlike those encountered in traditional glass/stainless steel bioreactors because of the way in which they are designed. With such disparate hydrodynamic environments between SUBs currently on the market, traditional scale-up approaches applied to stirred tanks should be revised. One such SUB is the Mobius ® 3 L CellReady, which consists of an upward-pumping marine scoping impeller. This work represents the first experimental study of the flow within the CellReady using a Particle Image Velocimetry (PIV) approach, combined with a biological study into the impact of these fluid dynamic characteristics on cell culture performance. The PIV study was conducted within the actual vessel, rather than using a purpose-built mimic. PIV measurements conveyed a degree of fluid compartmentalisation resulting from the up-pumping impeller. Both impeller tip speed and fluid working volume had an impact upon the fluid velocities and spatial distribution of turbulence within the vessel. Cell cultures were conducted using the GS-CHO cell-line (Lonza) producing an IgG 4 antibody. Disparity in cellular growth and viability throughout the range of operating conditions used (80-350 rpm and 1-2.4 L working volume) was not substantial, although a significant reduction in recombinant protein productivity was found at 350 rpm and 1 L working volume (corresponding to the highest Reynolds number tested in this work). The study shows promise in the use of PIV to improve understanding of the hydrodynamic environment within individual SUBs and allows identification of the critical hydrodynamic parameters under the different flow regimes for compatibility and scalability across the range of bioreactor

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.

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 deg. C (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.

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.

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.

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.

Crosstalk between immune cells and mesenchymal stromal cells in a 3D bioreactor system.

PubMed

Seifert, Martina; Lubitz, Annika; Trommer, Jeanne; Könnig, Darja; Korus, Gabriela; Marx, Uwe; Volk, Hans-Dieter; Duda, Georg; Kasper, Grit; Lehmann, Kerstin; Stolk, Meaghan; Giese, Christoph

2012-11-01

Mesenchymal stromal cells (MSC), known for their high immune modulatory capacity are promising tools for several cell-based therapies. To better mimic the in vivo situation of MSC interactions with immune cells, we applied an artificial lymph node (ALN)-bioreactor culture system combining a miniaturized perfusion bioreactor with a 3D matrix-based cell culture of immune competent cells forming micro-organoids. Rat lymph node cells and allogeneic bone marrow-derived MSCs were seeded in a 20:1 ratio within the agarose matrix of the ALN-reactor. Lymphocytes were pre-incubated with Concanavalin A (ConA) and then co-cultured with MSC in the matrix with additional ConA in the perfusing medium. Live/dead staining showed survival of the co-cultures during the 8-day ALN-reactor run. Paraffin sections of bioreactor matrices were analyzed by proliferating cell nuclear antigen (PCNA)-specific stai-ning to determine MSC proliferation. Immune modulatory capacity was defined by daily analysis of cytokine secretion profiles (TNFa, IFNy, IL-1a, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-12p40/p70, GM-CSF). Cytokine peak secretion at day 2 was significantly inhibited by MSCs for TNFa (96.8 ± 4.8%) and IFNy (88.7 ± 12.0%) in 3D co-cultures. In contrast, other cytokines (IL-1, IL-6, IL-12) were induced. Furthermore, we detected a significantly higher (58.8%) fraction of proliferating MSCs in the presence of immune cells compared to control bioreactors loaded with MSCs only. In the future, this system might be an excellent tool to investigate the mechanisms of MSC-mediated immune modulation during simulated in vivo conditions.

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.

Design and Fabrication of Anatomical Bioreactor Systems Containing Alginate Scaffolds for Cartilage Tissue Engineering

PubMed Central

Gharravi, Anneh Mohammad; Orazizadeh, Mahmoud; Ansari-Asl, Karim; Banoni, Salem; Izadi, Sina; Hashemitabar, Mahmoud

2012-01-01

The aim of the present study was to develop a tissue-engineering approach through alginate gel molding to mimic cartilage tissue in a three-dimensional culture system. The perfusion biomimetic bioreactor was designed to mimic natural joint. The shear stresses exerting on the bioreactor chamber were calculated by Computational Fluid Dynamic (CFD). Several alginate/bovine chondrocyte constructs were prepared, and were cultured in the bioreactor. Histochemical and immunohistochemical staining methods for the presence of glycosaminoglycan(GAG), overall matrix production and type II collagen protein were performed, respectively. The dynamic mechanical device applied a linear mechanical displacement of 2 mm to 10 mm. The CFD modeling indicated peak velocity and maximum wall shear stress were 1.706×10−3 m/s and 0.02407 dyne/cm 2, respectively. Histochemical and immunohistochemical analysis revealed evidence of cartilage-like tissue with lacunas similar to those of natural cartilage and the production of sulfated GAG of matrix by the chondrons, metachromatic territorial matrix-surrounded cells and accumulation of type II collagen around the cells. The present study indicated that when chondrocytes were seeded in alginate hydrogel and cultured in biomimetic cell culture system, cells survived well and secreted newly synthesized matrix led to improvement of chondrogenesis. PMID:23408660

A Novel Bioreactor System for the Assessment of Endothelialization on Deformable Surfaces

PubMed Central

Bachmann, Björn J.; Bernardi, Laura; Loosli, Christian; Marschewski, Julian; Perrini, Michela; Ehrbar, Martin; Ermanni, Paolo; Poulikakos, Dimos; Ferrari, Aldo; Mazza, Edoardo

2016-01-01

The generation of a living protective layer at the luminal surface of cardiovascular devices, composed of an autologous functional endothelium, represents the ideal solution to life-threatening, implant-related complications in cardiovascular patients. The initial evaluation of engineering strategies fostering endothelial cell adhesion and proliferation as well as the long-term tissue homeostasis requires in vitro testing in environmental model systems able to recapitulate the hemodynamic conditions experienced at the blood-to-device interface of implants as well as the substrate deformation. Here, we introduce the design and validation of a novel bioreactor system which enables the long-term conditioning of human endothelial cells interacting with artificial materials under dynamic combinations of flow-generated wall shear stress and wall deformation. The wall shear stress and wall deformation values obtained encompass both the physiological and supraphysiological range. They are determined through separate actuation systems which are controlled based on validated computational models. In addition, we demonstrate the good optical conductivity of the system permitting online monitoring of cell activities through live-cell imaging as well as standard biochemical post-processing. Altogether, the bioreactor system defines an unprecedented testing hub for potential strategies toward the endothelialization or re-endothelialization of target substrates. PMID:27941901

Oxygen Transfer Characteristics of Miniaturized Bioreactor Systems

PubMed Central

Kirk, Timothy V; Szita, Nicolas

2013-01-01

Since their introduction in 2001 miniaturized bioreactor systems have made great advances in function and performance. In this article the dissolved oxygen (DO) transfer performance of submilliliter microbioreactors, and 1–10 mL minibioreactors was examined. Microbioreactors have reached kLa values of 460 h-1, and are offering instrumentation and some functionality comparable to production systems, but at high throughput screening volumes. Minibioreactors, aside from one 1,440 h-1 kLa system, have not offered as high rates of DO transfer, but have demonstrated superior integration with automated fluid handling systems. Microbioreactors have been typically limited to studies with E. coli, while minibioreactors have offered greater versatility in this regard. Further, mathematical relationships confirming the applicability of kLa measurements across all scales have been derived, and alternatives to fluorescence lifetime DO sensors have been evaluated. Finally, the influence on reactor performance of oxygen uptake rate (OUR), and the possibility of its real-time measurement have been explored. Biotechnol. Bioeng. 2013; 110: 1005–1019. © 2012 Wiley Periodicals, Inc. PMID:23280578

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.

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.

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.

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

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.

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.

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

Cardiac tissue engineering using perfusion bioreactor systems

PubMed Central

Radisic, Milica; Marsano, Anna; Maidhof, Robert; Wang, Yadong; Vunjak-Novakovic, Gordana

2009-01-01

This protocol describes tissue engineering of synchronously contractile cardiac constructs by culturing cardiac cell populations on porous scaffolds (in some cases with an array of channels) and bioreactors with perfusion of culture medium (in some cases supplemented with an oxygen carrier). The overall approach is ‘biomimetic’ in nature as it tends to provide in vivo-like oxygen supply to cultured cells and thereby overcome inherent limitations of diffusional transport in conventional culture systems. In order to mimic the capillary network, cells are cultured on channeled elastomer scaffolds that are perfused with culture medium that can contain oxygen carriers. The overall protocol takes 2–4 weeks, including assembly of the perfusion systems, preparation of scaffolds, cell seeding and cultivation, and on-line and end-point assessment methods. This model is well suited for a wide range of cardiac tissue engineering applications, including the use of human stem cells, and high-fidelity models for biological research. PMID:18388955

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.

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

Effect of operating conditions in production of diagnostic Salmonella Enteritidis O-antigen-specific monoclonal antibody in different bioreactor systems.

PubMed

Ayyildiz-Tamis, Duygu; Nalbantsoy, Ayse; Elibol, Murat; Deliloglu-Gurhan, Saime Ismet

2014-01-01

In this study, different cultivation systems such as roller bottles (RB), 5-L stirred-tank bioreactor (STR), and disposable bioreactors were used to cultivate hybridoma for lab-scale production of Salmonella Enteritidis O-antigen-specific monoclonal antibody (MAb). Hybridoma cell line was cultivated in either serum-containing or serum-free medium (SFM) culture conditions. In STR, MAb production scaled up to 4 L, and production capabilities of the cells were also evaluated in different featured production systems. Moreover, the growth parameters of the cells in all production systems such as glucose consumption, lactate and ammonia production, and also MAb productivities were determined. Collected supernatants from the reactors were concentrated by a cross-flow filtration system. In conclusion, cells were not adapted to SFM in RB and STR. Therefore, less MAb titer in both STR and RB systems with SFM was observed compared to the cultures containing fetal bovine serum-supplemented medium. A higher MAb titer was gained in the membrane-aerated system compared to those in STR and RB. Although the highest MAb titer was obtained in the static membrane bioreactor system, the highest productivity was obtained in STR operated in semicontinuous mode with overlay aeration.

Improving the efficiency of quantitative (1)H NMR: an innovative external standard-internal reference approach.

PubMed

Huang, Yande; Su, Bao-Ning; Ye, Qingmei; Palaniswamy, Venkatapuram A; Bolgar, Mark S; Raglione, Thomas V

2014-01-01

The classical internal standard quantitative NMR (qNMR) method determines the purity of an analyte by the determination of a solution containing the analyte and a standard. Therefore, the standard must meet the requirements of chemical compatibility and lack of resonance interference with the analyte as well as a known purity. The identification of such a standard can be time consuming and must be repeated for each analyte. In contrast, the external standard qNMR method utilizes a standard with a known purity to calibrate the NMR instrument. The external standard and the analyte are measured separately, thereby eliminating the matter of chemical compatibility and resonance interference between the standard and the analyte. However, the instrumental factors, including the quality of NMR tubes, must be kept the same. Any deviations will compromise the accuracy of the results. An innovative qNMR method reported herein utilizes an internal reference substance along with an external standard to assume the role of the standard used in the traditional internal standard qNMR method. In this new method, the internal reference substance must only be chemically compatible and be free of resonance-interference with the analyte or external standard whereas the external standard must only be of a known purity. The exact purity or concentration of the internal reference substance is not required as long as the same quantity is added to the external standard and the analyte. The new method reduces the burden of searching for an appropriate standard for each analyte significantly. Therefore the efficiency of the qNMR purity assay increases while the precision of the internal standard method is retained. Copyright © 2013 Elsevier B.V. All rights reserved.

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.

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.

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.

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.

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.

In vitro osteogenesis of human stem cells by using a three-dimensional perfusion bioreactor culture system: a review.

PubMed

Ceccarelli, Gabriele; Bloise, Nora; Vercellino, Marco; Battaglia, Rosalia; Morgante, Lucia; De Angelis, Maria Gabriella Cusella; Imbriani, Marcello; Visai, Livia

2013-04-01

Tissue engineering (by culturing cells on appropriate scaffolds, and using bioreactors to drive the correct bone structure formation) is an attractive alternative to bone grafting or implantation of bone substitutes. Osteogenesis is a biological process that involves many molecular intracellular pathways organized to optimize bone modeling. The use of bioreactor systems and especially the perfusion bioreactor, provides both the technological means to reveal fundamental mechanisms of cell function in a 3D environment, and the potential to improve the quality of engineered tissues. In this mini-review all the characteristics for the production of an appropriate bone construct are analyzed: the stem cell source, scaffolds useful for the seeding of pre-osteoblastic cells and the effects of fluid flow on differentiation and proliferation of bone precursor cells. By automating and standardizing tissue manufacture in controlled closed systems, engineered tissues may reduce the gap between the process of bone formation in vitro and subsequent graft of bone substitutes in vivo.

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

Anchorage-dependent cell cultures are used for the production of viruses, viral vectors, and vaccines, as well as for various cell therapies and tissue engineering applications. Most of these applications currently rely on planar technologies for the generation of biological products. However, as new cell therapy product candidates move from clinical trials towards potential commercialization, planar platforms have proven to be inadequate to meet large-scale manufacturing demand. Therefore, a new scalable platform for culturing anchorage-dependent cells at high cell volumetric concentrations is urgently needed. One promising solution is to grow cells on microcarriers suspended in single-use bioreactors. Toward this goal, a novel bioreactor system utilizing an innovative Vertical-Wheel™ technology was evaluated for its potential to support scalable cell culture process development. Two anchorage-dependent human cell types were used: human lung carcinoma cells (A549 cell line) and human bone marrow-derived mesenchymal stem cells (hMSC). Key hydrodynamic parameters such as power input, mixing time, Kolmogorov length scale, and shear stress were estimated. The performance of Vertical-Wheel bioreactors (PBS-VW) was then evaluated for A549 cell growth and oncolytic adenovirus type 5 production as well as for hMSC expansion. Regarding the first cell model, higher cell growth and number of infectious viruses per cell were achieved when compared with stirred tank (ST) bioreactors. For the hMSC model, although higher percentages of proliferative cells could be reached in the PBS-VW compared with ST bioreactors, no significant differences in the cell volumetric concentration and expansion factor were observed. Noteworthy, the hMSC population generated in the PBS-VW showed a significantly lower percentage of apoptotic cells as well as reduced levels of HLA-DR positive cells. Overall, these results showed that process transfer from ST bioreactor to PBS-VW, and scale-up was

A xenogeneic-free bioreactor system for the clinical-scale expansion of human mesenchymal stem/stromal cells.

PubMed

Dos Santos, Francisco; Campbell, Andrew; Fernandes-Platzgummer, Ana; Andrade, Pedro Z; Gimble, Jeffrey M; Wen, Yuan; Boucher, Shayne; Vemuri, Mohan C; da Silva, Cláudia L; Cabral, Joaquim M S

2014-06-01

The large cell doses (>1 × 10(6)  cells/kg) used in clinical trials with mesenchymal stem/stromal cells (MSC) will require an efficient production process. Moreover, monitoring and control of MSC ex-vivo expansion is critical to provide a safe and reliable cell product. Bioprocess engineering approaches, such as bioreactor technology, offer the adequate tools to develop and optimize a cost-effective culture system for the rapid expansion of human MSC for cellular therapy. Herein, a xenogeneic (xeno)-free microcarrier-based culture system was successfully established for bone marrow (BM) MSC and adipose tissue-derived stem/stromal cell (ASC) cultivation using a 1L-scale controlled stirred-tank bioreactor, allowing the production of (1.1 ± 0.1) × 10(8) and (4.5 ± 0.2) × 10(7) cells for BM MSC and ASC, respectively, after 7 days. Additionally, the effect of different percent air saturation values (%Airsat ) and feeding regime on the proliferation and metabolism of BM MSC was evaluated. No significant differences in cell growth and metabolic patterns were observed under 20% and 9%Airsat . Also, the three different feeding regimes studied-(i) 25% daily medium renewal, (ii) 25% medium renewal every 2 days, and (iii) fed-batch addition of concentrated nutrients and growth factors every 2 days-yielded similar cell numbers, and only slight metabolic differences were observed. Moreover, the immunophenotype (positive for CD73, CD90 and CD105 and negative for CD31, CD80 and HLA-DR) and multilineage differentiative potential of expanded cells were not affected upon bioreactor culture. These results demonstrated the feasibility of expanding human MSC from different sources in a clinically relevant expansion configuration in a controlled microcarrier-based stirred culture system under xeno-free conditions. The further optimization of this bioreactor culture system will represent a crucial step towards an efficient GMP-compliant clinical-scale MSC

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.

On the fluid dynamics of a laboratory scale single-use stirred bioreactor

PubMed Central

Odeleye, A.O.O.; Marsh, D.T.J.; Osborne, M.D.; Lye, G.J.; Micheletti, M.

2014-01-01

The commercial success of mammalian cell-derived recombinant proteins has fostered an increase in demand for novel single-use bioreactor (SUB) systems that facilitate greater productivity, increased flexibility and reduced costs (Zhang et al., 2010). These systems exhibit fluid flow regimes unlike those encountered in traditional glass/stainless steel bioreactors because of the way in which they are designed. With such disparate hydrodynamic environments between SUBs currently on the market, traditional scale-up approaches applied to stirred tanks should be revised. One such SUB is the Mobius® 3 L CellReady, which consists of an upward-pumping marine scoping impeller. This work represents the first experimental study of the flow within the CellReady using a Particle Image Velocimetry (PIV) approach, combined with a biological study into the impact of these fluid dynamic characteristics on cell culture performance. The PIV study was conducted within the actual vessel, rather than using a purpose-built mimic. PIV measurements conveyed a degree of fluid compartmentalisation resulting from the up-pumping impeller. Both impeller tip speed and fluid working volume had an impact upon the fluid velocities and spatial distribution of turbulence within the vessel. Cell cultures were conducted using the GS-CHO cell-line (Lonza) producing an IgG4 antibody. Disparity in cellular growth and viability throughout the range of operating conditions used (80–350 rpm and 1–2.4 L working volume) was not substantial, although a significant reduction in recombinant protein productivity was found at 350 rpm and 1 L working volume (corresponding to the highest Reynolds number tested in this work). The study shows promise in the use of PIV to improve understanding of the hydrodynamic environment within individual SUBs and allows identification of the critical hydrodynamic parameters under the different flow regimes for compatibility and scalability across the range of bioreactor

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.

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.

Trimethylated homoserine functions as the major compatible solute in the globally significant oceanic cyanobacterium Trichodesmium.

PubMed

Pade, Nadin; Michalik, Dirk; Ruth, Wolfgang; Belkin, Natalia; Hess, Wolfgang R; Berman-Frank, Ilana; Hagemann, Martin

2016-11-15

The oceanic N 2 -fixing cyanobacterium Trichodesmium spp. form extensive surface blooms and contribute significantly to marine carbon and nitrogen cycles in the oligotrophic subtropical and tropical oceans. Trichodesmium grows in salinities from 27 to 43 parts per thousand (ppt), yet its salt acclimation strategy remains enigmatic because the genome of Trichodesmium erythraeum strain IMS101 lacks all genes for the biosynthesis of any known compatible solute. Using NMR and liquid chromatography coupled to mass spectroscopy, we identified the main compatible solute in T. erythraeum strain IMS101 as the quaternary ammonium compound N,N,N-trimethyl homoserine (or homoserine betaine) and elucidated its biosynthetic pathway. The identification of this compatible solute explains how Trichodesmium spp. can thrive in the marine system at varying salinities and provides further insight into the diversity of microbial salt acclimation.

Determination of significant factors controlling compatibility of asphalt with synthetic polymers : final report.

DOT National Transportation Integrated Search

1995-06-01

The primary objective of this work is to evaluate several significant factors controlling compatibility between asphalt and synthetic polymers. Employing nuclear magnetic resonance (NMR), Fourier transform infrared (FTIR), differential scanning calor...

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.

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.

In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System.

PubMed

Knöspel, Fanny; Jacobs, Frank; Freyer, Nora; Damm, Georg; De Bondt, An; van den Wyngaert, Ilse; Snoeys, Jan; Monshouwer, Mario; Richter, Marco; Strahl, Nadja; Seehofer, Daniel; Zeilinger, Katrin

2016-04-16

Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR)), while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR) metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.

In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System

PubMed Central

Knöspel, Fanny; Jacobs, Frank; Freyer, Nora; Damm, Georg; De Bondt, An; van den Wyngaert, Ilse; Snoeys, Jan; Monshouwer, Mario; Richter, Marco; Strahl, Nadja; Seehofer, Daniel; Zeilinger, Katrin

2016-01-01

Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR)), while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR) metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro. PMID:27092500

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.

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.

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.

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.

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.

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.

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.

Guide for Oxygen Compatibility Assessments on Oxygen Components and Systems

NASA Technical Reports Server (NTRS)

Rosales, Keisa R.; Shoffstall, Michael S.; Stoltzfus, Joel M.

2007-01-01

Understanding and preventing fire hazards is necessary when designing, maintaining, and operating oxygen systems. Ignition risks can be minimized by controlling heat sources and using materials that will not ignite or will not support burning in the end-use environment. Because certain materials are more susceptible to ignition in oxygen-enriched environments, a compatibility assessment should be performed before the component is introduced into an oxygen system. This document provides an overview of oxygen fire hazards and procedures that are consistent with the latest versions of American Society for Testing and Materials (ASTM) Standards G63 (1999) and G94 (2005) to address fire hazards associated with oxygen systems. This document supersedes the previous edition, NASA Technical Memorandum 104823, Guide for Oxygen Hazards Analyses on Components and Systems (1996). The step-by-step oxygen compatibility assessment method described herein (see Section 4) enables oxygen-system designers, system engineers, and facility managers to determine areas of concern with respect to oxygen compatibility and, ultimately, prevent damage to a system or injury to personnel.

Oxygen Limited Bioreactors System For Nitrogen Removal Using Immobilized Mix Culture

NASA Astrophysics Data System (ADS)

Pathak, B. K.; Sumino, T.; Saiki, Y.; Kazama, F.

2005-12-01

Recently nutrients concentrations especially nitrogen in natural water is alarming in the world wide. Most of the effort is being done on the removal of high concentration of nitrogen especially from the wastewater treatment plants. The removal efficiency is targeted in all considering the effluent discharge standard set by the national environment agency. In many cases, it does not meet the required standard and receiving water is being polluted. Eutrophication in natural water bodies has been reported even if the nitrogen concentration is low and self purification of natural systems itself is not sufficient to remove the nitrogen due to complex phenomenon. In order to recover the pristine water environment, it is very essential to explore bioreactor systems for natural water systems using immobilized mix culture. Microorganism were entrapped in Polyethylene glycol (PEG) prepolymer gel and cut into 3mm cubic immobilized pellets. Four laboratory scale micro bio-reactors having 0.1 L volumes were packed with immobilized pellets with 50% compact ratio. RUN1, RUN2, RUN3 and RUN4 were packed with immobilized pellets from reservoirs sediments, activated sludge (AS), mixed of AS, AG and biodegradable plastic and anaerobic granules (AG) respectively. Water from Shiokawa Reservoirs was feed to all reactors with supplemental ammonia and nitrite nitrogen as specified in the results and discussions. The reactors were operated dark incubated room in continuous flow mode with hydraulic retention time of 12 hours under oxygen limiting condition. Ammonium, nitrate nitrite nitrogen and total organic carbon (TOC) concentrations were measured as described in APWA and AWWA (1998). Laboratory scale four bioreactors containing different combination of immobilized cell were monitored for 218 days. Influent NH4+-N and NO2--N concentration were 2.27±0.43 and 2.05±0.41 mg/l respectively. Average dissolved oxygen concentration and pH in the reactors were 0.40-2.5 mg/l and pH 6

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.

Uhv compatible chopper system

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

Voss, D.E.; Cohen, S.A.

1979-11-01

A time-of-flight system utilizing a novel mechanical chopper has been developed to measure the energy spectrum of neutral deuterium atoms emitted from a tokamak plasma. The chopper system consists of a motor, a magnetically levitated shaft, and a chopper disc. The 20 g disc is rigidly attached to a 159 g shaft assembly and the set is supported against gravity in vacuum by permanent magnets and a stabilizing servo system. All components are uhv compatible to avoid contamination to the tokamak and to the detector. The 25.4 cm OD, .005 cm thick, 301 stainless steel chopper disc has 24 .025more » cm wide slots photoetched at an 11.4 cm radius. An effective aperture time of .55 ..mu..s is achieved during typical steady state operation at 22,500 rpm with a vacuum pressure of 2 x 10/sup -8/ torr.« less

Effects of granular activated carbon on methane removal performance and methanotrophic community of a lab-scale bioreactor.

PubMed

Lee, Eun-Hee; Choi, Sun-Ah; Yi, Taewoo; Kim, Tae Gwan; Lee, Sang-Don; Cho, Kyung-Suk

2015-01-01

Two identical lab-scale bioreactor systems were operated to examine the effects of granular activated carbon (GAC) on methane removal performance and methanotrophic community. Both bioreactor systems removed methane completely at a CH4 loading rate of 71.2 g-CH4·d(-1) for 17 days. However, the methane removal efficiency declined to 88% in the bioreactor without GAC, while the bioreactor amended with GAC showed greater methane removal efficiency of 97% at a CH4 loading rate of 107.5 g-CH4·d(-1). Although quantitative real-time PCR showed that methanotrophic populations were similar levels of 5-10 × 10(8) pmoA gene copy number·VSS(-1) in both systems, GAC addition changed the methanotrophic community composition of the bioreactor systems. Microarray assay revealed that GAC enhanced the type I methanotrophic genera including Methylobacter, Methylomicrobium, and Methylomonas of the system, which suggests that GAC probably provided a favorable environment for type I methanotrophs. These results indicated that GAC is a promising support material in bioreactor systems for CH4 mitigation.

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.

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.

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.

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.

An MRI-compatible hand sensory vibrotactile system.

PubMed

Wang, Fa; Lakshminarayanan, Kishor; Slota, Gregory P; Seo, Na Jin; Webster, John G

2015-01-01

Recently, the application of vibrotactile noise to the wrist or back of the hand has been shown to enhance fingertip tactile sensory perception (Enders et al 2013), supporting the potential for an assistive device worn at the wrist, that generates minute vibrations to help the elderly or patients with sensory deficit. However, knowledge regarding the detailed physiological mechanism behind this sensory improvement in the central nervous system, especially in the human brain, is limited, hindering progress in development and use of such assistive devices. To enable investigation of the impact of vibrotactile noise on sensorimotor brain activity in humans, a magnetic resonance imaging (MRI)-compatible vibrotactile system was developed to provide vibrotactile noise during an MRI of the brain. The vibrotactile system utilizes a remote (outside the MR room) signal amplifier which provides a voltage from -40 to +40 V to drive a 12 mm diameter piezoelectric vibrator (inside the MR room). It is portable and is found to be MRI-compatible which enables its use for neurologic investigation with MRI. The system was also found to induce an improvement in fingertip tactile sensation, consistent with the previous study.

A simple and low-cost permanent magnet system for NMR

NASA Astrophysics Data System (ADS)

Chonlathep, K.; Sakamoto, T.; Sugahara, K.; Kondo, Y.

2017-02-01

We have developed a simple, easy to build, and low-cost magnet system for NMR, of which homogeneity is about 4 ×10-4 at 57 mT, with a pair of two commercially available ferrite magnets. This homogeneity corresponds to about 90 Hz spectral resolution at 2.45 MHz of the hydrogen Larmor frequency. The material cost of this NMR magnet system is little more than 100. The components can be printed by a 3D printer.

Evaluation of micropollutant removal and fouling reduction in a hybrid moving bed biofilm reactor-membrane bioreactor system.

PubMed

Luo, Yunlong; Jiang, Qi; Ngo, Huu H; Nghiem, Long D; Hai, Faisal I; Price, William E; Wang, Jie; Guo, Wenshan

2015-09-01

A hybrid moving bed biofilm reactor-membrane bioreactor (MBBR-MBR) system and a conventional membrane bioreactor (CMBR) were compared in terms of micropollutant removal efficiency and membrane fouling propensity. The results show that the hybrid MBBR-MBR system could effectively remove most of the selected micropollutants. By contrast, the CMBR system showed lower removals of ketoprofen, carbamazepine, primidone, bisphenol A and estriol by 16.2%, 30.1%, 31.9%, 34.5%, and 39.9%, respectively. Mass balance calculations suggest that biological degradation was the primary removal mechanism in the MBBR-MBR system. During operation, the MBBR-MBR system exhibited significantly slower fouling development as compared to the CMBR system, which could be ascribed to the wide disparity in the soluble microbial products (SMP) levels between MBBR-MBR (4.02-6.32 mg/L) and CMBR (21.78 and 33.04 mg/L). It is evident that adding an MBBR process prior to MBR treatment can not only enhance micropollutant elimination but also mitigate membrane fouling. Copyright © 2015 Elsevier Ltd. All rights reserved.

Multimission Modular Spacecraft Ground Support Software System (MMS/GSSS) state-of-the-art computer systems/ compatibility study

NASA Technical Reports Server (NTRS)

1980-01-01

The compatibility of the Multimission Modular Spacecraft (MMS) Ground Support Software System (GSSS), currently operational on a ModComp IV/35, with the VAX 11/780 system is discussed. The compatibility is examined in various key areas of the GSSS through the results of in depth testing performed on the VAX 11/780 and ModComp IV/35 systems. The compatibility of the GSSS with the ModComp CLASSIC is presented based upon projections from ModComp supplied literature.

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.

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.

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.

Large-scale production of lentiviral vector in a closed system hollow fiber bioreactor

PubMed Central

Sheu, Jonathan; Beltzer, Jim; Fury, Brian; Wilczek, Katarzyna; Tobin, Steve; Falconer, Danny; Nolta, Jan; Bauer, Gerhard

2015-01-01

Lentiviral vectors are widely used in the field of gene therapy as an effective method for permanent gene delivery. While current methods of producing small scale vector batches for research purposes depend largely on culture flasks, the emergence and popularity of lentiviral vectors in translational, preclinical and clinical research has demanded their production on a much larger scale, a task that can be difficult to manage with the numbers of producer cell culture flasks required for large volumes of vector. To generate a large scale, partially closed system method for the manufacturing of clinical grade lentiviral vector suitable for the generation of induced pluripotent stem cells (iPSCs), we developed a method employing a hollow fiber bioreactor traditionally used for cell expansion. We have demonstrated the growth, transfection, and vector-producing capability of 293T producer cells in this system. Vector particle RNA titers after subsequent vector concentration yielded values comparable to lentiviral iPSC induction vector batches produced using traditional culture methods in 225 cm2 flasks (T225s) and in 10-layer cell factories (CF10s), while yielding a volume nearly 145 times larger than the yield from a T225 flask and nearly three times larger than the yield from a CF10. Employing a closed system hollow fiber bioreactor for vector production offers the possibility of manufacturing large quantities of gene therapy vector while minimizing reagent usage, equipment footprint, and open system manipulation. PMID:26151065

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...

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.

Comparison of strategies for the isolation of PCR-compatible, genomic DNA from a municipal biogas plants.

PubMed

Weiss, Agnes; Jérôme, Valérie; Freitag, Ruth

2007-06-15

The goal of the project was the extraction of PCR-compatible genomic DNA representative of the entire microbial community from municipal biogas plant samples (mash, bioreactor content, process water, liquid fertilizer). For the initial isolation of representative DNA from the respective lysates, methods were used that employed adsorption, extraction, or precipitation to specifically enrich the DNA. Since no dedicated method for biogas plant samples was available, preference was given to kits/methods suited to samples that resembled either the bioreactor feed, e.g. foodstuffs, or those intended for environmental samples including wastewater. None of the methods succeeded in preparing DNA that was directly PCR-compatible. Instead the DNA was found to still contain considerable amounts of difficult-to-remove enzyme inhibitors (presumably humic acids) that hindered the PCR reaction. Based on the isolation method that gave the highest yield/purity for all sample types, subsequent purification was attempted by agarose gel electrophoresis followed by electroelution, spermine precipitation, or dialysis through nitrocellulose membrane. A combination of phenol/chloroform extraction followed by purification via dialysis constituted the most efficient sample treatment. When such DNA preparations were diluted 1:100 they did no longer inhibit PCR reactions, while they still contained sufficient genomic DNA to allow specific amplification of specific target sequences.

A simple and low-cost permanent magnet system for NMR.

PubMed

Chonlathep, K; Sakamoto, T; Sugahara, K; Kondo, Y

2017-02-01

We have developed a simple, easy to build, and low-cost magnet system for NMR, of which homogeneity is about 4×10 -4 at 57mT, with a pair of two commercially available ferrite magnets. This homogeneity corresponds to about 90Hz spectral resolution at 2.45MHz of the hydrogen Larmor frequency. The material cost of this NMR magnet system is little more than $100. The components can be printed by a 3D printer. Copyright © 2016 Elsevier Inc. All rights reserved.

Development of a Hybrid EPR/NMR Coimaging System

PubMed Central

Samouilov, Alexandre; Caia, George L.; Kesselring, Eric; Petryakov, Sergey; Wasowicz, Tomasz; Zweier, Jay L.

2010-01-01

Electron paramagnetic resonance imaging (EPRI) is a powerful technique that enables spatial mapping of free radicals or other paramagnetic compounds; however, it does not in itself provide anatomic visualization of the body. Proton magnetic resonance imaging (MRI) is well suited to provide anatomical visualization. A hybrid EPR/NMR coimaging instrument was constructed that utilizes the complementary capabilities of both techniques, superimposing EPR and proton-MR images to provide the distribution of paramagnetic species in the body. A common magnet and field gradient system is utilized along with a dual EPR and proton-NMR resonator assembly, enabling coimaging without the need to move the sample. EPRI is performed at ~1.2 GHz/~40 mT and proton MRI is performed at 16.18 MHz/~380 mT; hence the method is suitable for whole-body coimaging of living mice. The gradient system used is calibrated and controlled in such a manner that the spatial geometry of the two acquired images is matched, enabling their superposition without additional postprocessing or marker registration. The performance of the system was tested in a series of phantoms and in vivo applications by mapping the location of a paramagnetic probe in the gastrointestinal (GI) tract of mice. This hybrid EPR/NMR coimaging instrument enables imaging of paramagnetic molecules along with their anatomic localization in the body. PMID:17659621

MR-Compatible Integrated Eye Tracking System

DTIC Science & Technology

2016-03-10

SECURITY CLASSIFICATION OF: This instrumentation grant was used to purchase state-of-the-art, high-resolution video eye tracker that can be used to...P.O. Box 12211 Research Triangle Park, NC 27709-2211 video eye tracking, eye movments, visual search; camouflage-breaking REPORT DOCUMENTATION PAGE...Report: MR-Compatible Integrated Eye Tracking System Report Title This instrumentation grant was used to purchase state-of-the-art, high-resolution video

Squid-based CW NMR system for measuring the magnetization of helium-3 films

NASA Astrophysics Data System (ADS)

White, Kevin Spencer

This thesis describes the design and construction of a SQUID-based CW NMR system together with its application in a study of the two dimensional magnetism of 3He. 3He provides an exemplary system for the study of two-dimensional magnetism. Two-dimensional 3He films of varying coverages may be formed by plating 3He on relatively uniform two-dimensional substrates, such as GTA Grafoil and ZYX graphite substrates. At coverages above approximately 20 atoms/nm. 2 on these substrates, the second layer of 3He exhibits a strong ferromagnetic ordering tendency. The ferromagnetic ordering presents as a rapid onset of measured magnetization that becomes independent of the applied magnetic field as film temperatures approach 1 mK. Very low applied magnetic fields are used to probe the ferromagnetic ordering in order to minimize masking of the measured magnetization and to stay within the available bandwidth of the SQUID. Commensurate with the ferromagnetic ordering, the NMR linewidth increases dramatically at these coverages and temperatures. An increasing linewidth equates to a short decay time with respect to pulsed NMR probing of the two-dimensional 3He magnetization. The decay times at these coverages and temperatures become so short that they fall below the minimum recovery time necessary for a SQUID-based pulsed NMR system to recover from the relatively large tipping pulse and acquire meaningful data. To address this problem, we have designed a SQUID-based CW NMR system to leverage as much of an already-existing pulsed NMR system as possible but allow accurate measurement of the rapid onset of ferromagnetic ordering of the 3He films below the approximate 1 mK temperature limit of the pulsed NMR system.

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

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

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.

[Compatibility of different quality control systems].

PubMed

Invernizzi, Enrico

2002-01-01

Management of the good laboratory practice (GLP) quality system presupposes its linking to a basic recognized and approved quality system, from which it can draw on management procedures common to all quality systems, such as the ISO 9000 set of norms. A quality system organized in this way can also be integrated with other dedicated quality systems, or parts of them, to obtain principles or management procedures for specific topics. The aim of this organization is to set up a reliable, recognized quality system compatible with the principles of GLP and other quality management systems, which provides users with a simplified set of easily accessible management tools and answers. The organization of this quality system is set out in the quality assurance programme, which is actually the document in which the test facility incorporates the GLP principles into its own quality organization.

NASA Bioreactor tissue culture

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 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.

Building Systems Information Clearinghouse Special Report Number One: Manufacturers Compatibility Study.

ERIC Educational Resources Information Center

Boice, John R., Ed.

BSIC has selected data for inclusion, and a method of presentation that-- (1) provides preliminary data, in comparable form, about all relevant systems building products, (2) surveys within the limits imposed, the problems of compatibility between subsystem components and to identify components which are compatible with one another, (3) identifies…

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.

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

Evolution of uni- and bifactorial sexual compatibility systems in fungi

PubMed Central

Nieuwenhuis, B P S; Billiard, S; Vuilleumier, S; Petit, E; Hood, M E; Giraud, T

2013-01-01

Mating systems, that is, whether organisms give rise to progeny by selfing, inbreeding or outcrossing, strongly affect important ecological and evolutionary processes. Large variations in mating systems exist in fungi, allowing the study of their origin and consequences. In fungi, sexual incompatibility is determined by molecular recognition mechanisms, controlled by a single mating-type locus in most unifactorial fungi. In Basidiomycete fungi, however, which include rusts, smuts and mushrooms, a system has evolved in which incompatibility is controlled by two unlinked loci. This bifactorial system probably evolved from a unifactorial system. Multiple independent transitions back to a unifactorial system occurred. It is still unclear what force drove evolution and maintenance of these contrasting inheritance patterns that determine mating compatibility. Here, we give an overview of the evolutionary factors that might have driven the evolution of bifactoriality from a unifactorial system and the transitions back to unifactoriality. Bifactoriality most likely evolved for selfing avoidance. Subsequently, multiallelism at mating-type loci evolved through negative frequency-dependent selection by increasing the chance to find a compatible mate. Unifactoriality then evolved back in some species, possibly because either selfing was favoured or for increasing the chance to find a compatible mate in species with few alleles. Owing to the existence of closely related unifactorial and bifactorial species and the increasing knowledge of the genetic systems of the different mechanisms, Basidiomycetes provide an excellent model for studying the different forces that shape breeding systems. PMID:23838688

Modulation and coding for a compatible Discrete Address Beacon System.

DOT National Transportation Integrated Search

1972-02-01

One of several possible candidate configurations for the Discrete Address System is described. The configuration presented is compatible with the Air Traffic Control Radar Beacon System, and it provides for gradual transition from one system to the o...

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.

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

DNA Nanotubes for NMR Structure Determination of Membrane Proteins

PubMed Central

Bellot, Gaëtan; McClintock, Mark A.; Chou, James J; Shih, William M.

2013-01-01

Structure determination of integral membrane proteins by solution NMR represents one of the most important challenges of structural biology. A Residual-Dipolar-Coupling-based refinement approach can be used to solve the structure of membrane proteins up to 40 kDa in size, however, a weak-alignment medium that is detergent-resistant is required. Previously, availability of media suitable for weak alignment of membrane proteins was severely limited. We describe here a protocol for robust, large-scale synthesis of detergent-resistant DNA nanotubes that can be assembled into dilute liquid crystals for application as weak-alignment media in solution NMR structure determination of membrane proteins in detergent micelles. The DNA nanotubes are heterodimers of 400nm-long six-helix bundles each self-assembled from a M13-based p7308 scaffold strand and >170 short oligonucleotide staple strands. Compatibility with proteins bearing considerable positive charge as well as modulation of molecular alignment, towards collection of linearly independent restraints, can be introduced by reducing the negative charge of DNA nanotubes via counter ions and small DNA binding molecules. This detergent-resistant liquid-crystal media offers a number of properties conducive for membrane protein alignment, including high-yield production, thermal stability, buffer compatibility, and structural programmability. Production of sufficient nanotubes for 4–5 NMR experiments can be completed in one week by a single individual. PMID:23518667

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.

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.

Extending hepatocyte functionality for drug-testing applications using high-viscosity alginate-encapsulated three-dimensional cultures in bioreactors.

PubMed

Miranda, Joana P; Rodrigues, Armanda; Tostões, Rui M; Leite, Sofia; Zimmerman, Heiko; Carrondo, Manuel J T; Alves, Paula M

2010-12-01

The maintenance of differentiated hepatocyte phenotype in vitro depends on several factors-in particular, on extracellular matrix interactions, for example, with three-dimensional (3D) matrices. Alginate hydrogel provides the cells with a good extracellular matrix due to the formation of a massive capsule with semi-permeable properties that allows for diffusion of the medium components into the cells as well as efficient waste product elimination. Simultaneously, alginate protects the cells from shear stress caused by the hydrodynamics when cultured in stirred systems such as bioreactors. We have previously developed a hepatocyte aggregate 3D culture system in a bioreactor where improved hepatocyte functionality could be maintained over longer periods (21 days). In this work, ultra-high-viscosity alginate was used for hepatocyte aggregates entrapment. Hepatocyte biotransformation (phase I and II enzymes), CYP450 inducibility, and secretory capacity (albumin and urea production) were monitored. The analyses were performed in both spinner vessels and bioreactors to test the effect of the pO(2) control, unavailable in the spinners. Performance of alginate-encapsulated hepatocyte aggregates in culture was compared with nonencapsulated aggregate cultures in both bioreactor (controlled environment) and spinner vessels. For both culture systems, hepatocytes' metabolic and biotransformation capacities were maintained for up to 1 month, and encapsulated cells in bioreactors showed the best performance. In particular, albumin production rate increased 2- and 1.5-fold in encapsulated aggregates compared with nonencapsulated aggregates in bioreactor and spinner vessels, respectively. Urea production rate increased twofold in encapsulated cultures compared with nonencapsulated cells, in both bioreactor and spinner vessels. Similarly, in both the bioreactor and the spinner system, cell encapsulation resulted in a 1.5- and 2.8-fold improvement of hepatocyte 7-ethoxycoumarin and

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.

Oscillating Cell Culture Bioreactor

NASA Technical Reports Server (NTRS)

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

2010-01-01

dynamic shear (i.e., as required for viability of shear-sensitive cells) to the developing engineered tissue construct. This bioreactor was recently utilized to show independent and interactive effects of a growth factor (IGF-I) and slow bidirectional perfusion on the survival, differentiation, and contractile performance of 3D tissue engineering cardiac constructs. The main application of this system is within the tissue engineering industry. The ideal final application is within the automated mass production of tissue- engineered constructs. Target industries could be both life sciences companies as well as bioreactor device producing companies.

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.

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.

Differentiation of cartilaginous anlage in entire embryonic mouse limbs cultured in a rotating bioreactor.

NASA Astrophysics Data System (ADS)

Duke, P.; Oakley, C.; Montufar-Solis, D.

The embryonic mammalian limb is sensitive both in vivo and in vitro to changes in gravitational force. Hypergravity of centrifugation and microgravity of space decreased size of elements due to precocious or delayed chondrogenesis respectively. In recapitulating spaceflight experiments, premetatarsals were cultured in suspension in a low stress, low sheer rotating bioreactor, and found to be shorter than those cultured in standard culture dishes, and cartilage development was delayed. This study only measured length of the metatarsals, and did not account for possible changes in width and/or in form of the skeletal elements. Shorter cartilage elements in limbbuds cultured in the bioreactor may be due to the ability of the system to reproduce a more in vivo 3D shape than traditional organ cultures. Tissues subjected to traditional organ cultures become flattened by their own weight, attachment to the filter, and restrictions imposed by nutrient diffusion. The purpose of the current experiment was to determine if entire limb buds could be successfully cultured in the bioreactor, and to compare the effects on 3D shape with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were placed either in the bioreactor, in Trowell culture, or fixed as controls. Limbbuds were cultured for six days, fixed, and processed either as whole mounts or embedded for histology. Qualitative analysis revealed that the Trowell culture specimens were flattened, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections of limbbuds from both types of cultures had excellent cartilage differentiation, with apparently more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Morphometric quantitation of the cartilaginous elements for comparisons of the two culture systems was complicated due to some limb buds fusing together during culture. This problem was especially noticeable in the younger limbs, and

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.

A venturi device reduces membrane fouling in a submerged membrane bioreactor.

PubMed

Kayaalp, Necati; Ozturkmen, Gokmen

2016-01-01

In this study, for the first time, a venturi device was integrated into a submerged membrane bioreactor (MBR) to improve membrane surface cleaning and bioreactor oxygenation. The performances of a blower and the venturi device were compared in terms of membrane fouling and bioreactor oxygenation. Upon comparing membrane fouling, the performances were similar for a low operation flux (18 L/m(2).h); however, at a medium flux (32 L/m(2).h), the venturi system operated 3.4 times longer than the blower system, and the final transmembrane pressure was one-third that of the blower system. At the highest flux studied (50 L/m(2).h), the venturi system operated 5.4 times longer than the blower system. The most notable advantage of using a venturi device was that the dissolved oxygen (DO) concentration of the MBR was in the range of 7 to 8 mg/L at a 3 L/min aeration rate, while the DO concentration of the MBR was inadequate (a maximum of 0.29 mg/L) in the blower system. A clean water oxygenation test at a 3 L/min aeration rate indicated that the standard oxygen transfer rate for the venturi system was 9.5 times higher than that of the blower system.

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

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

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

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.

Dissipation of atrazine, enrofloxacin, and sulfamethazine in wood chip bioreactors and impact on denitrification

USDA-ARS?s Scientific Manuscript database

Wood chip bioreactors are receiving increasing attention as a means of reducing nitrate in subsurface tile drainage systems. Agrochemicals in tile drainage water entering wood chip bioreactors can be retained or degraded and may impact denitrification. The degradation of 5 mg L-1 atrazine, enrofloxa...

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.

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.

High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

PubMed Central

Velez‐Suberbie, M. Lourdes; Betts, John P. J.; Walker, Kelly L.; Robinson, Colin; Zoro, Barney

2017-01-01

High throughput automated fermentation systems have become a useful tool in early bioprocess development. In this study, we investigated a 24 x 15 mL single use microbioreactor system, ambr 15f, designed for microbial culture. We compared the fed‐batch growth and production capabilities of this system for two Escherichia coli strains, BL21 (DE3) and MC4100, and two industrially relevant molecules, hGH and scFv. In addition, different carbon sources were tested using bolus, linear or exponential feeding strategies, showing the capacity of the ambr 15f system to handle automated feeding. We used power per unit volume (P/V) as a scale criterion to compare the ambr 15f with 1 L stirred bioreactors which were previously scaled‐up to 20 L with a different biological system, thus showing a potential 1,300 fold scale comparability in terms of both growth and product yield. By exposing the cells grown in the ambr 15f system to a level of shear expected in an industrial centrifuge, we determined that the cells are as robust as those from a bench scale bioreactor. These results provide evidence that the ambr 15f system is an efficient high throughput microbial system that can be used for strain and molecule selection as well as rapid scale‐up. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:58–68, 2018 PMID:28748655

Hollow Fiber Membrane Bioreactor Systems for Wastewater Processing: Effects of Environmental Stresses Including Dormancy Cycling and Antibiotic Dosing

NASA Technical Reports Server (NTRS)

Coutts, Janelle L.; Hummerick, Mary E.; Lunn, Griffin M.; Larson, Brian D.; Spencer, LaShelle E.; Kosiba, Michael L.; Khodadad, Christina L.; Catechis, John A.; Birmele, Michele N.; Wheeler, Raymond M.

2016-01-01

Membrane-aerated biofilm reactors (MABRs) have been studied for a number of years as an alternate approach for treating wastewater streams during space exploration. While the technology provides a promising pre-treatment for lowering organic carbon and nitrogen content without the need for harsh stabilization chemicals, several challenges must be addressed before adoption of the technology in future missions. One challenge is the transportation of bioreactors containing intact, active biofilms as a means for rapid start-up on the International Space Station or beyond. Similarly, there could be a need for placing these biological systems into a dormant state for extended periods when the system is not in use, along with the ability for rapid restart. Previous studies indicated that there was little influence of storage condition (4 or 25 C, with or without bulk fluid) on recovery of bioreactors with immature biofilms (48 days old), but that an extensive recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy within 4 days (approximately 1 residence). Further dormancy and recovery testing is presented here that examines the role of biofilm age on recovery requirements, repeated dormancy cycle capabilities, and effects of long-duration dormancy cycles (8-9 months) on HFMB systems. Another challenge that must be addressed is the possibility of antibiotics entering the wastewater stream. Currently, for most laboratory tests of biological water processors, donors providing urine may not contribute to the study when taking antibiotics because the effects on the system are yet uncharacterized. A simulated urinary tract infection event, where an opportunistic, pathogenic organism, E. coli, was introduced to the HFMBs followed by dosing with an antibiotic, ciprofloxacin, was completed to study the effect of the antibiotic on reactor performance and to also examine the development of

Important operational parameters of membrane bioreactor-sludge disintegration (MBR-SD) system for zero excess sludge production.

PubMed

Yoon, Seong-Hoon

2003-04-01

In order to prevent excess sludge production during wastewater treatment, a membrane bioreactor-sludge disintegration (MBR-SD) system has been introduced, where the disintegrated sludge is recycled to the bioreactor as a feed solution. In this study, a mathematical model was developed by incorporating a sludge disintegration term into the conventional activated sludge model and the relationships among the operational parameters were investigated. A new definition of F/M ratio for the MBR-SD system was suggested to evaluate the actual organic loading rate. The actual F/M ratio was expected to be much higher than the apparent F/M ratio in MBR-SD. The kinetic parameters concerning the biodegradability of organics hardly affect the system performance. Instead, sludge solubilization ratio (alpha) in the SD process and particulate hydrolysis rate constant (k(h)) in biological reaction determine the sludge disintegration number (SDN), which is related with the overall economics of the MBR-SD system. Under reasonable alpha and k(h) values, SDN would range between 3 and 5 which means the amount of sludge required to be disintegrated would be 3-5 times higher for preventing a particular amount of sludge production. Finally, normalized sludge disintegration rate (q/V) which is needed to maintain a certain level of MLSS in the MBR-SD system was calculated as a function of F/V ratio.

Thermal compatibility of dental ceramic systems using cylindrical and spherical geometries.

PubMed

DeHoff, Paul H; Barrett, Allyson A; Lee, Robert B; Anusavice, Kenneth J

2008-06-01

To test the hypothesis that bilayer ceramic cylinders and spheres can provide valid confirmation of thermal incompatibility stresses predicted by finite element analyses. A commercial core ceramic and an experimental core ceramic were used to fabricate open-ended cylinders and core ceramic spheres. The core cylinders and spheres were veneered with one of four commercial dental ceramics representing four thermally compatible groups and four thermally incompatible groups. Axisymmetric thermal and viscoelastic elements in the ANSYS finite element program were used to calculate temperatures and stresses for each geometry and ceramic combination. This process required a transient heat transfer analysis for each combination to determine input temperatures for the structural model. After fabrication, each specimen was examined visually using fiberoptic transillumination for evidence of cracking. There were 100% failures of the thermally incompatible cylinders while none of the thermally compatible combinations failed. Among the spheres, 100% of the thermally incompatible systems failed, 16% of one of the thermally compatible systems failed, and none of the remaining compatible combinations failed. The calculated stress values were in general agreement with the experimental observations, i.e., low residual stresses for the specimens that did not fail and high residual stresses for the specimens that did fail. Simple screening geometries can be used to identify highly incompatible ceramic combinations, but they do not identify marginally incompatible systems.

Thermal compatibility of dental ceramic systems using cylindrical and spherical geometries

PubMed Central

DeHoff, Paul H.; Barrett, Allyson A.; Lee, Robert B.; Anusavice, Kenneth J.

2009-01-01

Objective To test the hypothesis that bilayer ceramic cylinders and spheres can provide valid confirmation of thermal incompatibility stresses predicted by finite element analyses. Methods A commercial core ceramic and an experimental core ceramic were used to fabricate open-ended cylinders and core ceramic spheres. The core cylinders and spheres were veneered with one of four commercial dental ceramics representing four thermally compatible groups and four thermally incompatible groups. Axisymmetric thermal and viscoelastic elements in the ANSYS finite element program were used to calculate temperatures and stresses for each geometry and ceramic combination. This process required a transient heat transfer analysis for each combination to determine input temperatures for the structural model. Results After fabrication, each specimen was examined visually using fiberoptic transillumination for evidence of cracking. There were 100% failures of the thermally incompatible cylinders while none of the thermally compatible combinations failed. Among the spheres, 100% of the thermally incompatible systems failed, 16% of one of the thermally compatible systems failed, and none of the remaining compatible combinations failed. The calculated stress values were in general agreement with the experimental observations, i.e., low residual stresses for the specimens that did not fail and high residual stresses for the specimens that did fail. Significance Simple screening geometries can be used to identify highly incompatible ceramic combinations, but they do not identify marginally incompatible systems. PMID:17949805

EMC (Electromagnetic Compatibility) System Test and Analysis Interface.

DTIC Science & Technology

1983-05-01

D- R136 64 EMC (ELECTROARGNETIC COMPATIBILITY) SYSTEM TEST AND V ANALYSIS INTE FACE(J) BOEING CO SEATTLE WA E F BALL ET AL MAY 93 RRDC-TR-83-121...RADC-TR-83- 121 Final Tedical Report May 1963 EMC SYSTEM TEST AND ANAL YSIS INTERFACE_ The Boeing Company E. F. Ball, L. Knutson and B. L...Carlson *. . APPROVE FOR PUBLIC REESEk DIS TRIBUTION ULMTED D IS -ELECTE ,... DEC 20 1983 >- D" c: ROME AIR DEVELOPMENT CENTER * Air Force Systems Command

Quantitative analysis of Earth's field NMR spectra of strongly-coupled heteronuclear systems.

PubMed

Halse, Meghan E; Callaghan, Paul T; Feland, Brett C; Wasylishen, Roderick E

2009-09-01

In the Earth's magnetic field, it is possible to observe spin systems consisting of unlike spins that exhibit strongly coupled second-order NMR spectra. Such spectra result when the J-coupling between two unlike spins is of the same order of magnitude as the difference in their Larmor precession frequencies. Although the analysis of second-order spectra involving only spin-(1/2) nuclei has been discussed since the early days of NMR spectroscopy, NMR spectra involving spin-(1/2) nuclei and quadrupolar (I>(1/2)) nuclei have rarely been treated. Two examples are presented here, the tetrahydroborate anion, BH4-, and the ammonium cation, NH4+. For the tetrahydroborate anion, (1)J((11)B,(1)H)=80.9Hz, and in an Earth's field of 53.3microT, nu((1)H)=2269Hz and nu((11)B)=728Hz. The (1)H NMR spectra exhibit features that both first- and second-order perturbation theory are unable to reproduce. On the other hand, second-order perturbation theory adequately describes (1)H NMR spectra of the ammonium anion, (14)NH4+, where (1)J((14)N,(1)H)=52.75Hz when nu((1)H)=2269Hz and nu((14)N)=164Hz. Contrary to an early report, we find that the (1)H NMR spectra are independent of the sign of (1)J((14)N,(1)H). Exact analysis of two-spin systems consisting of quadrupolar nuclei and spin-(1/2) nuclei are also discussed.

Nomenclature associated with chemical characterization of and compatibility evaluations for medical product delivery systems.

PubMed

Jenke, Dennis R

2003-01-01

Delivery systems are used to store, contain, and/or administer liquid pharmaceutical products. Gaining an understanding of the chemical composition of such a delivery system is necessary with respect to effective system development, registration, and production. Additionally, the ability of the delivery system to impact the chemical composition of the contacted product may define the safety and/or efficacy of the product. Assessing the compatibility of the delivery system and the product is thus both necessary and desirable. The nomenclature associated with compatibility assessments has not been standardized, oftimes leading to conflicting or confusing information. This manuscript puts forth a nomenclature which classifies those chemical entities which participate in the system/product interaction and delineates the various extraction strategies which may be used in compatibility assessments.

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.

Manufacturing recombinant proteins in kg-ton quantities using animal cells in bioreactors.

PubMed

De Jesus, Maria; Wurm, Florian M

2011-06-01

Mammalian cells in bioreactors as production host are the focus of this review. We wish to briefly describe today's technical status and to highlight emerging trends in the manufacture of recombinant therapeutic proteins, focusing on Chinese hamster ovary (CHO) cells. CHO cells are the manufacturing host system of choice for more than 70% of protein pharmaceuticals on the market [21]. The current global capacity to grow mammalian cells in bioreactors stands at about 0.5 million liters, whereby the largest vessels can have a working volume of about 20,000l. We are focusing in this article on the upstream part of protein manufacturing. Over the past 25 years, volumetric yields for recombinant cell lines have increased about 20-fold mainly as the result of improvements in media and bioprocess design. Future yield increases are expected to come from improved gene delivery methods, from improved, possibly genetically modified host systems, and from further improved bioprocesses in bioreactors. Other emerging trends in protein manufacturing that are discussed include the use of disposal bioreactors and transient gene expression. We specifically highlight here current research in our own laboratories. Copyright © 2011 Elsevier B.V. All rights reserved.

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.

A perfusion bioreactor system efficiently generates cell-loaded bone substitute materials for addressing critical size bone defects.

PubMed

Kleinhans, Claudia; Mohan, Ramkumar Ramani; Vacun, Gabriele; Schwarz, Thomas; Haller, Barbara; Sun, Yang; Kahlig, Alexander; Kluger, Petra; Finne-Wistrand, Anna; Walles, Heike; Hansmann, Jan

2015-09-01

Critical size bone defects and non-union fractions are still challenging to treat. Cell-loaded bone substitutes have shown improved bone ingrowth and bone formation. However, a lack of methods for homogenously colonizing scaffolds limits the maximum volume of bone grafts. Additionally, therapy robustness is impaired by heterogeneous cell populations after graft generation. Our aim was to establish a technology for generating grafts with a size of 10.5 mm in diameter and 25 mm of height, and thus for grafts suited for treatment of critical size bone defects. Therefore, a novel tailor-made bioreactor system was developed, allowing standardized flow conditions in a porous poly(L-lactide-co-caprolactone) material. Scaffolds were seeded with primary human mesenchymal stem cells derived from four different donors. In contrast to static experimental conditions, homogenous cell distributions were accomplished under dynamic culture. Additionally, culture in the bioreactor system allowed the induction of osteogenic lineage commitment after one week of culture without addition of soluble factors. This was demonstrated by quantitative analysis of calcification and gene expression markers related to osteogenic lineage. In conclusion, the novel bioreactor technology allows efficient and standardized conditions for generating bone substitutes that are suitable for the treatment of critical size defects in humans. © 2015 The Authors. Biotechnology Journal published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial Licence, which permits use, distribution and reproduction in any medium, provided the Contribution is properly cited and is not used for commercial purpose.

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.

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.

Successful chondrogenesis within scaffolds, using magnetic stem cell confinement and bioreactor maturation.

PubMed

Luciani, Nathalie; Du, Vicard; Gazeau, Florence; Richert, Alain; Letourneur, Didier; Le Visage, Catherine; Wilhelm, Claire

2016-06-01

Tissue engineering strategies, such as cellularized scaffolds approaches, have been explored for cartilage replacement. The challenge, however, remains to produce a cartilaginous tissue incorporating functional chondrocytes and being large and thick enough to be compatible with the replacement of articular defects. Here, we achieved unprecedented cartilage tissue production into a porous polysaccharide scaffold by combining of efficient magnetic condensation of mesenchymal stem cells, and dynamic maturation in a bioreactor. In optimal conditions, all the hallmarks of chondrogenesis were enhanced with a 50-fold increase in collagen II expression compared to negative control, an overexpression of aggrecan and collagen XI, and a very low expression of collagen I and RUNX2. Histological staining showed a large number of cellular aggregates, as well as an increased proteoglycan synthesis by chondrocytes. Interestingly, electron microscopy showed larger chondrocytes and a more abundant extracellular matrix. In addition, the periodicity of the neosynthesized collagen fibers matched that of collagen II. These results represent a major step forward in replacement tissue for cartilage defects. A combination of several innovative technologies (magnetic cell seeding, polysaccharide porous scaffolds, and dynamic maturation in bioreactor) enabled unprecedented successful chondrogenesis within scaffolds. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

DNA nanotubes for NMR structure determination of membrane proteins.

PubMed

Bellot, Gaëtan; McClintock, Mark A; Chou, James J; Shih, William M

2013-04-01

Finding a way to determine the structures of integral membrane proteins using solution nuclear magnetic resonance (NMR) spectroscopy has proved to be challenging. A residual-dipolar-coupling-based refinement approach can be used to resolve the structure of membrane proteins up to 40 kDa in size, but to do this you need a weak-alignment medium that is detergent-resistant and it has thus far been difficult to obtain such a medium suitable for weak alignment of membrane proteins. We describe here a protocol for robust, large-scale synthesis of detergent-resistant DNA nanotubes that can be assembled into dilute liquid crystals for application as weak-alignment media in solution NMR structure determination of membrane proteins in detergent micelles. The DNA nanotubes are heterodimers of 400-nm-long six-helix bundles, each self-assembled from a M13-based p7308 scaffold strand and >170 short oligonucleotide staple strands. Compatibility with proteins bearing considerable positive charge as well as modulation of molecular alignment, toward collection of linearly independent restraints, can be introduced by reducing the negative charge of DNA nanotubes using counter ions and small DNA-binding molecules. This detergent-resistant liquid-crystal medium offers a number of properties conducive for membrane protein alignment, including high-yield production, thermal stability, buffer compatibility and structural programmability. Production of sufficient nanotubes for four or five NMR experiments can be completed in 1 week by a single individual.

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.).

76 FR 46798 - Compatibility of Underground Storage Tank Systems With Biofuel Blends; Correction

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-03

... ENVIRONMENTAL PROTECTION AGENCY [EPA-HQ-UST-2010-0651; FRL-9447-3] Compatibility of Underground Storage Tank Systems With Biofuel Blends; Correction AGENCY: Environmental Protection Agency (EPA). ACTION... of underground storage tanks (USTs) can demonstrate compliance with the Federal compatibility...

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...

Statistical EMC: A new dimension electromagnetic compatibility of digital electronic systems

NASA Astrophysics Data System (ADS)

Tsaliovich, Anatoly

Electromagnetic compatibility compliance test results are used as a database for addressing three classes of electromagnetic-compatibility (EMC) related problems: statistical EMC profiles of digital electronic systems, the effect of equipment-under-test (EUT) parameters on the electromagnetic emission characteristics, and EMC measurement specifics. Open area test site (OATS) and absorber line shielded room (AR) results are compared for equipment-under-test highest radiated emissions. The suggested statistical evaluation methodology can be utilized to correlate the results of different EMC test techniques, characterize the EMC performance of electronic systems and components, and develop recommendations for electronic product optimal EMC design.

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

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.

New optical architecture for holographic data storage system compatible with Blu-ray Disc™ system

NASA Astrophysics Data System (ADS)

Shimada, Ken-ichi; Ide, Tatsuro; Shimano, Takeshi; Anderson, Ken; Curtis, Kevin

2014-02-01

A new optical architecture for holographic data storage system which is compatible with a Blu-ray Disc™ (BD) system is proposed. In the architecture, both signal and reference beams pass through a single objective lens with numerical aperture (NA) 0.85 for realizing angularly multiplexed recording. The geometry of the architecture brings a high affinity with an optical architecture in the BD system because the objective lens can be placed parallel to a holographic medium. Through the comparison of experimental results with theory, the validity of the optical architecture was verified and demonstrated that the conventional objective lens motion technique in the BD system is available for angularly multiplexed recording. The test-bed composed of a blue laser system and an objective lens of the NA 0.85 was designed. The feasibility of its compatibility with BD is examined through the designed test-bed.

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.

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.

Cooperation, Convertibility, and Compatibility Among Information Systems: A Literature Review.

ERIC Educational Resources Information Center

Henderson, Madeline M.; And Others

The purpose of the study of the literature on which this report is based was to examine those problems in the field of documentation and in the operation of information systems which could possibly be solved or alleviated by some greater measure of cooperation, convertibility, or compatibility among systems, particularly those systems for handling…

High throughput automated microbial bioreactor system used for clone selection and rapid scale-down process optimization.

PubMed

Velez-Suberbie, M Lourdes; Betts, John P J; Walker, Kelly L; Robinson, Colin; Zoro, Barney; Keshavarz-Moore, Eli

2018-01-01

High throughput automated fermentation systems have become a useful tool in early bioprocess development. In this study, we investigated a 24 x 15 mL single use microbioreactor system, ambr 15f, designed for microbial culture. We compared the fed-batch growth and production capabilities of this system for two Escherichia coli strains, BL21 (DE3) and MC4100, and two industrially relevant molecules, hGH and scFv. In addition, different carbon sources were tested using bolus, linear or exponential feeding strategies, showing the capacity of the ambr 15f system to handle automated feeding. We used power per unit volume (P/V) as a scale criterion to compare the ambr 15f with 1 L stirred bioreactors which were previously scaled-up to 20 L with a different biological system, thus showing a potential 1,300 fold scale comparability in terms of both growth and product yield. By exposing the cells grown in the ambr 15f system to a level of shear expected in an industrial centrifuge, we determined that the cells are as robust as those from a bench scale bioreactor. These results provide evidence that the ambr 15f system is an efficient high throughput microbial system that can be used for strain and molecule selection as well as rapid scale-up. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:58-68, 2018. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

Fiber Attachment Module Experiment (FAME): Using a Multiplexed Miniature Hollow Fiber Membrane Bioreactor Solution for Rapid Process Testing

NASA Technical Reports Server (NTRS)

Coutts, Janelle L.; Lunn, Griffin M.; Koss, Lawrence L.; Hummerick, Mary E.; Spencer, Lachelle E.; Johnsey, Marissa N.; Richards, Jeffrey T.; Ellis, Ronald; Birmele, Michele N.; Wheeler, Raymond M.

2014-01-01

Bioreactor research is mostly limited to continuous stirred-tank reactors (CSTRs) which are not an option for microgravity (g) applications due to the lack of a gravity gradient to drive aeration as described by the Archimedes principle. Bioreactors and filtration systems for treating wastewater in g could avoid the need for harsh pretreatment chemicals and improve overall water recovery. Solution: Membrane Aerated Bioreactors (MABRs) for g applications, including possible use for wastewater treatment systems for the International Space Station (ISS).

X-ray Phase Contrast Imaging of Calcified Tissue and Biomaterial Structure in Bioreactor Engineered Tissues

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

Appel, Alyssa A.; Larson, Jeffery C.; Garson, III, Alfred B.

2014-11-04

Tissues engineered in bioreactor systems have been used clinically to replace damaged tissues and organs. In addition, these systems are under continued development for many tissue engineering applications. The ability to quantitatively assess material structure and tissue formation is critical for evaluating bioreactor efficacy and for preimplantation assessment of tissue quality. These techniques allow for the nondestructive and longitudinal monitoring of large engineered tissues within the bioreactor systems and will be essential for the translation of these strategies to viable clinical therapies. X-ray Phase Contrast (XPC) imaging techniques have shown tremendous promise for a number of biomedical applications owing tomore » their ability to provide image contrast based on multiple X-ray properties, including absorption, refraction, and scatter. In this research, mesenchymal stem cell-seeded alginate hydrogels were prepared and cultured under osteogenic conditions in a perfusion bioreactor. The constructs were imaged at various time points using XPC microcomputed tomography (µCT). Imaging was performed with systems using both synchrotron- and tube-based X-ray sources. XPC µCT allowed for simultaneous three-dimensional (3D) quantification of hydrogel size and mineralization, as well as spatial information on hydrogel structure and mineralization. Samples were processed for histological evaluation and XPC showed similar features to histology and quantitative analysis consistent with the histomorphometry. Furthermore, these results provide evidence of the significant potential of techniques based on XPC for noninvasive 3D imaging engineered tissues grown in bioreactors.« less

A miniaturized bioreactor system for the evaluation of cell interaction with designed substrates in perfusion culture.

PubMed

Sun, T; Donoghue, P S; Higginson, J R; Gadegaard, N; Barnett, S C; Riehle, M O

2012-12-01

In tissue engineering, chemical and topographical cues are normally developed using static cell cultures but then applied directly to tissue cultures in three dimensions (3D) and under perfusion. As human cells are very sensitive to changes in the culture environment, it is essential to evaluate the performance of any such cues in a perfused environment before they are applied to tissue engineering. Thus, the aim of this research was to bridge the gap between static and perfusion cultures by addressing the effect of perfusion on cell cultures within 3D scaffolds. For this we developed a scaled-down bioreactor system, which allows evaluation of the effectiveness of various chemical and topographical cues incorporated into our previously developed tubular ε-polycaprolactone scaffold under perfused conditions. Investigation of two exemplary cell types (fibroblasts and cortical astrocytes) using the miniaturized bioreactor indicated that: (a) quick and firm cell adhesion in the 3D scaffold was critical for cell survival in perfusion culture compared with static culture; thus, cell-seeding procedures for static cultures might not be applicable, therefore it was necessary to re-evaluate cell attachment on different surfaces under perfused conditions before a 3D scaffold was applied for tissue cultures; (b) continuous medium perfusion adversely influenced cell spread and survival, which could be balanced by intermittent perfusion; (c) micro-grooves still maintained their influences on cell alignment under perfused conditions, while medium perfusion demonstrated additional influence on fibroblast alignment but not on astrocyte alignment on grooved substrates. This research demonstrated that the mini-bioreactor system is crucial for the development of functional scaffolds with suitable chemical and topographical cues by bridging the gap between static culture and perfusion culture. Copyright © 2011 John Wiley & Sons, Ltd.

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

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.

Design and Use of a Novel Bioreactor for Regeneration of Biaxially Stretched Tissue-Engineered Vessels

PubMed Central

Huang, Angela Hai; Lee, Yong-Ung; Calle, Elizabeth A.; Boyle, Michael; Starcher, Barry C.; Humphrey, Jay D.

2015-01-01

Conventional bioreactors are used to enhance extracellular matrix (ECM) production and mechanical strength of tissue-engineered vessels (TEVs) by applying circumferential strain, which is uniaxial stretching. However, the resulting TEVs still suffer from inadequate mechanical properties, where rupture strengths and compliance values are still very different from native arteries. The biomechanical milieu of native arteries consists of both circumferential and axial loading. Therefore, to better simulate the physiological stresses acting on native arteries, we built a novel bioreactor system to enable biaxial stretching of engineered arteries during culture. This new bioreactor system allows for independent control of circumferential and axial stretching parameters, such as displacement and beat rate. The assembly and setup processes for this biaxial bioreactor system are reliable with a success rate greater than 75% for completion of long-term sterile culture. This bioreactor also supports side-by-side assessments of TEVs that are cultured under three types of mechanical conditions (static, uniaxial, and biaxial), all within the same biochemical environment. Using this bioreactor, we examined the impact of biaxial stretching on arterial wall remodeling of TEVs. Biaxial TEVs developed the greatest wall thickness compared with static and uniaxial TEVs. Unlike uniaxial loading, biaxial loading led to undulated collagen fibers that are commonly found in native arteries. More importantly, the biaxial TEVs developed the most mature elastin in the ECM, both qualitatively and quantitatively. The presence of mature extracellular elastin along with the undulated collagen fibers may contribute to the observed vascular compliance in the biaxial TEVs. The current work shows that biaxial stretching is a novel and promising means to improve TEV generation. Furthermore, this novel system allows us to optimize biomechanical conditioning by unraveling the interrelationships among the

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.

Visualizing feasible operating ranges within tissue engineering systems using a "windows of operation" approach: a perfusion-scaffold bioreactor case study.

PubMed

McCoy, Ryan J; O'Brien, Fergal J

2012-12-01

Tissue engineering approaches to developing functional substitutes are often highly complex, multivariate systems where many aspects of the biomaterials, bio-regulatory factors or cell sources may be controlled in an effort to enhance tissue formation. Furthermore, success is based on multiple performance criteria reflecting both the quantity and quality of the tissue produced. Managing the trade-offs between different performance criteria is a challenge. A "windows of operation" tool that graphically represents feasible operating spaces to achieve user-defined levels of performance has previously been described by researchers in the bio-processing industry. This paper demonstrates the value of "windows of operation" to the tissue engineering field using a perfusion-scaffold bioreactor system as a case study. In our laboratory, perfusion bioreactor systems are utilized in the context of bone tissue engineering to enhance the osteogenic differentiation of cell-seeded scaffolds. A key challenge of such perfusion bioreactor systems is to maximize the induction of osteogenesis but minimize cell detachment from the scaffold. Two key operating variables that influence these performance criteria are the mean scaffold pore size and flow-rate. Using cyclooxygenase-2 and osteopontin gene expression levels as surrogate indicators of osteogenesis, we employed the "windows of operation" methodology to rapidly identify feasible operating ranges for the mean scaffold pore size and flow-rate that achieved user-defined levels of performance for cell detachment and differentiation. Incorporation of such tools into the tissue engineer's armory will hopefully yield a greater understanding of the highly complex systems used and help aid decision making in future translation of products from the bench top to the market place. Copyright © 2012 Wiley Periodicals, Inc.

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.

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

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.

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

Compatibility of Niobium Alloys and Superalloys in a Flowing He-Xe Power Conversion System

NASA Technical Reports Server (NTRS)

Bowman, Cheryl L.; Ritzert, Frank J.; Smialek, James L.; Jaster, Mark L.; rker, Samuel P.

2004-01-01

Proposed concepts for an ambitious mission to explore Jupiter's three icy moons place significant demands on the various spacecraft systems. There are many challenges related to the high output power conversion systems being considered, and one example is the need to ensure system compatibility at all levels. The utilization of appropriate materials for component structures is important to ensuring long mission life. Refractory metal alloys have attractive high-temperature properties in inert environments, but these alloys are sometimes susceptible to contamination. Potential material compatibility issues exist between refractory metal candidates and more conventional alloys. Nb-1Zr has long been considered one of the most well characterized refractory alloys that is well suited for elevated-temperature use and liquid-metal compatibility. However, previous studies have suggested that niobium alloys can not co-exist in a closed system with traditional stainless steels or superalloys due to transport of contaminants. The relevance of this information to a proposed power conversion system is discussed. Also, experiments and fundamental calculations are being performed to determine contamination transport from candidate superalloys to Nb-1Zr in a closed system with an inert carrier gas. Potential protective schemes are explored to ensure system level compatibility between the refractory alloy Nb-1Zr and a nickel-based superalloy.

Improved function and growth of pancreatic cells in a three-dimensional bioreactor environment.

PubMed

Samuelson, Lisa; Gerber, David A

2013-01-01

Methods of three-dimensional (3D) cell culture have made significant progress in recent years due to a better understanding of cell to cell interactions and the cell's interface with their surrounding environment. We hypothesized that a microgravity 3D culture system would improve upon the growth and function of a pancreatic progenitor cell population. We developed a rotating wall vessel bioreactor and established a culture system using a pancreatic cell line. Cells in the bioreactors showed robust proliferation, enhanced transcriptional signaling, and improved translation of pancreatic genes compared with two-dimensional static culture. Cells also gained the ability to respond to glucose stimulation, which was not observed in the control cultures. These findings suggest that a 3D microgravity bioreactor environment mimics the niche of the pancreas yielding a cell source with potential for cell-based therapy in the treatment of diabetes.

Improvement of In Vitro Three‐Dimensional Cartilage Regeneration by a Novel Hydrostatic Pressure Bioreactor

PubMed Central

Chen, Jie; Yuan, Zhaoyuan; Liu, Yu; Zheng, Rui; Dai, Yao; Tao, Ran; Xia, Huitang; Liu, Hairong; Zhang, Zhiyong; Zhang, Wenjie; Liu, Wei; Cao, Yilin

2016-01-01

Abstract In vitro three‐dimensional (3D) cartilage regeneration is a promising strategy for repair of cartilage defects. However, inferior mechanical strength and tissue homogeneity greatly restricted its clinical translation. Simulation of mechanical stress through a bioreactor is an important approach for improving in vitro cartilage regeneration. The current study developed a hydrostatic pressure (HP) bioreactor based on a novel pressure‐transmitting mode achieved by slight deformation of a flexible membrane in a completely sealed stainless steel device. The newly developed bioreactor efficiently avoided the potential risks of previously reported pressure‐transmitting modes and simultaneously addressed a series of important issues, such as pressure scopes, culture chamber sizes, sealability, contamination control, and CO2 balance. The whole bioreactor system realized stable long‐term (8 weeks) culture under high HP (5–10 MPa) without the problems of medium leakage and contamination. Furthermore, the results of in vitro 3D tissue culture based on a cartilage regeneration model revealed that HP provided by the newly developed bioreactor efficiently promoted in vitro 3D cartilage formation by improving its mechanical strength, thickness, and homogeneity. Detailed analysis in cell proliferation, cartilage matrix production, and cross‐linking level of collagen macromolecules, as well as density and alignment of collagen fibers, further revealed the possible mechanisms that HP regulated in vitro cartilage regeneration. The current study provided a highly efficient and stable bioreactor system for improving in vitro 3D cartilage regeneration and thus will help to accelerate its clinical translation. Stem Cells Translational Medicine 2017;6:982–991 PMID:28297584

Process for whole cell saccharification of lignocelluloses to sugars using a dual bioreactor system

DOEpatents

Lu, Jue [Okemos, MI; Okeke, Benedict [Montgomery, AL

2012-03-27

The present invention describes a process for saccharification of lignocelluloses to sugars using whole microbial cells, which are enriched from cultures inoculated with paper mill waste water, wood processing waste and soil. A three-member bacterial consortium is selected as a potent microbial inocula and immobilized on inedible plant fibers for biomass saccharification. The present invention further relates the design of a dual bioreactor system, with various biocarriers for enzyme immobilization and repeated use. Sugars are continuously removed eliminating end-product inhibition and consumption by cell.

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.

It's all in the timing: modeling isovolumic contraction through development and disease with a dynamic dual electromechanical bioreactor system.

PubMed

Morgan, Kathy Ye; Black, Lauren Deems

2014-01-01

This commentary discusses the rationale behind our recently reported work entitled "Mimicking isovolumic contraction with combined electromechanical stimulation improves the development of engineered cardiac constructs," introduces new data supporting our hypothesis, and discusses future applications of our bioreactor system. The ability to stimulate engineered cardiac tissue in a bioreactor system that combines both electrical and mechanical stimulation offers a unique opportunity to simulate the appropriate dynamics between stretch and contraction and model isovolumic contraction in vitro. Our previous study demonstrated that combined electromechanical stimulation that simulated the timing of isovolumic contraction in healthy tissue improved force generation via increased contractile and calcium handling protein expression and improved hypertrophic pathway activation. In new data presented here, we further demonstrate that modification of the timing between electrical and mechanical stimulation to mimic a non-physiological process negatively impacts the functionality of the engineered constructs. We close by exploring the various disease states that have altered timing between the electrical and mechanical stimulation signals as potential future directions for the use of this 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.

Impact of media and antifoam selection on monoclonal antibody production and quality using a high throughput micro‐bioreactor system

PubMed Central

Velugula‐Yellela, Sai Rashmika; Williams, Abasha; Trunfio, Nicholas; Hsu, Chih‐Jung; Chavez, Brittany; Yoon, Seongkyu

2017-01-01

Monoclonal antibody production in commercial scale cell culture bioprocessing requires a thorough understanding of the engineering process and components used throughout manufacturing. It is important to identify high impact components early on during the lifecycle of a biotechnology‐derived product. While cell culture media selection is of obvious importance to the health and productivity of mammalian bioreactor operations, other components such as antifoam selection can also play an important role in bioreactor cell culture. Silicone polymer‐based antifoams were known to have negative impacts on cell health, production, and downstream filtration and purification operations. High throughput screening in micro‐scale bioreactors provides an efficient strategy to identify initial operating parameters. Here, we utilized a micro‐scale parallel bioreactor system to study an IgG1 producing CHO cell line, to screen Dynamis, ProCHO5, PowerCHO2, EX‐Cell Advanced, and OptiCHO media, and 204, C, EX‐Cell, SE‐15, and Y‐30 antifoams and their impacts on IgG1 production, cell growth, aggregation, and process control. This study found ProCHO5, EX‐Cell Advanced, and PowerCHO2 media supported strong cellular growth profiles, with an IVCD of 25‐35 × 106 cells‐d/mL, while maintaining specific antibody production (Qp > 2 pg/cell‐d) for our model cell line and a monomer percentage above 94%. Antifoams C, EX‐Cell, and SE‐15 were capable of providing adequate control of foaming while antifoam 204 and Y‐30 noticeably stunted cellular growth. This work highlights the utility of high throughput micro bioreactors and the importance of identifying both positive and negative impacts of media and antifoam selection on a model IgG1 producing CHO cell line. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:262–270, 2018 PMID:29086492

A nanoliter volume nuclear magnetic resonance (NMR) system using tunneling magneto-resistive (TMR) sensors to recognize biomolecules

NASA Astrophysics Data System (ADS)

Gomez, Pablo

The need to incorporate advanced engineering tools in biology, biochemistry and medicine is in great demand. Many of the existing instruments and tools are usually expensive and require special facilities. With the advent of nanotechnology in the past decade, new approaches to develop devices and tools have been generated by academia and industry. One such technology, NMR spectroscopy, has been used by biochemists for more than 2 decades to study the molecular structure of chemical compounds. However, NMR spectrometers are very expensive and require special laboratory rooms for their proper operation. High magnetic fields with strengths in the order of several Tesla make these instruments unaffordable to most research groups. This doctoral research proposes a new technology to develop NMR spectrometers that can operate at field strengths of less than 0.5 Tesla using an inexpensive permanent magnet and spin dependent nanoscale magnetic devices. This portable NMR system is intended to analyze samples as small as a few nanoliters. The main problem to resolve when downscaling the variables is to obtain an NMR signal with high Signal-To-Noise-Ratio (SNR). A special Tunneling Magneto-Resistive (TMR) sensor design was developed to achieve this goal. The minimum specifications for each component of the proposed NMR system were established. A complete NMR system was designed based on these minimum requirements. The goat was always to find cost effective realistic components. The novel design of the NMR system uses technologies such as Direct Digital Synthesis (DDS), Digital Signal Processing (DSP) and a special Backpropagation Neural Network that finds the best match of the NMR spectrum. The system was designed, calculated and simulated with excellent results. In addition, a general method to design TMR Sensors was developed. The technique was automated and a computer program was written to help the designer perform this task interactively.

Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor

NASA Astrophysics Data System (ADS)

Montufar-Solis, D.; Oakley, C. R.; Jefferson, Y.; Duke, P. J.

2003-10-01

Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (lg). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of El 1

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.

Culturing and applications of rotating wall vessel bioreactor derived 3D epithelial cell models.

PubMed

Radtke, Andrea L; Herbst-Kralovetz, Melissa M

2012-04-03

Cells and tissues in the body experience environmental conditions that influence their architecture, intercellular communications, and overall functions. For in vitro cell culture models to accurately mimic the tissue of interest, the growth environment of the culture is a critical aspect to consider. Commonly used conventional cell culture systems propagate epithelial cells on flat two-dimensional (2-D) impermeable surfaces. Although much has been learned from conventional cell culture systems, many findings are not reproducible in human clinical trials or tissue explants, potentially as a result of the lack of a physiologically relevant microenvironment. Here, we describe a culture system that overcomes many of the culture condition boundaries of 2-D cell cultures, by using the innovative rotating wall vessel (RWV) bioreactor technology. We and others have shown that organotypic RWV-derived models can recapitulate structure, function, and authentic human responses to external stimuli similarly to human explant tissues (1-6). The RWV bioreactor is a suspension culture system that allows for the growth of epithelial cells under low physiological fluid shear conditions. The bioreactors come in two different formats, a high-aspect rotating vessel (HARV) or a slow-turning lateral vessel (STLV), in which they differ by their aeration source. Epithelial cells are added to the bioreactor of choice in combination with porous, collagen-coated microcarrier beads (Figure 1A). The cells utilize the beads as a growth scaffold during the constant free fall in the bioreactor (Figure 1B). The microenvironment provided by the bioreactor allows the cells to form three-dimensional (3-D) aggregates displaying in vivo-like characteristics often not observed under standard 2-D culture conditions (Figure 1D). These characteristics include tight junctions, mucus production, apical/basal orientation, in vivo protein localization, and additional epithelial cell-type specific properties

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. ...

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.

A clinic compatible, open source electrophysiology system.

PubMed

Hermiz, John; Rogers, Nick; Kaestner, Erik; Ganji, Mehran; Cleary, Dan; Snider, Joseph; Barba, David; Dayeh, Shadi; Halgren, Eric; Gilja, Vikash

2016-08-01

Open source electrophysiology (ephys) recording systems have several advantages over commercial systems such as customization and affordability enabling more researchers to conduct ephys experiments. Notable open source ephys systems include Open-Ephys, NeuroRighter and more recently Willow, all of which have high channel count (64+), scalability, and advanced software to develop on top of. However, little work has been done to build an open source ephys system that is clinic compatible, particularly in the operating room where acute human electrocorticography (ECoG) research is performed. We developed an affordable (<; $10,000) and open system for research purposes that features power isolation for patient safety, compact and water resistant enclosures and 256 recording channels sampled up to 20ksam/sec, 16-bit. The system was validated by recording ECoG with a high density, thin film device for an acute, awake craniotomy study at UC San Diego, Thornton Hospital Operating Room.

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.

Spiral vane bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R. (Inventor)

1991-01-01

A spiral vane bioreactor of a perfusion type is described in which a vertical chamber, intended for use in a microgravity condition, has a central rotating filter assembly and has flexible membranes disposed to rotate annularly about the filter assembly. The flexible members have end portions disposed angularly with respect to one another. A fluid replenishment medium is input from a closed loop liquid system to a completely liquid filled chamber containing microcarrier beads, cells and a fluid medium. Output of spent medium is to the closed loop. In the closed loop, the output and input parameters are sensed by sensors. A manifold permits recharging of the nutrients and pH adjustment. Oxygen is supplied and carbon dioxide and bubbles are removed and the system is monitored and controlled by a microprocessor.

Design and testing of a unique randomized gravity, continuous flow bioreactor

NASA Technical Reports Server (NTRS)

Lassiter, Carroll B.

1993-01-01

A rotating, null gravity simulator, or Couette bioreactor was successfully used for the culture of mammalian cells in a simulated microgravity environment. Two limited studies using Lipomyces starkeyi and Streptomyces clavuligerus were also conducted under conditions of simulated weightlessness. Although these studies with microorganisms showed promising preliminary results, oxygen limitations presented significant limitations in studying the biochemical and cultural characteristics of these cell types. Microbial cell systems such as bacteria and yeast promise significant potential as investigative models to study the effects of microgravity on membrane transport, as well as substrate induction of inactive enzyme systems. Additionally, the smaller size of the microorganisms should further reduce the gravity induced oscillatory particle motion and thereby improve the microgravity simulation on earth. Focus is on the unique conceptual design, and subsequent development of a rotating bioreactor that is compatible with the culture and investigation of microgravity effects on microbial systems. The new reactor design will allow testing of highly aerobic cell types under simulated microgravity conditions. The described reactor affords a mechanism for investigating the long term effects of reduced gravity on cellular respiration, membrane transfer, ion exchange, and substrate conversions. It offers the capability of dynamically altering nutrients, oxygenation, pH, carbon dioxide, and substrate concentration without disturbing the microgravity simulation, or Couette flow, of the reactor. All progeny of the original cell inoculum may be acclimated to the simulated microgravity in the absence of a substrate or nutrient. The reactor has the promise of allowing scientists to probe the long term effects of weightlessness on cell interactions in plants, bacteria, yeast, and fungi. The reactor is designed to have a flow field growth chamber with uniform shear stress, yet transfer

NMR system and method having a permanent magnet providing a rotating magnetic field

DOEpatents

Schlueter, Ross D [Berkeley, CA; Budinger, Thomas F [Berkeley, CA

2009-05-19

Disclosed herein are systems and methods for generating a rotating magnetic field. The rotating magnetic field can be used to obtain rotating-field NMR spectra, such as magic angle spinning spectra, without having to physically rotate the sample. This result allows magic angle spinning NMR to be conducted on biological samples such as live animals, including humans.

Anaerobic Membrane Bioreactor for Continuous Lactic Acid Fermentation

PubMed Central

Fan, Rong; Ebrahimi, Mehrdad; Czermak, Peter

2017-01-01

Membrane bioreactor systems can enhance anaerobic lactic acid fermentation by reducing product inhibition, thus increasing productivity. In batch fermentations, the bioconversion of glucose is strongly inhibited in the presence of more than 100 g·L−1 lactic acid and is only possible when the product is simultaneously removed, which can be achieved by ceramic membrane filtration. The crossflow velocity is a more important determinant of flux than the transmembrane pressure. Therefore, to stabilize the performance of the membrane bioreactor system during continuous fermentation, the crossflow velocity was controlled by varying the biomass concentration, which was monitored in real-time using an optical sensor. Continuous fermentation under these conditions, thus, achieved a stable productivity of ~8 g·L−1·h−1 and the concentration of lactic acid was maintained at ~40 g·L−1 at a dilution rate of 0.2 h−1. No residual sugar was detected in the steady state with a feed concentration of 50 g·L−1. PMID:28467384

Generation and physical characteristics of the ERTS MSS system corrected computer compatible tapes

NASA Technical Reports Server (NTRS)

Thomas, V. L.

1973-01-01

The generation and format are discussed of the ERTS system corrected multispectral scanner computer compatible tapes. The discussion includes spacecraft sensors, scene characteristics, data transmission, and conversion of data to computer compatible tapes at the NASA Data Processing Facility. Geometeric and radiometric corrections, tape formats, and the physical characteristics of the tapes are also included.

THz Dynamic Nuclear Polarization NMR

PubMed Central

Nanni, Emilio A.; Barnes, Alexander B.; Griffin, Robert G.; Temkin, Richard J.

2013-01-01

Dynamic nuclear polarization (DNP) increases the sensitivity of nuclear magnetic resonance (NMR) spectroscopy by using high frequency microwaves to transfer the polarization of the electrons to the nuclear spins. The enhancement in NMR sensitivity can amount to a factor of well above 100, enabling faster data acquisition and greatly improved NMR measurements. With the increasing magnetic fields (up to 23 T) used in NMR research, the required frequency for DNP falls into the THz band (140–600 GHz). Gyrotrons have been developed to meet the demanding specifications for DNP NMR, including power levels of tens of watts; frequency stability of a few megahertz; and power stability of 1% over runs that last for several days to weeks. Continuous gyrotron frequency tuning of over 1 GHz has also been demonstrated. The complete DNP NMR system must include a low loss transmission line; an optimized antenna; and a holder for efficient coupling of the THz radiation to the sample. This paper describes the DNP NMR process and illustrates the THz systems needed for this demanding spectroscopic application. THz DNP NMR is a rapidly developing, exciting area of THz science and technology. PMID:24639915

A self optimizing synthetic organic reactor system using real-time in-line NMR spectroscopy.

PubMed

Sans, Victor; Porwol, Luzian; Dragone, Vincenza; Cronin, Leroy

2015-02-01

A configurable platform for synthetic chemistry incorporating an in-line benchtop NMR that is capable of monitoring and controlling organic reactions in real-time is presented. The platform is controlled via a modular LabView software control system for the hardware, NMR, data analysis and feedback optimization. Using this platform we report the real-time advanced structural characterization of reaction mixtures, including 19 F, 13 C, DEPT, 2D NMR spectroscopy (COSY, HSQC and 19 F-COSY) for the first time. Finally, the potential of this technique is demonstrated through the optimization of a catalytic organic reaction in real-time, showing its applicability to self-optimizing systems using criteria such as stereoselectivity, multi-nuclear measurements or 2D correlations.

Farseer-NMR: automatic treatment, analysis and plotting of large, multi-variable NMR data.

PubMed

Teixeira, João M C; Skinner, Simon P; Arbesú, Miguel; Breeze, Alexander L; Pons, Miquel

2018-05-11

We present Farseer-NMR ( https://git.io/vAueU ), a software package to treat, evaluate and combine NMR spectroscopic data from sets of protein-derived peaklists covering a range of experimental conditions. The combined advances in NMR and molecular biology enable the study of complex biomolecular systems such as flexible proteins or large multibody complexes, which display a strong and functionally relevant response to their environmental conditions, e.g. the presence of ligands, site-directed mutations, post translational modifications, molecular crowders or the chemical composition of the solution. These advances have created a growing need to analyse those systems' responses to multiple variables. The combined analysis of NMR peaklists from large and multivariable datasets has become a new bottleneck in the NMR analysis pipeline, whereby information-rich NMR-derived parameters have to be manually generated, which can be tedious, repetitive and prone to human error, or even unfeasible for very large datasets. There is a persistent gap in the development and distribution of software focused on peaklist treatment, analysis and representation, and specifically able to handle large multivariable datasets, which are becoming more commonplace. In this regard, Farseer-NMR aims to close this longstanding gap in the automated NMR user pipeline and, altogether, reduce the time burden of analysis of large sets of peaklists from days/weeks to seconds/minutes. We have implemented some of the most common, as well as new, routines for calculation of NMR parameters and several publication-quality plotting templates to improve NMR data representation. Farseer-NMR has been written entirely in Python and its modular code base enables facile extension.

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.

Modeling and design of optimal flow perfusion bioreactors for tissue engineering applications.

PubMed

Hidalgo-Bastida, L Araida; Thirunavukkarasu, Sundaramoorthy; Griffiths, Sarah; Cartmell, Sarah H; Naire, Shailesh

2012-04-01

Perfusion bioreactors have been used in different tissue engineering applications because of their consistent distribution of nutrients and flow-induced shear stress within the tissue-engineering scaffold. A widely used configuration uses a scaffold with a circular cross-section enclosed within a cylindrical chamber and inlet and outlet pipes which are connected to the chamber on either side through which media is continuously circulated. However, fluid-flow experiments and simulations have shown that the majority of the flow perfuses through the center. This pattern creates stagnant zones in the peripheral regions as well as in those of high flow rate near the inlet and outlet. This non-uniformity of flow and shear stress, owing to a circular design, results in limited cell proliferation and differentiation in these areas. The focus of this communication is to design an optimized perfusion system using computational fluid dynamics as a mathematical tool to overcome the time-consuming trial and error experimental method. We compared the flow within a circular and a rectangular bioreactor system. Flow simulations within the rectangular bioreactor are shown to overcome the limitations in the circular design. This communication challenges the circular cross-section bioreactor configuration paradigm and provides proof of the advantages of the new design over the existing one. Copyright © 2011 Wiley Periodicals, Inc.

Tissue grown in NASA Bioreactor

NASA Technical Reports Server (NTRS)

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).

Membrane Distillation Bioreactor (MDBR) - A lower Green-House-Gas (GHG) option for industrial wastewater reclamation.

PubMed

Goh, Shuwen; Zhang, Jinsong; Liu, Yu; Fane, Anthony G

2015-12-01

A high-retention membrane bioreactor system, the Membrane Distillation Bioreactor (MDBR) is a wastewater reclamation process which has the potential to tap on waste heat generated in industries to produce high quality product water. There are a few key factors which could make MDBR an attractive advanced treatment option, namely tightening legal requirements due to increasing concerns on the micropollutants in industrial wastewater effluents as well as concerns over the electrical requirement of pressurized advanced treatment processes and greenhouse gas emissions associated with wastewater reclamation. This paper aims to provide a consolidated review on the current state of research for the MDBR system and to evaluate the system as a possible lower Green House Gas (GHG) emission option for wastewater reclamation using the membrane bioreactor-reverse osmosis (MBR-RO) system as a baseline for comparison. The areas for potential applications and possible configurations for MDBR applications are discussed. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

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.

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.

Bioreactors to Influence Stem Cell Fate: Augmentation of Mesenchymal Stem Cell Signaling Pathways via Dynamic Culture Systems

PubMed Central

Yeatts, Andrew B.; Choquette, Daniel T.; Fisher, John P.

2012-01-01

Background Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. Scope of Review This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. Major Conclusions The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. General Significance Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. PMID:22705676

Regulating Glucose and pH, and Monitoring Oxygen in a Bioreactor

NASA Technical Reports Server (NTRS)

Anderson, Melody M.; Pellis, Neat R.; Jeevarajan, Antony S.; Taylor, Thomas D.; Xu, Yuanhang; Gao, Frank

2006-01-01

A system that automatically regulates the concentration of glucose or pH in a liquid culture medium that is circulated through a rotating-wall perfused bioreactor is described. Another system monitors the concentration of oxygen in the culture medium.

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.

Performance of a novel baffled osmotic membrane bioreactor-microfiltration hybrid system under continuous operation for simultaneous nutrient removal and mitigation of brine discharge.

PubMed

Pathak, Nirenkumar; Chekli, Laura; Wang, Jin; Kim, Youngjin; Phuntsho, Sherub; Li, Sheng; Ghaffour, Noreddine; Leiknes, TorOve; Shon, Hokyong

2017-09-01

The present study investigated the performance of an integrated osmotic and microfiltration membrane bioreactor system for wastewater treatment employing baffles in the reactor. Thus, this reactor design enables both aerobic and anoxic processes in an attempt to reduce the process footprint and energy costs associated with continuous aeration. The process performance was evaluated in terms of water flux, salinity build up in the bioreactor, organic and nutrient removal and microbial activity using synthetic reverse osmosis (RO) brine as draw solution (DS). The incorporation of MF membrane was effective in maintaining a reasonable salinity level (612-1434mg/L) in the reactor which resulted in a much lower flux decline (i.e. 11.48-6.98LMH) as compared to previous studies. The stable operation of the osmotic membrane bioreactor-forward osmosis (OMBR-FO) process resulted in an effective removal of both organic matter (97.84%) and nutrient (phosphate 87.36% and total nitrogen 94.28%), respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

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 concepts for bioreactors in space

NASA Technical Reports Server (NTRS)

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

1987-01-01

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. 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 spacecraft, space stations and other extra-terrestrial habitats.

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.

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.

Scale up of fuel ethanol production from sugar beet juice using loofa sponge immobilized bioreactor.

PubMed

Ogbonna, J C; Mashima, H; Tanaka, H

2001-01-01

Production of fuel ethanol from sugar beet juice, using cells immobilized on loofa sponge was investigated. Based on ethanol productivity and ease of cell immobilization, a flocculating yeast strain, Saccharomyces cerevisiae IR2 was selected for ethanol production from sugar beet juice. It was found that raw sugar beet juice was an optimal substrate for ethanol production, requiring neither pH adjustment nor nitrogen source supplement. When compared with a 2 l bubble column bioreactor, mixing was not sufficient in an 8 l bioreactor containing a bed of sliced loofa sponges and consequently, the immobilized cells were not uniformly distributed within the bed. Most of the cells were immobilized in the lower part of the bed and this resulted in decreased ethanol productivity. By using an external loop bioreactor, constructing the fixed bed with cylindrical loofa sponges, dividing the bed into upper, middle and lower sections with approximately 1 cm spaces between them and circulating the broth through the loop during the immobilization, uniform cell distribution within the bed was achieved. Using this method, the system was scaled up to 50 l and when compared with the 2 l bubble column bioreactor, there were no significant differences (P > 0.05) in ethanol productivity and yield. By using external loop bioreactor to immobilize the cells uniformly on the loofa sponge beds, efficient large scale ethanol production systems can be constructed.

LTCC based bioreactors for cell cultivation

NASA Astrophysics Data System (ADS)

Bartsch, H.; Welker, T.; Welker, K.; Witte, H.; Müller, J.

2016-01-01

LTCC multilayers offer a wide range of structural options and flexibility of connections not available in standard thin film technology. Therefore they are considered as material base for cell culture reactors. The integration of microfluidic handling systems and features for optical and electrical capturing of indicators for cell culture growth offers the platform for an open system concept. The present paper assesses different approaches for the creation of microfluidic channels in LTCC multilayers. Basic functions required for the fluid management in bioreactors include temperature and flow control. Both features can be realized with integrated heaters and temperature sensors in LTCC multilayers. Technological conditions for the integration of such elements into bioreactors are analysed. The temperature regulation for the system makes use of NTC thermistor sensors which serve as real value input for the control of the heater. It allows the adjustment of the fluid temperature with an accuracy of 0.2 K. The tempered fluid flows through the cell culture chamber. Inside of this chamber a thick film electrode array monitors the impedance as an indicator for the growth process of 3-dimensional cell cultures. At the system output a flow sensor is arranged to monitor the continual flow. For this purpose a calorimetric sensor is implemented, and its crucial design parameters are discussed. Thus, the work presented gives an overview on the current status of LTCC based fluid management for cell culture reactors, which provides a promising base for the automation of cell culture processes.

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

Comparison of Leachate Quality from Aerobic and Anaerobic Municipal Solid Waste Bioreactors

NASA Astrophysics Data System (ADS)

Borglin, S. E.; Hazen, T. C.; Oldenburg, C. M.

2002-12-01

Municipal solid waste landfills are becoming a drain on the resources of local municipalities as the requirements for stabilization and containment become increasingly stringent. Current regulations limit the moisture in the landfill to minimize leachate production and lower the potential for release of leachate to the environment. Recent research has shown that addition and recycling of moisture in the waste optimizes the biodegradation of stabilization and also provides a means for leachate treatment. This study compares the characteristics of leachate produced from aerobic and anaerobic laboratory bioreactors, and leachate collected from a full-scale anaerobic bioreactor. The laboratory reactors consisted of 200-liter tanks filled with fresh waste materials with the following conditions: (a) aerobic (air injection with leachate recirculation), (b) anaerobic (leachate recirculation). The leachate from the reactors was monitored for metals, nutrients, organic carbon, and microbiological activity for up to 500 days. Leachate from the aerobic tank had significantly lower concentrations of all potential contaminants, both organic and metal, after only a few weeks of operation. Metals leaching was low throughout the test period for the aerobic tanks, and decreased over time for the anaerobic tanks. Organic carbon as measured by BOD, COD, TOC, and COD were an order of magnitude higher in the leachate from the anaerobic system. Microbiological assessment by lipid analysis, enzyme activity assays, and cell counts showed high biomass and diversity in both the aerobic and anaerobic bioreactors, with higher activity in the anaerobic leachate. Results from the full-scale anaerobic bioreactor were not significantly different from those of the laboratory anaerobic bioreactor. The reduction in noxious odors was a significant advantage of the aerobic system. These results suggest that aerobic management of landfills could reduce or eliminate the need for leachate treatment

Bioreactor Engineering of Stem Cell Environments

PubMed Central

Tandon, Nina; Marolt, Darja; Cimetta, Elisa; Vunjak-Novakovic, Gordana

2013-01-01

Stem cells hold promise to revolutionize modern medicine by development of new therapies, disease models and drug screening systems. Standard cell culture systems have limited biological relevance because they do not recapitulate the complex 3-dimensional interactions and biophysical cues that characterize the in vivo environment. In this review, we discuss the current advances in engineering stem cell environments using novel biomaterials and bioreactor technologies. We also reflect on the challenges the field is currently facing with regard to translation of stem cell based therapies into the clinic. PMID:23531529

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.

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.

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

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.

Membrane filtration device for studying compression of fouling layers in membrane bioreactors

PubMed Central

Bugge, Thomas Vistisen; Larsen, Poul; Nielsen, Per Halkjær; Christensen, Morten Lykkegaard

2017-01-01

A filtration devise was developed to assess compressibility of fouling layers in membrane bioreactors. The system consists of a flat sheet membrane with air scouring operated at constant transmembrane pressure to assess the influence of pressure on resistance of fouling layers. By fitting a mathematical model, three model parameters were obtained; a back transport parameter describing the kinetics of fouling layer formation, a specific fouling layer resistance, and a compressibility parameter. This stands out from other on-site filterability tests as model parameters to simulate filtration performance are obtained together with a characterization of compressibility. Tests on membrane bioreactor sludge showed high reproducibility. The methodology’s ability to assess compressibility was tested by filtrations of sludges from membrane bioreactors and conventional activated sludge wastewater treatment plants from three different sites. These proved that membrane bioreactor sludge showed higher compressibility than conventional activated sludge. In addition, detailed information on the underlying mechanisms of the difference in fouling propensity were obtained, as conventional activated sludge showed slower fouling formation, lower specific resistance and lower compressibility of fouling layers, which is explained by a higher degree of flocculation. PMID:28749990

High cell density cultivation and recombinant protein production with Escherichia coli in a rocking-motion-type bioreactor

PubMed Central

2010-01-01

Background Single-use rocking-motion-type bag bioreactors provide advantages compared to standard stirred tank bioreactors by decreased contamination risks, reduction of cleaning and sterilization time, lower investment costs, and simple and cheaper validation. Currently, they are widely used for cell cultures although their use for small and medium scale production of recombinant proteins with microbial hosts might be very attractive. However, the utilization of rocking- or wave-induced motion-type bioreactors for fast growing aerobic microbes is limited because of their lower oxygen mass transfer rate. A conventional approach to reduce the oxygen demand of a culture is the fed-batch technology. New developments, such as the BIOSTAT® CultiBag RM system pave the way for applying advanced fed-batch control strategies also in rocking-motion-type bioreactors. Alternatively, internal substrate delivery systems such as EnBase® Flo provide an opportunity for adopting simple to use fed-batch-type strategies to shaken cultures. Here, we investigate the possibilities which both strategies offer in view of high cell density cultivation of E. coli and recombinant protein production. Results Cultivation of E. coli in the BIOSTAT® CultiBag RM system in a conventional batch mode without control yielded an optical density (OD600) of 3 to 4 which is comparable to shake flasks. The culture runs into oxygen limitation. In a glucose limited fed-batch culture with an exponential feed and oxygen pulsing, the culture grew fully aerobically to an OD600 of 60 (20 g L-1 cell dry weight). By the use of an internal controlled glucose delivery system, EnBase® Flo, OD600 of 30 (10 g L-1 cell dry weight) is obtained without the demand of computer controlled external nutrient supply. EnBase® Flo also worked well in the CultiBag RM system with a recombinant E. coli RB791 strain expressing a heterologous alcohol dehydrogenase (ADH) to very high levels, indicating that the enzyme based feed

Yarrowia lipolytica morphological mutant enables lasting in situ immobilization in bioreactor.

PubMed

Vandermies, Marie; Kar, Tambi; Carly, Frédéric; Nicaud, Jean-Marc; Delvigne, Frank; Fickers, Patrick

2018-04-26

In the present study, we have isolated and characterized a Yarrowia lipolytica morphological mutant growing exclusively in the pseudohyphal morphology. The gene responsible for this phenotype, YALI0E06519g, was identified as homologous to the mitosis regulation gene HSL1 from Saccharomyces cerevisiae. Taking advantage of its morphology, we achieved the immobilization of the Δhsl1 mutant on the metallic structured packing of immobilized-cell bioreactors. We obtained significant cell retention and growth on the support during shake flask and bioreactor experiments without an attachment step prior to the culture. The system of medium aspersion on the packing ensured oxygen availability in the absence of agitation and minimized the potential release of cells in the culture medium. Additionally, the metallic packing proved its facility of cleaning and sterilization after fermentation. This combined use of morphological mutation and bioreactor design is a promising strategy to develop continuous processes for the production of recombinant protein and metabolites using Y. lipolytica. Graphical Abstract.

Corn forage biological pretreatment by Trametes versicolor in a tray bioreactor.

PubMed

Planinić, Mirela; Zelić, Bruno; Čubel, Ivan; Bucić-Kojić, Ana; Tišma, Marina

2016-08-01

Trametes versicolor is a white-rot fungus known to be efficient in lignin removal due to its complex extracellular lignocellulolytic enzymatic system. Therefore, it can be used in the treatment of lignocellulose waste from agro, food, and wood industries. In a first experiment, corn forage treatment with T. versicolor was investigated in laboratory jars. In a second experiment, the process was scaled up to a tray bioreactor. In the tray bioreactor, the process of lignin degradation was improved, resulting in an increase in lignin conversion of up to 71% during seven days' treatment. © The Author(s) 2016.

Design of well and groove microchannel bioreactors for cell culture.

PubMed

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

2009-03-01

Microfluidic bioreactors have been shown valuable for various cellular applications. The use of micro-wells/grooves bioreactors, in which micro-topographical features are used to protect sensitive cells from the detrimental effects of fluidic shear stress, is a promising approach to culture sensitive cells in these perfusion microsystems. However, such devices exhibit substantially different fluid dynamics and mass transport characteristics compared to conventional planar microchannel reactors. In order to properly design and optimize these systems, fluid and mass transport issues playing a key role in microscale bioreactors should be adequately addressed. The present work is a parametric study of micro-groove/micro-well microchannel bioreactors. Operation conditions and design parameters were theoretically examined via a numerical model. The complex flow pattern obtained at grooves of various depths was studied and the shear protection factor compared to planar microchannels was evaluated. 3D flow simulations were preformed in order to examine the shear protection factor in micro-wells, which were found to have similar attributes as the grooves. The oxygen mass transport problem, which is coupled to the fluid mechanics problem, was solved for various groove geometries and for several cell types, assuming a defined shear stress limitation. It is shown that by optimizing the groove depth, the groove bioreactor may be used to effectively maximize the number of cells cultured within it or to minimize the oxygen gradient existing in such devices. Moreover, for sensitive cells having a high oxygen demand (e.g., hepatocytes) or low endurance to shear (e.g., human embryonic stem cells), results show that the use of grooves is an enabling technology, since under the same physical conditions the cells cannot be cultured for long periods of time in a planar microchannel. In addition to the theoretical model findings, the culture of human foreskin fibroblasts in groove (30

NMR transmit-receive system with short recovery time and effective isolation

NASA Astrophysics Data System (ADS)

Jurga, K.; Reynhardt, E. C.; Jurga, S.

A transmit-receive system with a short recovery time and excellent isolation has been developed. The system operates in conjunction with an ENI Model 3200L broadband amplifier and a spin-lock NMR pulse spectrometer. The system has been tested in the frequency range 5.5 to 52 MHz and seems not to generate any background noise.

Mathematical modeling of continuous ethanol fermentation in a membrane bioreactor by pervaporation compared to conventional system: Genetic algorithm.

PubMed

Esfahanian, Mehri; Shokuhi Rad, Ali; Khoshhal, Saeed; Najafpour, Ghasem; Asghari, Behnam

2016-07-01

In this paper, genetic algorithm was used to investigate mathematical modeling of ethanol fermentation in a continuous conventional bioreactor (CCBR) and a continuous membrane bioreactor (CMBR) by ethanol permselective polydimethylsiloxane (PDMS) membrane. A lab scale CMBR with medium glucose concentration of 100gL(-1) and Saccharomyces cerevisiae microorganism was designed and fabricated. At dilution rate of 0.14h(-1), maximum specific cell growth rate and productivity of 0.27h(-1) and 6.49gL(-1)h(-1) were respectively found in CMBR. However, at very high dilution rate, the performance of CMBR was quite similar to conventional fermentation on account of insufficient incubation time. In both systems, genetic algorithm modeling of cell growth, ethanol production and glucose concentration were conducted based on Monod and Moser kinetic models during each retention time at unsteady condition. The results showed that Moser kinetic model was more satisfactory and desirable than Monod model. Copyright © 2016 Elsevier Ltd. All rights reserved.

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.

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...

76 FR 66666 - Basic Service Tier Encryption Compatibility Between Cable Systems and Consumer Electronics Equipment

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-27

... 11-153] Basic Service Tier Encryption Compatibility Between Cable Systems and Consumer Electronics... substantially affect compatibility between cable service and consumer electronics equipment for most subscribers... problems between cable service and consumer electronics equipment were limiting and/or precluding the...

Aircraft Electromagnetic Compatibility.

DTIC Science & Technology

1987-06-01

Human Exposure to Radio Frequency Electromagnetic Fields , 300 KiloHertz to 100 GigaHertz." 6. ARINC 429-8, "Digital Information Transfer System (DITS...142 V EXECUTIVE SUMMARY The Aircraft Electromagnetic Compatibility guidelines document deals with electromagnetic compatibility in a... electromagnetic interference paths (figure EI. TYPE PATH 400 Hz Electrostatic MagneticCharge Electric Field Transients 5 R d t Coupling 150/i 300o Wire

NMR at Low and Ultra-Low Temperatures

PubMed Central

Tycko, Robert

2017-01-01

Conspectus Solid state nuclear magnetic resonance (NMR) measurements at low temperatures have been common in physical sciences for many years, and are becoming increasingly important in studies of biomolecular systems. This article reviews a diverse set of projects from my laboratory, dating back to the early 1990s, that illustrate the motivations for low-temperature solid state NMR, the types of information that are available from the measurements, and likely directions for future research. These projects include NMR studies of both physical and biological systems, performed at low (cooled with nitrogen, down to 77 K) and very low (cooled with helium, below 77 K) temperatures, and performed with and without magic-angle spinning (MAS). In NMR studies of physical systems, the main motivation is to study phenomena that occur only at low temperatures. Two examples from my laboratory are studies of molecular rotation and an orientational ordering in solid C60 at low temperatures and studies of unusual electronic states, called skyrmions, in two-dimensionally confined electron systems within semiconductor quantum wells. NMR measurements on quantum wells were facilitated by optical pumping of nuclear spin polarizations, a signal enhancement phenomenon that exists at very low temperatures. In studies of biomolecular systems, motivations for low-temperature NMR include suppression of molecular tumbling (thereby permitting solid state NMR measurements on soluble proteins), suppression of conformational exchange (thereby permitting quantitation of conformational distributions), and trapping of transient intermediate states in a non-equilibrium kinetic process (by rapid freeze-quenching). Solid state NMR measurements on AIDS-related peptide/antibody complexes, chemically denatured states of the model protein HP35, and a transient intermediate in the rapid folding pathway of HP35 illustrate these motivations. NMR sensitivity generally increases with decreasing sample

Utilization of microgravity bioreactors for differentiation of mammalian skeletal tissue

NASA Technical Reports Server (NTRS)

Klement, B. J.; Spooner, B. S.

1993-01-01

Bioreactor cell and tissue culture vessels can be used to study bone development in a simulated microgravity environment. These vessels will also provide an advantageous, low maintenance culture system on space station Freedom. Although many types of cells and tissues can potentially utilize this system, our particular interest is in developing bone tissue. We have characterized an organ culture system utilizing embryonic mouse pre-metatarsal mesenchyme, documenting morphogenesis and differentiation as cartilage rods are formed, with subsequent terminal chondrocyte differentiation to hypertrophied cells. Further development to form bone tissue is achieved by supplementation of the culture medium. Research using pre-metatarsal tissue, combined with the bioreactor culture hardware, could give insight into the advantages and/or disadvantages of conditions experienced in microgravity. Studies such as these have the potential to enhance understanding of bone development and adult bone physiology, and may help define the processes of bone demineralization experienced in space and in pathological conditions here on earth.

Shear stress enhances microcin B17 production in a rotating wall bioreactor, but ethanol stress does not

NASA Technical Reports Server (NTRS)

Gao, Q.; Fang, A.; Pierson, D. L.; Mishra, S. K.; Demain, A. L.

2001-01-01

Stress, including that caused by ethanol, has been shown to induce or promote secondary metabolism in a number of microbial systems. Rotating-wall bioreactors provide a low stress and simulated microgravity environment which, however, supports only poor production of microcin B17 by Escherichia coli ZK650, as compared to production in agitated flasks. We wondered whether the poor production is due to the low level of stress and whether increasing stress in the bioreactors would raise the amount of microcin B17 formed. We found that applying shear stress by addition of a single Teflon bead to a rotating wall bioreactor improved microcin B17 production. By contrast, addition of various concentrations of ethanol to such bioreactors (or to shaken flasks) failed to increase microcin B17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the bioreactor were much more resistant to the growth-inhibitory and production-inhibitory effects of ethanol than cells growing in shaken flasks.

Compatible ionic liquid-cellulases system for hydrolysis of lignocellulosic biomass.

PubMed

Wang, Ying; Radosevich, Mark; Hayes, Douglas; Labbé, Nicole

2011-05-01

Ionic liquids (ILs) have been increasingly recognized as novel solvents for dissolution and pretreatment of cellulose. However, cellulases are inactivated in the presence of ILs, even when present at low concentrations. To more fully exploit the benefits of ILs it is critical to develop a compatible IL-cellulases system in which the IL is able to effectively solubilize and activate the lignocellulosic biomass, and the cellulases possess high stability and activity. In this study, we investigated the stability and activity of a commercially available cellulases mixture in the presence of different concentrations of 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]). A mixture of cellulases and β-glucosidase (Celluclast1.5L, from Trichoderma reesei, and Novozyme188, from Aspergillus niger, respectively) retained 77% and 65% of its original activity after being pre-incubated in 15% and 20% (w/v) IL solutions, respectively, at 50°C for 3 h. The cellulases mixture also retained high activity in 15% [Emim][OAc] to hydrolyze Avicel, a model substrate for cellulose analysis, with conversion efficiency of approximately 91%. Notably, the presence of different amounts of yellow poplar lignin did not interfere significantly with the enzymatic hydrolysis of Avicel. Using this IL-cellulase system (15% [Emim][OAc]), the saccharification of yellow poplar biomass was also significantly improved (33%) compared to the untreated control (3%) during the first hour of enzymatic hydrolysis. Together, these findings provide compelling evidence that [Emim][OAc] was compatible with the cellulase mixture, and this compatible IL-cellulases system is promising for efficient activation and hydrolysis of native biomass to produce biofuels and co-products from the individual biomass components. Copyright © 2010 Wiley Periodicals, Inc.

Partitioning of nitroxides in dispersed systems investigated by ultrafiltration, EPR and NMR spectroscopy.

PubMed

Krudopp, Heimke; Sönnichsen, Frank D; Steffen-Heins, Anja

2015-08-15

The partitioning behavior of paramagnetic nitroxides in dispersed systems can be determined by deconvolution of electron paramagnetic resonance (EPR) spectra giving equivalent results with the validated methods of ultrafiltration techniques (UF) and pulsed-field gradient nuclear magnetic resonance spectroscopy (PFG-NMR). The partitioning behavior of nitroxides with increasing lipophilicity was investigated in anionic, cationic and nonionic micellar systems and 10 wt% o/w emulsions. Apart from EPR spectra deconvolution, the PFG-NMR was used in micellar solutions as a non-destructive approach, while UF based on separation of very small volume of the aqueous phase. As a function of their substituent and lipophilicity, the proportions of nitroxides that were solubilized in the micellar or emulsion interface increased with increasing nitroxide lipophilicity for all emulsifier used. Comparing the different approaches, EPR deconvolution and UF revealed comparable nitroxide proportions that were solubilized in the interfaces. Those proportions were higher than found with PFG-NMR. For PFG-NMR self-diffusion experiments the reduced nitroxides were used revealing a high dynamic of hydroxylamines and emulsifiers. Deconvolution of EPR spectra turned out to be the preferred method for measuring the partitioning behavior of paramagnetic molecules as it enables distinguishing between several populations at their individual solubilization sites. Copyright © 2015 Elsevier Inc. All rights reserved.

Manipulating and probing the polarisation of a methyl tunnelling system by field-cycling NMR

NASA Astrophysics Data System (ADS)

Zhang, Bo; Abu-Khumra, Sabah M. M.; Aibout, Abdellah; Horsewill, Anthony J.

2017-02-01

In NMR the polarisation of the Zeeman system may be routinely probed and manipulated by applying resonant rf pulses. As with spin-1/2 nuclei, at low temperature the quantum tunnelling states of a methyl rotor are characterised by two energy levels and it is interesting to consider how these tunnelling states might be probed and manipulated in an analogous way to nuclear spins in NMR. In this paper experimental procedures based on magnetic field-cycling NMR are described where, by irradiating methyl tunnelling sidebands, the polarisations of the methyl tunnelling systems are measured and manipulated in a prescribed fashion. At the heart of the technique is a phenomenon that is closely analogous to dynamic nuclear polarisation and the solid effect where forbidden transitions mediate polarisation transfer between 1H Zeeman and methyl tunnelling systems. Depending on the irradiated sideband, both positive and negative polarisations of the tunnelling system are achieved, the latter corresponding to population inversion and negative tunnelling temperatures. The transition mechanics are investigated through a series of experiments and a theoretical model is presented that provides good quantitative agreement.

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.

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.

Correlation between mass transfer coefficient kLa and relevant operating parameters in cylindrical disposable shaken bioreactors on a bench-to-pilot scale.

PubMed

Klöckner, Wolf; Gacem, Riad; Anderlei, Tibor; Raven, Nicole; Schillberg, Stefan; Lattermann, Clemens; Büchs, Jochen

2013-12-02

Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen kLa and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal kLa correlation. A Respiration Activity Monitoring System (RAMOS) was used to measure kLa values in cylindrical disposable shaken bioreactors and Buckingham's π-Theorem was applied to define a dimensionless equation for kLa. In this way, a scale- and volume-independent kLa correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures. The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/- 30%. To our knowledge, this is the first kLa correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale.

Weapon and Sighting System Compatibility Assessment for Prototype Maxillofacial Protection Devices

DTIC Science & Technology

2013-03-01

No Maxillofacial Protection, No Visor (figure 38) ...........................22 7.2 Sighting System Compatibility – Bradley Fighting Vehicle ( BFV ...23 Figure 39. FASTBAC, bracket only ( BFV ...23 Figure 40. FASTBAC bracket and maxillofacial protection ( BFV ). ............................................24 Figure 41. FIREBALL

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

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.

Comparison of online reporting systems and their compatibility check with respective adverse drug reaction reporting forms.

PubMed

Maharshi, Vikas; Nagar, Pravesh

2017-01-01

Different forms and online tools are available in different countries for spontaneous reporting, one of the most widely used methods of pharmacovigilance. Capturing sufficient information and adequate compatibility of online systems with respective reporting form is highly desirable for appropriate reporting of adverse drug reactions (ADRs). This study was aimed to compare three major online reporting systems (US, UK, and WHO) of the world and also to check their compatibility with the respective ADR reporting form. A total of 89 data elements to provide relevant information were found out from above three online reporting systems. All three online systems were compared regarding magnitude of information captured by each of them and scoring was done by providing a score of "1" to each element. Compatibility of ADR reporting forms of India (Red form), US (Form 3500), and UK (Yellow card form) was assessed by comparing the information gathered by them with that can be entered into their respective online reporting systems, namely, "VigiFlow," "US online reporting," and "Yellow card online reporting." Each unmatching item was given a score of "-1". VigiFlow scored "74" points, whereas online reporting systems of the US and UK scored "56" and "49," respectively, regarding magnitude of the information gathered by them. Compatibility score was found to be "0," "-9," and "-26" in case of ADR reporting systems of US, UK, and India, respectively. Our study reveals that "VigiFlow" is capable of capturing the maximum amount of information but "Form 3500" and "Online reporting system of US" are maximally compatible to each other among ADR reporting systems of all three countries.

Development of a bioreactor system for the decolorization of dyes by Coriolus versicolor f. antarcticus.

PubMed

Diorio, L A; Mercuri, A A; Nahabedian, D E; Forchiassin, F

2008-05-01

Decolorization of 100 microM malachite green (MG) by Coriolus versicolor f. antarcticus using a two-phase bioreactor, was investigated. In the first phase the decolorization ability of this fungus, growing under conditions of solid-state fermentation (SSF), was proved; in the second phase the capacity of the enzymes present in extracts from the solid residues was exploited. During the first phase using the same culture in the bioreactor, five consecutive charges were made, each with 75 ml of 100 microM MG solution, at 28 degrees C. Each cycle ended when MG solution reached a decolorization of 50%, at this time the bioreactor was discharged to a stainless steel coil at 50 degrees C, initiating the second phase of decolorization. Time required in order to reach 50% decolorization during the first phase varied between 25 and 65 min, with an average retention time of 48 min. The second stage had a retention time of 120 min. Residual MG after this phase varied from 0% to 6.3%. The role of laccase and Mn-peroxidase in MG decolorization is discussed. Toxicity of MG solutions before and after decolorization treatments was assayed using Lumbriculus variegatus as test organism.

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.

Permitting of Landfill Bioreactor Operations: Ten Years after ...

EPA Pesticide Factsheets

Prior to promulgation of the Rule, there were approximately 20 full-scale bioreactor projects in North America, including one in Canada. Of these, six were permitted by EPA (four Project XL sites and two projects listed separately under a cooperative research agreement at the Outer Loop Landfill in Kentucky). In March 2014, there were about 40 bioreactor projects reported, including 30 active RD&D projects in 11 approved states and one project on tribal lands. Wisconsin features the largest number of projects at 13, due primarily to the fact that landfill owners in the state must either eliminate landfill disposal of biodegradable materials or to achieve the complete stabilization of deposited organic waste at MSW landfills within 40 years after closure. Most landfill operators have selected a bioreactor approach to attempt to achieve the latter goal. In summary, only 16 of 50 (32%) states have currently adopted the Rule, meaning that development of RD&D permitting procedures that are consistent with EPA’s requirements has generally not occurred. The predominant single reason cited for not adopting the Rule was lack of interest amongst landfill facilities in the state. Subtitle D and its state derivatives already allow leachate recirculation over prescriptive (i.e., minimum technology) liner systems, which is often the primary goal of site operators seeking to control leachate treatment costs. Other reasons related to concerns over increased time, cost

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.

An MR-compatible gyroscope-based arm movement tracking system.

PubMed

Shirinbayan, S Iman; Rieger, Jochem W

2017-03-15

Functional magnetic resonance imaging is well suited to link neural population activation with movement parameters of complex natural arm movements. However, currently existing MR-compatible arm tracking devices are not constructed to measure arm joint movement parameters of unrestricted movements. Therefore, to date most research focuses on simple arm movements or includes very little knowledge about the actual movement kinematics. We developed a low cost gyroscope-based arm movement tracking system (GAMTS) that features MR-compatibility. The system consists of dual-axis analogue gyroscopes that measure rotations of upper and lower arm joints. After MR artifact reduction, the rotation angles of the individual arm joints are calculated and used to animate a realistic arm model that is implemented in the OpenSim platform. The OpenSim platform can then provide the kinematics of any point on the arm model. In order to demonstrate the capabilities of the system, we first assessed the quality of reconstructed wrist movements in a low-noise environment where typical MR-related problems are absent and finally, we validated the reconstruction in the MR environment. The system provides the kinematics of the whole arm when natural unrestricted arm movements are performed inside the MR-scanner. The GAMTS is reliably capable of reconstructing the kinematics of trajectories and the reconstruction error is small in comparison with the movement induced variation of speed, displacement, and rotation. Moreover, the system can be used to probe brain areas for their correlation with movement kinematics. Copyright © 2017 Elsevier B.V. All rights reserved.

Feasibility study of contaminant detection for food with ULF-NMR/MRI system using HTS-SQUID

NASA Astrophysics Data System (ADS)

Hatsukade, Yoshimi; Tsunaki, Shingo; Yamamoto, Masaaki; Abe, Takayuki; Hatta, Junichi; Tanaka, Saburo

2013-11-01

We have developed an ultra-low frequency (ULF) nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) system utilizing an HTS-SQUID for an application of contaminant detection in food and drink. In the system, a permanent magnet of 1.1 T was used to pre-polarize protons in a water sample. We measured NMR signals from water samples with or without various contaminants, such as stainless steel (SUS304), aluminum, and glass balls using the system. In the case that the contaminant was the SUS304 ball, the NMR signal intensity was reduced compared to that from the sample without the contaminant due to the remnant field of the contaminant. One-dimensional (1D) MRIs of the samples were also acquired to detect non-magnetic contaminants. In the 1D MRIs, changes of the MRI spectra were detected, corresponding to positions of the contaminants. These results show that the feasibility of the system to detect various contaminants in foods.

Electromagnetic Compatibility Design of the Computer Circuits

NASA Astrophysics Data System (ADS)

Zitai, Hong

2018-02-01

Computers and the Internet have gradually penetrated into every aspect of people’s daily work. But with the improvement of electronic equipment as well as electrical system, the electromagnetic environment becomes much more complex. Electromagnetic interference has become an important factor to hinder the normal operation of electronic equipment. In order to analyse the computer circuit compatible with the electromagnetic compatibility, this paper starts from the computer electromagnetic and the conception of electromagnetic compatibility. And then, through the analysis of the main circuit and system of computer electromagnetic compatibility problems, we can design the computer circuits in term of electromagnetic compatibility. Finally, the basic contents and methods of EMC test are expounded in order to ensure the electromagnetic compatibility of equipment.

Bioreactor engineering of stem cell environments.

PubMed

Tandon, Nina; Marolt, Darja; Cimetta, Elisa; Vunjak-Novakovic, Gordana

2013-11-15

Stem cells hold promise to revolutionize modern medicine by the development of new therapies, disease models and drug screening systems. Standard cell culture systems have limited biological relevance because they do not recapitulate the complex 3-dimensional interactions and biophysical cues that characterize the in vivo environment. In this review, we discuss the current advances in engineering stem cell environments using novel biomaterials and bioreactor technologies. We also reflect on the challenges the field is currently facing with regard to the translation of stem cell based therapies into the clinic. Copyright © 2013 Elsevier Inc. All rights reserved.

Bacterial Community Dynamics in Full-Scale Activated Sludge Bioreactors: Operational and Ecological Factors Driving Community Assembly and Performance

PubMed Central

Valentín-Vargas, Alexis; Toro-Labrador, Gladys; Massol-Deyá, Arturo A.

2012-01-01

The assembling of bacterial communities in conventional activated sludge (CAS) bioreactors was thought, until recently, to be chaotic and mostly unpredictable. Studies done over the last decade have shown that specific, and often, predictable random and non-random factors could be responsible for that process. These studies have also motivated a “structure–function” paradigm that is yet to be resolved. Thus, elucidating the factors that affect community assembly in the bioreactors is necessary for predicting fluctuations in community structure and function. For this study activated sludge samples were collected during a one-year period from two geographically distant CAS bioreactors of different size. Combining community fingerprinting analysis and operational parameters data with a robust statistical analysis, we aimed to identify relevant links between system performance and bacterial community diversity and dynamics. In addition to revealing a significant β-diversity between the bioreactors’ communities, results showed that the largest bioreactor had a less dynamic but more efficient and diverse bacterial community throughout the study. The statistical analysis also suggests that deterministic factors, as opposed to stochastic factors, may have a bigger impact on the community structure in the largest bioreactor. Furthermore, the community seems to rely mainly on mechanisms of resistance and functional redundancy to maintain functional stability. We suggest that the ecological theories behind the Island Biogeography model and the species-area relationship were appropriate to predict the assembly of bacterial communities in these CAS bioreactors. These results are of great importance for engineers and ecologists as they reveal critical aspects of CAS systems that could be applied towards improving bioreactor design and operation. PMID:22880016

Design and Fabrication of an MRI-Compatible, Autonomous Incubation System.

PubMed

Khalilzad-Sharghi, Vahid; Xu, Huihui

2015-10-01

Tissue engineers have long sought access to an autonomous, imaging-compatible tissue incubation system that, with minimum operator handling, can provide real-time visualization and quantification of cells, tissue constructs, and organs. This type of screening system, capable of operating noninvasively to validate tissue, can overcome current limitations like temperature shock, unsustainable cellular environments, sample contamination, and handling/stress. However, this type of system has been a major challenge, until now. Here, we describe the design, fabrication, and characterization of an innovative, autonomous incubation system that is compatible with a 9.4 T magnetic resonance imaging (MRI) scanner. Termed the e-incubator (patent pending; application number: 13/953,984), this microcontroller-based system is integrated into an MRI scanner and noninvasively screens cells and tissue cultures in an environment where temperature, pH, and media/gas handling are regulated. The 4-week study discussed herein details the continuous operation of the e-incubator for a tissue-engineered osteogenic construct, validated by LIVE/DEAD(®) cell assays and histology. The evolving MR quantitative parameters of the osteogenic construct were used as biomarkers for bone tissue engineering and to further validate the quality of the product noninvasively before harvesting. Importantly, the e-incubator reliably facilitates culturing cells and tissue constructs to create engineered tissues and/or investigate disease therapies.

Role of Bioreactor Technology in Tissue Engineering for Clinical Use and Therapeutic Target Design.

PubMed

Selden, Clare; Fuller, Barry

2018-04-24

Micro and small bioreactors are well described for use in bioprocess development in pre-production manufacture, using ultra-scale down and microfluidic methodology. However, the use of bioreactors to understand normal and pathophysiology by definition must be very different, and the constraints of the physiological environment influence such bioreactor design. This review considers the key elements necessary to enable bioreactors to address three main areas associated with biological systems. All entail recreation of the in vivo cell niche as faithfully as possible, so that they may be used to study molecular and cellular changes in normal physiology, with a view to creating tissue-engineered grafts for clinical use; understanding the pathophysiology of disease at the molecular level; defining possible therapeutic targets; and enabling appropriate pharmaceutical testing on a truly representative organoid, thus enabling better drug design, and simultaneously creating the potential to reduce the numbers of animals in research. The premise explored is that not only cellular signalling cues, but also mechano-transduction from mechanical cues, play an important role.

Effect of different fermentation strategies on β-mannanase production in fed-batch bioreactor system.

PubMed

Germec, Mustafa; Yatmaz, Ercan; Karahalil, Ercan; Turhan, İrfan

2017-05-01

Mannanases, one of the important enzyme group for industry, are produced by numerous filamentous fungi, especially Aspergillus species with different fermentation methods. The aim of this study was to show the best fermentation method of β-mannanase production for fungal growth in fermenter. Therefore, different fermentation strategies in fed-batch fermentation (suspended, immobilized cell, biofilm and microparticle-enhanced bioreactor) were applied for β-mannanase production from glucose medium (GM) and carob extract medium (CEM) by using recombinant Aspergillus sojae. The highest β-mannanase activities were obtained from microparticle-enhanced bioreactor strategy. It was found to be 347.47 U/mL by adding 10 g/L of Al 2 O 3 to GM and 439.13 U/mL by adding 1 g/L of talcum into CEM. The maximum β-mannanase activities for suspended, immobilization, and biofilm reactor remained at 72.55 U/mL in GM, 148.81 U/mL in CEM, and 194.09 U/mL in GM, respectively. The reason for that is the excessive, and irregular shaped growth and bulk formation, inadequate oxygen transfer or substrate diffusion in bioreactor. Consequently, the enzyme activity was significantly enhanced by addition of microparticles compared to other fed-batch fermentation strategies. Also, repeatable β-mannanase activities were obtained by controlling of the cell morphology by adding microparticle inside the fermenter.

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.

Auto-tuning system for NMR probe with LabView

NASA Astrophysics Data System (ADS)

Quen, Carmen; Mateo, Olivia; Bernal, Oscar

2013-03-01

Typical manual NMR-tuning method is not suitable for broadband spectra spanning several megahertz linewidths. Among the main problems encountered during manual tuning are pulse-power reproducibility, baselines, and transmission line reflections, to name a few. We present a design of an auto-tuning system using graphic programming language, LabVIEW, to minimize these problems. The program is designed to analyze the detected power signal of an antenna near the NMR probe and use this analysis to automatically tune the sample coil to match the impedance of the spectrometer (50 Ω). The tuning capacitors of the probe are controlled by a stepper motor through a LabVIEW/computer interface. Our program calculates the area of the power signal as an indicator to control the motor so disconnecting the coil to tune it through a network analyzer is unnecessary. Work supported by NSF-DMR 1105380

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.

[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.

Compatibility of refractory materials for nuclear reactor poison control systems

NASA Technical Reports Server (NTRS)

Sinclair, J. H.

1974-01-01

Metal-clad poison rods have been considered for the control system of an advanced space power reactor concept studied at the NASA Lewis Research Center. Such control rods may be required to operate at temperatures of about 140O C. Selected poison materials (including boron carbide and the diborides of zirconium, hafnium, and tantalum) were subjected to 1000-hour screening tests in contact with candidate refractory metal cladding materials (including tungsten and alloys of tantalum, niobium, and molybdenum) to assess the compatibility of these materials combinations at the temperatures of interest. Zirconium and hafnium diborides were compatible with refractory metals at 1400 C, but boron carbide and tantalum diboride reacted with the refractory metals at this temperature. Zirconium diboride also showed promise as a reaction barrier between boron carbide and tungsten.

Comparison of online reporting systems and their compatibility check with respective adverse drug reaction reporting forms

PubMed Central

Maharshi, Vikas; Nagar, Pravesh

2017-01-01

AIM: Different forms and online tools are available in different countries for spontaneous reporting, one of the most widely used methods of pharmacovigilance. Capturing sufficient information and adequate compatibility of online systems with respective reporting form is highly desirable for appropriate reporting of adverse drug reactions (ADRs). This study was aimed to compare three major online reporting systems (US, UK, and WHO) of the world and also to check their compatibility with the respective ADR reporting form. MATERIALS AND METHODS: A total of 89 data elements to provide relevant information were found out from above three online reporting systems. All three online systems were compared regarding magnitude of information captured by each of them and scoring was done by providing a score of “1” to each element. Compatibility of ADR reporting forms of India (Red form), US (Form 3500), and UK (Yellow card form) was assessed by comparing the information gathered by them with that can be entered into their respective online reporting systems, namely, “VigiFlow,” “US online reporting,” and “Yellow card online reporting.” Each unmatching item was given a score of “−1”. RESULTS: VigiFlow scored “74” points, whereas online reporting systems of the US and UK scored “56” and “49,” respectively, regarding magnitude of the information gathered by them. Compatibility score was found to be “0,” “−9,” and “−26” in case of ADR reporting systems of US, UK, and India, respectively. CONCLUSION: Our study reveals that “VigiFlow” is capable of capturing the maximum amount of information but “Form 3500” and “Online reporting system of US” are maximally compatible to each other among ADR reporting systems of all three countries. PMID:29515278

SPINS: standardized protein NMR storage. A data dictionary and object-oriented relational database for archiving protein NMR spectra.

PubMed

Baran, Michael C; Moseley, Hunter N B; Sahota, Gurmukh; Montelione, Gaetano T

2002-10-01

Modern protein NMR spectroscopy laboratories have a rapidly growing need for an easily queried local archival system of raw experimental NMR datasets. SPINS (Standardized ProteIn Nmr Storage) is an object-oriented relational database that provides facilities for high-volume NMR data archival, organization of analyses, and dissemination of results to the public domain by automatic preparation of the header files required for submission of data to the BioMagResBank (BMRB). The current version of SPINS coordinates the process from data collection to BMRB deposition of raw NMR data by standardizing and integrating the storage and retrieval of these data in a local laboratory file system. Additional facilities include a data mining query tool, graphical database administration tools, and a NMRStar v2. 1.1 file generator. SPINS also includes a user-friendly internet-based graphical user interface, which is optionally integrated with Varian VNMR NMR data collection software. This paper provides an overview of the data model underlying the SPINS database system, a description of its implementation in Oracle, and an outline of future plans for the SPINS project.

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.

Modeling of leachate recirculation using combined drainage blanket-horizontal trench systems in bioreactor landfills.

PubMed

Feng, Shi-Jin; Cao, Ben-Yi; Xie, Hai-Jian

2017-10-01

Leachate recirculation in municipal solid waste (MSW) landfills operated as bioreactors offers significant economic and environmental benefits. Combined drainage blanket (DB)-horizontal trench (HT) systems can be an alternative to single conventional recirculation approaches and can have competitive advantages. The key objectives of this study are to investigate combined drainage blanket -horizontal trench systems, to analyze the effects of applying two recirculation systems on the leachate migration in landfills, and to estimate some key design parameters (e.g., the steady-state flow rate, the influence width, and the cumulative leachate volume). It was determined that an effective recirculation model should consist of a moderate horizontal trench injection pressure head and supplementary leachate recirculated through drainage blanket, with an objective of increasing the horizontal unsaturated hydraulic conductivity and thereby allowing more leachate to flow from the horizontal trench system in a horizontal direction. In addition, design charts for engineering application were established using a dimensionless variable formulation.

Bioreactor validation and biocompatibility of Ag/poly(N-vinyl-2-pyrrolidone) hydrogel nanocomposites.

PubMed

Jovanović, Zeljka; Radosavljević, Aleksandra; Kačarević-Popović, Zorica; Stojkovska, Jasmina; Perić-Grujić, Aleksandra; Ristić, Mirjana; Matić, Ivana Z; Juranić, Zorica D; Obradovic, Bojana; Mišković-Stanković, Vesna

2013-05-01

Silver/poly(N-vinyl-2-pyrrolidone) (Ag/PVP) nanocomposites containing Ag nanoparticles at different concentrations were synthesized using γ-irradiation. Cytotoxicity of the obtained nanocomposites was determined by MTT assay in monolayer cultures of normal human immunocompetent peripheral blood mononuclear cells (PBMC) that were either non-stimulated or stimulated to proliferate by mitogen phytohemagglutinin (PHA), as well as in human cervix adenocarcinoma cell (HeLa) cultures. Silver release kinetics and mechanical properties of nanocomposites were investigated under bioreactor conditions in the simulated body fluid (SBF) at 37°C. The release of silver was monitored under static conditions, and in two types of bioreactors: perfusion bioreactors and a bioreactor with dynamic compression coupled with SBF perfusion simulating in vivo conditions in articular cartilage. Ag/PVP nanocomposites exhibited slight cytotoxic effects against PBMC at the estimated concentration of 0.4 μmol dm(-3), with negligible variations observed amongst different cell cultures investigated. Studies of the silver release kinetics indicated internal diffusion as the rate limiting step, determined by statistically comparable results obtained at all investigated conditions. However, silver release rate was slightly higher in the bioreactor with dynamic compression coupled with SBF perfusion as compared to the other two systems indicating the influence of dynamic compression. Modelling of silver release kinetics revealed potentials for optimization of Ag/PVP nanocomposites for particular applications as wound dressings or soft tissue implants. Copyright © 2013 Elsevier B.V. All rights reserved.

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

Cell culture experiments planned for the space bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R.; Cross, John H.

1987-01-01

Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.

Application of enhanced membrane bioreactor (eMBR) to treat dye wastewater.

PubMed

Rondon, Hector; El-Cheikh, William; Boluarte, Ida Alicia Rodriguez; Chang, Chia-Yuan; Bagshaw, Steve; Farago, Leanne; Jegatheesan, Veeriah; Shu, Li

2015-05-01

An enhanced membrane bioreactor (eMBR) consisting of two anoxic bioreactors (ARs) followed by an aerated membrane bioreactor (AMBR), UV-unit and a granular activated carbon (GAC) filter was employed to treat 50-100 mg/L of remazol blue BR dye. The COD of the feed was 2334 mg/L and COD:TN:TP in the feed was 119:1.87:1. A feed flow rate of 5 L/d was maintained when the dye concentration was 50 mg/L; 10 L/d of return activated sludge was recirculated to each AR from the AMBR. Once the biological system is acclimatised, 95% of dye, 99% of COD, 97% of nitrogen and 73% of phosphorus were removed at a retention time of 74.4 h. When the effluent from the AMBR was drawn at a flux rate of 6.5 L/m(2)h, the trans-membrane pressure reached 40 kPa in every 10 days. AMBR effluent was passed through the UV-unit and GAC filter to remove the dye completely. Copyright © 2015 Elsevier Ltd. All rights reserved.

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.

Correlation between mass transfer coefficient kLa and relevant operating parameters in cylindrical disposable shaken bioreactors on a bench-to-pilot scale

PubMed Central

2013-01-01

Background Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen kLa and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal kLa correlation. Results A Respiration Activity Monitoring System (RAMOS) was used to measure kLa values in cylindrical disposable shaken bioreactors and Buckingham’s π-Theorem was applied to define a dimensionless equation for kLa. In this way, a scale- and volume-independent kLa correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures. Conclusion The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/− 30%. To our knowledge, this is the first kLa correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale. PMID:24289110

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...

Hydrodynamic characteristics and overall volumetric oxygen transfer coefficient of a new multi-environment bioreactor.

PubMed

Behzadian, Farnaz; Yerushalmi, Laleh; Alimahmoodi, Mahmood; Mulligan, Catherine N

2013-08-01

The hydrodynamic characteristics and the overall volumetric oxygen transfer coefficient of a new multi-environment bioreactor which is an integrated part of a wastewater treatment system, called BioCAST, were studied. This bioreactor contains several zones with different environmental conditions including aerobic, microaerophilic and anoxic, designed to increase the contaminant removal capacity of the treatment system. The multi-environment bioreactor is designed based on the concept of airlift reactors where liquid is circulated through the zones with different environmental conditions. The presence of openings between the aerobic zone and the adjacent oxygen-depleted microaerophilic zone changes the hydrodynamic properties of this bioreactor compared to the conventional airlift designs. The impact of operating and process parameters, notably the hydraulic retention time (HRT) and superficial gas velocity (U(G)), on the hydrodynamics and mass transfer characteristics of the system was examined. The results showed that liquid circulation velocity (V(L)), gas holdup (ε) and overall volumetric oxygen transfer coefficient (k(L)a(L)) increase with the increase of superficial gas velocity (U(G)), while the mean circulation time (t(c)) decreases with the increase of superficial gas velocity. The mean circulation time between the aerobic zone (riser) and microaerophilic zone (downcomer) is a stronger function of the superficial gas velocity for the smaller openings (1/2 in.) between the two zones, while for the larger opening (1 in.) the mean circulation time is almost independent of U(G) for U(G) ≥ 0.023 m/s. The smaller openings between the two zones provide higher mass transfer coefficient and better zone generation which will contribute to improved performance of the system during treatment operations.

Magnetic resonance imaging compatible remote catheter navigation system with 3 degrees of freedom.

PubMed

Tavallaei, M A; Lavdas, M K; Gelman, D; Drangova, M

2016-08-01

To facilitate MRI-guided catheterization procedures, we present an MRI-compatible remote catheter navigation system that allows remote navigation of steerable catheters with 3 degrees of freedom. The system consists of a user interface (master), a robot (slave), and an ultrasonic motor control servomechanism. The interventionalist applies conventional motions (axial, radial and plunger manipulations) on an input catheter in the master unit; this user input is measured and used by the servomechanism to control a compact catheter manipulating robot, such that it replicates the interventionalist's input motion on the patient catheter. The performance of the system was evaluated in terms of MRI compatibility (SNR and artifact), feasibility of remote navigation under real-time MRI guidance, and motion replication accuracy. Real-time MRI experiments demonstrated that catheter was successfully navigated remotely to desired target references in all 3 degrees of freedom. The system had an absolute value error of [Formula: see text]1 mm in axial catheter motion replication over 30 mm of travel and [Formula: see text] for radial catheter motion replication over [Formula: see text]. The worst case SNR drop was observed to be [Formula: see text]3 %; the robot did not introduce any artifacts in the MR images. An MRI-compatible compact remote catheter navigation system has been developed that allows remote navigation of steerable catheters with 3 degrees of freedom. The proposed system allows for safe and accurate remote catheter navigation, within conventional closed-bore scanners, without degrading MR image quality.

UNITED STATES DEPARTMENT OF TRANSPORTATION GLOBAL POSITIONING SYSTEM (GPS) ADJACENT BAND COMPATIBILITY ASSESSMENT

DOT National Transportation Integrated Search

2018-04-01

The goal of the U.S. Department of Transportation (DOT) Global Positioning System (GPS) Adjacent Band Compatibility Assessment is to evaluate the maximum transmitted power levels of adjacent band radiofrequency (RF) systems that can be tolerated by G...

Bioreactor perfusion system for the long-term maintenance of tissue-engineered skeletal muscle organoids

NASA Technical Reports Server (NTRS)

Chromiak, J. A.; Shansky, J.; Perrone, C.; Vandenburgh, H. H.

1998-01-01

Three-dimensional skeletal muscle organ-like structures (organoids) formed in tissue culture by fusion of proliferating myoblasts into parallel networks of long, unbranched myofibers provide an in vivo-like model for examining the effects of growth factors, tension, and space flight on muscle cell growth and metabolism. To determine the feasibility of maintaining either avian or mammalian muscle organoids in a commercial perfusion bioreactor system, we measured metabolism, protein turnover. and autocrine/paracrine growth factor release rates. Medium glucose was metabolized at a constant rate in both low-serum- and serum-free media for up to 30 d. Total organoid noncollagenous protein and DNA content decreased approximately 22-28% (P < 0.05) over a 13-d period. Total protein synthesis rates could be determined accurately in the bioreactors for up to 30 h and total protein degradation rates could be measured for up to 3 wk. Special fixation and storage conditions necessary for space flight studies were validated as part of the studies. For example, the anabolic autocrine/paracrine skeletal muscle growth factors prostaglandin F2alpha (PGF2alpha) and insulin-like growth factor-1 (IGF-1) could be measured accurately in collected media fractions, even after storage at 37 degrees C for up to 10 d. In contrast, creatine kinase activity (a marker of cell damage) in collected media fractions was unreliable. These results provide initial benchmarks for long-term ex vivo studies of tissue-engineered skeletal muscle.

Production of transgenic strawberries by temporary immersion bioreactor system and verification by TAIL-PCR

PubMed Central

Hanhineva, Kati J; Kärenlampi, Sirpa O

2007-01-01

Background Strawberry (Fragaria × ananassa) is an economically important soft fruit crop with polyploid genome which complicates the breeding of new cultivars. For certain traits, genetic engineering offers a potential alternative to traditional breeding. However, many strawberry varieties are quite recalcitrant for Agrobacterium-mediated transformation, and a method allowing easy handling of large amounts of starting material is needed. Also the genotyping of putative transformants is challenging since the isolation of DNA for Southern analysis is difficult due to the high amount of phenolic compounds and polysaccharides that complicate efficient extraction of digestable DNA. There is thus a need to apply a screening method that is sensitive and unambiguous in identifying the different transformation events. Results Hygromycin-resistant strawberries were developed in temporary immersion bioreactors by Agrobacterium-mediated gene transfer. Putative transformants were screened by TAIL-PCR to verify T-DNA integration and to distinguish between the individual transformation events. Several different types of border sequence arrangements were detected. Conclusion This study demonstrates that temporary immersion bioreactor system suits well for the regeneration of transgenic strawberry plants as a labour-efficient technique. Small amount of DNA required by TAIL-PCR is easily recovered even from a small transformant, which allows rapid verification of T-DNA integration and detection of separate gene transfer events. These techniques combined clearly facilitate the generation of transgenic strawberries but should be applicable to other plants as well. PMID:17309794

jsNMR: an embedded platform-independent NMR spectrum viewer.

PubMed

Vosegaard, Thomas

2015-04-01

jsNMR is a lightweight NMR spectrum viewer written in JavaScript/HyperText Markup Language (HTML), which provides a cross-platform spectrum visualizer that runs on all computer architectures including mobile devices. Experimental (and simulated) datasets are easily opened in jsNMR by (i) drag and drop on a jsNMR browser window, (ii) by preparing a jsNMR file from the jsNMR web site, or (iii) by mailing the raw data to the jsNMR web portal. jsNMR embeds the original data in the HTML file, so a jsNMR file is a self-transforming dataset that may be exported to various formats, e.g. comma-separated values. The main applications of jsNMR are to provide easy access to NMR data without the need for dedicated software installed and to provide the possibility to visualize NMR spectra on web sites. Copyright © 2015 John Wiley & Sons, Ltd.

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

Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor

PubMed Central

Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, JH David; Bottaro, Andrea

2011-01-01

Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g. T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. PMID:21309085

Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor.

PubMed

Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, J H David; Bottaro, Andrea

2011-06-01

Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g., T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological, and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. Copyright © 2011 Wiley Periodicals, Inc.

GMP-Compliant Expansion of Clinical-Grade Human Mesenchymal Stromal/Stem Cells Using a Closed Hollow Fiber Bioreactor.

PubMed

Barckhausen, Christina; Rice, Brent; Baila, Stefano; Sensebé, Luc; Schrezenmeier, Hubert; Nold, Philipp; Hackstein, Holger; Rojewski, Markus Thomas

2016-01-01

This chapter describes a method for GMP-compliant expansion of human mesenchymal stromal/stem cells (hMSC) from bone marrow aspirates, using the Quantum(®) Cell Expansion System from Terumo BCT. The Quantum system is a functionally closed, automated hollow fiber bioreactor system designed to reproducibly grow cells in either GMP or research laboratory environments. The chapter includes protocols for preparation of media, setup of the Quantum system, coating of the hollow fiber bioreactor, as well as loading, feeding, and harvesting of cells. We suggest a panel of quality controls for the starting material, the interim product, as well as the final product.

Distributed and Lumped Parameter Models for the Characterization of High Throughput Bioreactors

PubMed Central

Conoscenti, Gioacchino; Cutrì, Elena; Tuan, Rocky S.; Raimondi, Manuela T.; Gottardi, Riccardo

2016-01-01

Next generation bioreactors are being developed to generate multiple human cell-based tissue analogs within the same fluidic system, to better recapitulate the complexity and interconnection of human physiology [1, 2]. The effective development of these devices requires a solid understanding of their interconnected fluidics, to predict the transport of nutrients and waste through the constructs and improve the design accordingly. In this work, we focus on a specific model of bioreactor, with multiple input/outputs, aimed at generating osteochondral constructs, i.e., a biphasic construct in which one side is cartilaginous in nature, while the other is osseous. We next develop a general computational approach to model the microfluidics of a multi-chamber, interconnected system that may be applied to human-on-chip devices. This objective requires overcoming several challenges at the level of computational modeling. The main one consists of addressing the multi-physics nature of the problem that combines free flow in channels with hindered flow in porous media. Fluid dynamics is also coupled with advection-diffusion-reaction equations that model the transport of biomolecules throughout the system and their interaction with living tissues and C constructs. Ultimately, we aim at providing a predictive approach useful for the general organ-on-chip community. To this end, we have developed a lumped parameter approach that allows us to analyze the behavior of multi-unit bioreactor systems with modest computational effort, provided that the behavior of a single unit can be fully characterized. PMID:27669413

Distributed and Lumped Parameter Models for the Characterization of High Throughput Bioreactors.

PubMed

Iannetti, Laura; D'Urso, Giovanna; Conoscenti, Gioacchino; Cutrì, Elena; Tuan, Rocky S; Raimondi, Manuela T; Gottardi, Riccardo; Zunino, Paolo

Next generation bioreactors are being developed to generate multiple human cell-based tissue analogs within the same fluidic system, to better recapitulate the complexity and interconnection of human physiology [1, 2]. The effective development of these devices requires a solid understanding of their interconnected fluidics, to predict the transport of nutrients and waste through the constructs and improve the design accordingly. In this work, we focus on a specific model of bioreactor, with multiple input/outputs, aimed at generating osteochondral constructs, i.e., a biphasic construct in which one side is cartilaginous in nature, while the other is osseous. We next develop a general computational approach to model the microfluidics of a multi-chamber, interconnected system that may be applied to human-on-chip devices. This objective requires overcoming several challenges at the level of computational modeling. The main one consists of addressing the multi-physics nature of the problem that combines free flow in channels with hindered flow in porous media. Fluid dynamics is also coupled with advection-diffusion-reaction equations that model the transport of biomolecules throughout the system and their interaction with living tissues and C constructs. Ultimately, we aim at providing a predictive approach useful for the general organ-on-chip community. To this end, we have developed a lumped parameter approach that allows us to analyze the behavior of multi-unit bioreactor systems with modest computational effort, provided that the behavior of a single unit can be fully characterized.

NMReDATA, a standard to report the NMR assignment and parameters of organic compounds.

PubMed

Pupier, Marion; Nuzillard, Jean-Marc; Wist, Julien; Schlörer, Nils E; Kuhn, Stefan; Erdelyi, Mate; Steinbeck, Christoph; Williams, Antony J; Butts, Craig; Claridge, Tim D W; Mikhova, Bozhana; Robien, Wolfgang; Dashti, Hesam; Eghbalnia, Hamid R; Farès, Christophe; Adam, Christian; Kessler, Pavel; Moriaud, Fabrice; Elyashberg, Mikhail; Argyropoulos, Dimitris; Pérez, Manuel; Giraudeau, Patrick; Gil, Roberto R; Trevorrow, Paul; Jeannerat, Damien

2018-04-14

Even though NMR has found countless applications in the field of small molecule characterization, there is no standard file format available for the NMR data relevant to structure characterization of small molecules. A new format is therefore introduced to associate the NMR parameters extracted from 1D and 2D spectra of organic compounds to the proposed chemical structure. These NMR parameters, which we shall call NMReDATA (for nuclear magnetic resonance extracted data), include chemical shift values, signal integrals, intensities, multiplicities, scalar coupling constants, lists of 2D correlations, relaxation times, and diffusion rates. The file format is an extension of the existing Structure Data Format, which is compatible with the commonly used MOL format. The association of an NMReDATA file with the raw and spectral data from which it originates constitutes an NMR record. This format is easily readable by humans and computers and provides a simple and efficient way for disseminating results of structural chemistry investigations, allowing automatic verification of published results, and for assisting the constitution of highly needed open-source structural databases. Copyright © 2018 John Wiley & Sons, Ltd.

Mathematical modeling of the integrated process of mercury bioremediation in the industrial bioreactor.

PubMed

Głuszcz, Paweł; Petera, Jerzy; Ledakowicz, Stanisław

2011-03-01

The mathematical model of the integrated process of mercury contaminated wastewater bioremediation in a fixed-bed industrial bioreactor is presented. An activated carbon packing in the bioreactor plays the role of an adsorbent for ionic mercury and at the same time of a carrier material for immobilization of mercury-reducing bacteria. The model includes three basic stages of the bioremediation process: mass transfer in the liquid phase, adsorption of mercury onto activated carbon and ionic mercury bioreduction to Hg(0) by immobilized microorganisms. Model calculations were verified using experimental data obtained during the process of industrial wastewater bioremediation in the bioreactor of 1 m³ volume. It was found that the presented model reflects the properties of the real system quite well. Numerical simulation of the bioremediation process confirmed the experimentally observed positive effect of the integration of ionic mercury adsorption and bioreduction in one apparatus.

Hepatic Differentiation and Maturation of Human Embryonic Stem Cells Cultured in a Perfused Three-Dimensional Bioreactor

PubMed Central

Synnergren, Jane; Jensen, Janne; Björquist, Petter; Ingelman-Sundberg, Magnus

2013-01-01

Drug-induced liver injury is a serious and frequently occurring adverse drug reaction in the clinics and is hard to predict during preclinical studies. Today, primary hepatocytes are the most frequently used cell model for drug discovery and prediction of toxicity. However, their use is marred by high donor variability regarding drug metabolism and toxicity, and instable expression levels of liver-specific genes such as cytochromes P450. An in vitro model system based on human embryonic stem cells (hESC), with their unique properties of pluripotency and self-renewal, has potential to provide a stable and unlimited supply of human hepatocytes. Much effort has been made to direct hESC toward the hepatic lineage, mostly using 2-dimensional (2D) cultures. Although the results are encouraging, these cells lack important functionality. Here, we investigate if hepatic differentiation of hESC can be improved by using a 3-dimensional (3D) bioreactor system. Human ESCs were differentiated toward the hepatic lineage using the same cells in either the 3D or 2D system. A global transcriptional analysis identified important differences between the 2 differentiation regimes, and we identified 10 pathways, highly related to liver functions, which were significantly upregulated in cells differentiated in the bioreactor compared to 2D control cultures. The enhanced hepatic differentiation observed in the bioreactor system was also supported by immunocytochemistry. Taken together, our results suggest that hepatic differentiation of hESC is improved when using this 3D bioreactor technology as compared to 2D culture systems. PMID:22970843

Hepatic differentiation and maturation of human embryonic stem cells cultured in a perfused three-dimensional bioreactor.

PubMed

Sivertsson, Louise; Synnergren, Jane; Jensen, Janne; Björquist, Petter; Ingelman-Sundberg, Magnus

2013-02-15

Drug-induced liver injury is a serious and frequently occurring adverse drug reaction in the clinics and is hard to predict during preclinical studies. Today, primary hepatocytes are the most frequently used cell model for drug discovery and prediction of toxicity. However, their use is marred by high donor variability regarding drug metabolism and toxicity, and instable expression levels of liver-specific genes such as cytochromes P450. An in vitro model system based on human embryonic stem cells (hESC), with their unique properties of pluripotency and self-renewal, has potential to provide a stable and unlimited supply of human hepatocytes. Much effort has been made to direct hESC toward the hepatic lineage, mostly using 2-dimensional (2D) cultures. Although the results are encouraging, these cells lack important functionality. Here, we investigate if hepatic differentiation of hESC can be improved by using a 3-dimensional (3D) bioreactor system. Human ESCs were differentiated toward the hepatic lineage using the same cells in either the 3D or 2D system. A global transcriptional analysis identified important differences between the 2 differentiation regimes, and we identified 10 pathways, highly related to liver functions, which were significantly upregulated in cells differentiated in the bioreactor compared to 2D control cultures. The enhanced hepatic differentiation observed in the bioreactor system was also supported by immunocytochemistry. Taken together, our results suggest that hepatic differentiation of hESC is improved when using this 3D bioreactor technology as compared to 2D culture systems.

Bioreactor Cultivation of Anatomically Shaped Human Bone Grafts

PubMed Central

Temple, Joshua P.; Yeager, Keith; Bhumiratana, Sarindr; Vunjak-Novakovic, Gordana; Grayson, Warren L.

2015-01-01

In this chapter, we describe a method for engineering bone grafts in vitro with the specific geometry of the temporomandibular joint (TMJ) condyle. The anatomical geometry of the bone grafts was segmented from computed tomography (CT) scans, converted to G-code, and used to machine decellularized trabecular bone scaffolds into the identical shape of the condyle. These scaffolds were seeded with human bone marrow-derived mesenchymal stem cells (MSCs) using spinner flasks and cultivated for up to 5 weeks in vitro using a custom-designed perfusion bioreactor system. The flow patterns through the complex geometry were modeled using the FloWorks module of SolidWorks to optimize bioreactor design. The perfused scaffolds exhibited significantly higher cellular content, better matrix production, and increased bone mineral deposition relative to non-perfused (static) controls after 5 weeks of in vitro cultivation. This technology is broadly applicable for creating patient-specific bone grafts of varying shapes and sizes. PMID:24014312

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.

Energy and greenhouse gas life cycle assessment and cost analysis of aerobic and anaerobic membrane bioreactor systems: Influence of scale, population density, climate, and methane recovery

EPA Science Inventory

This study calculated the energy and greenhouse gas life cycle and cost profiles of transitional aerobic membrane bioreactors (AeMBR) and anaerobic membrane bioreactors (AnMBR). Membrane bioreactors (MBR) represent a promising technology for decentralized wastewater treatment and...

Plastic carrier polishing chamber reduces pollution swapping from denitrifying woodchip bioreactors

USDA-ARS?s Scientific Manuscript database

Denitrifying bioreactors with solid organic carbon sources (i.e., “woodchip bioreactors”) have proven to be relatively simple and cost effective treatment systems for nitrate-laden agricultural and aquacultural waters and wastewaters. However, because this technology is still relatively new, design ...

Energy and greenhouse gas life cycle assessment and cost analysis of aerobic and anaerobic membrane bioreactor systems: Influence of scale, population density, climate, and methane recovery.

PubMed

Cashman, Sarah; Ma, Xin; Mosley, Janet; Garland, Jay; Crone, Brian; Xue, Xiaobo

2018-04-01

This study calculated the energy and greenhouse gas life cycle and cost profiles of transitional aerobic membrane bioreactors (AeMBR) and anaerobic membrane bioreactors (AnMBR). Membrane bioreactors (MBR) represent a promising technology for decentralized wastewater treatment and can produce recycled water to displace potable water. Energy recovery is possible with methane generated from AnMBRs. Scenarios for these technologies were investigated for different scale systems serving various population densities under a number of climate conditions with multiple methane recovery options. When incorporating the displacement of drinking water, AeMBRs started to realize net energy benefits at the 1 million gallons per day (MGD) scale and mesophilic AnMBRs at the 5 MGD scale. For all scales, the psychrophilic AnMBR resulted in net energy benefits. This study provides insights into key performance characteristics needed before an informed decision can be made for a community to transition towards the adoption of MBR technologies. Copyright © 2018. Published by Elsevier Ltd.

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.

Validation of Bioreactor and Human-on-a-Chip Devices for Chemical Safety Assessment.

PubMed

Rebelo, Sofia P; Dehne, Eva-Maria; Brito, Catarina; Horland, Reyk; Alves, Paula M; Marx, Uwe

2016-01-01

Equipment and device qualification and test assay validation in the field of tissue engineered human organs for substance assessment remain formidable tasks with only a few successful examples so far. The hurdles seem to increase with the growing complexity of the biological systems, emulated by the respective models. Controlled single tissue or organ culture in bioreactors improves the organ-specific functions and maintains their phenotypic stability for longer periods of time. The reproducibility attained with bioreactor operations is, per se, an advantage for the validation of safety assessment. Regulatory agencies have gradually altered the validation concept from exhaustive "product" to rigorous and detailed process characterization, valuing reproducibility as a standard for validation. "Human-on-a-chip" technologies applying micro-physiological systems to the in vitro combination of miniaturized human organ equivalents into functional human micro-organisms are nowadays thought to be the most elaborate solution created to date. They target the replacement of the current most complex models-laboratory animals. Therefore, we provide here a road map towards the validation of such "human-on-a-chip" models and qualification of their respective bioreactor and microchip equipment along a path currently used for the respective animal models.

NMR imaging of density distributions in tablets.

PubMed

Djemai, A; Sinka, I C

2006-08-17

This paper describes the use of (1)H nuclear magnetic resonance (NMR) for 3D mapping of the relative density distribution in pharmaceutical tablets manufactured under controlled conditions. The tablets are impregnated with a compatible liquid. The technique involves imaging of the presence of liquid which occupies the open pore space. The method does not require special calibration as the signal is directly proportional to the porosity for the imaging conditions used. The NMR imaging method is validated using uniform density flat faced tablets and also by direct comparison with X-ray computed tomography. The results illustrate (1) the effect of die wall friction on density distribution by compressing round, curved faced tablets using clean and pre-lubricated tooling, (2) the evolution of density distribution during compaction for both clean and pre-lubricated die wall conditions, by imaging tablets compressed to different compaction forces, and (3) the effect of tablet image on density distribution by compressing two complex shape tablets in identical dies to the same average density using punches with different geometries.

Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors

PubMed Central

van den Berg, Frans; Racher, Andrew J.; Martin, Elaine B.; Jaques, Colin

2017-01-01

Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in large‐scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D‐fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design‐of‐experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D‐fluorescence could more accurately measure ammonium concentration (RMSECV 0.031 g L−1) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSECV 1.11 and 0.92 g L−1, respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D‐fluorescence. The implementation of Raman spectroscopy increases at‐line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337–346, 2017 PMID:28271638

Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors.

PubMed

Rowland-Jones, Ruth C; van den Berg, Frans; Racher, Andrew J; Martin, Elaine B; Jaques, Colin

2017-03-01

Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in large-scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D-fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design-of-experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D-fluorescence could more accurately measure ammonium concentration (RMSE CV 0.031 g L -1 ) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSE CV 1.11 and 0.92 g L -1 , respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D-fluorescence. The implementation of Raman spectroscopy increases at-line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337-346, 2017. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on

Analytical Applications of NMR: Summer Symposium on Analytical Chemistry.

ERIC Educational Resources Information Center

Borman, Stuart A.

1982-01-01

Highlights a symposium on analytical applications of nuclear magnetic resonance spectroscopy (NMR), discussing pulse Fourier transformation technique, two-dimensional NMR, solid state NMR, and multinuclear NMR. Includes description of ORACLE, an NMR data processing system at Syracuse University using real-time color graphics, and algorithms for…

On Software Compatibility.

ERIC Educational Resources Information Center

Ershov, Andrei P.

The problem of compatibility of software hampers the development of computer application. One solution lies in standardization of languages, terms, peripherais, operating systems and computer characteristics. (AB)

Temperature and Substrate Control Woodchip Bioreactor Performance in Reducing Tile Nitrate Loads in East-Central Illinois.

PubMed

David, Mark B; Gentry, Lowell E; Cooke, Richard A; Herbstritt, Stephanie M

2016-05-01

Tile drainage is the major source of nitrate in the upper Midwest, and end-of-tile removal techniques such as wood chip bioreactors have been installed that allow current farming practices to continue, with nitrate removed through denitrification. There have been few multiyear studies of bioreactors examining controls on nitrate removal rates. We evaluated the nitrate removal performance of two wood chip bioreactors during the first 3 yr of operation and examined the major factors that regulated nitrate removal. Bioreactor 2 was subject to river flooding, and performance was not assessed. Bioreactor 1 had average monthly nitrate removal rates of 23 to 44 g N m d in Year 1, which decreased to 1.2 to 11 g N m d in Years 2 and 3. The greater N removal rates in Year 1 and early in Year 2 were likely due to highly degradable C in the woodchips. Only late in Year 2 and in Year 3 was there a strong temperature response in the nitrate removal rate. Less than 1% of the nitrate removed was emitted as NO. Due to large tile inputs of nitrate (729-2127 kg N) at high concentrations (∼30 mg nitrate N L) in Years 2 and 3, overall removal efficiency was low (3 and 7% in Years 2 and 3, respectively). Based on a process-based bioreactor performance model, Bioreactor 1 would have needed to be 9 times as large as the current system to remove 50% of the nitrate load from this 20-ha field. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Chip-based three-dimensional cell culture in perfused micro-bioreactors.

PubMed

Gottwald, Eric; Lahni, Brigitte; Thiele, David; Giselbrecht, Stefan; Welle, Alexander; Weibezahn, Karl-Friedrich

2008-05-21

We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from non-biodegradable polymers, e.g., polycarbonate or polymethyl methacrylate by micro injection molding, micro hot embossing or micro thermo-forming. But, it can also be manufactured from bio-degradable polymers. Its overall dimensions are 0.7 1 x 20 x 20 x 0.7 1 mm (h x w x l). The main features of the chips used are either a grid of up to 1156 cubic micro-containers (cf-chip) each the size of 120-300 x 300 x 300 micron (h x w x l) or round recesses with diameters of 300 micron and a depth of 300 micron (r-chip). The scaffold can house 10 Mio. cells in a three-dimensional configuration. For an optimal nutrient and gas supply, the chip is inserted in a bioreactor housing. The bioreactor is part of a closed sterile circulation loop that, in the simplest configuration, is additionally comprised of a roller pump and a medium reservoir with a gas supply. The bioreactor can be run in perfusion, superfusion, or even a mixed operation mode. We have successfully cultivated cell lines as well as primary cells over periods of several weeks. For rat primary liver cells we could show a preservation of organotypic functions for more than 2 weeks. For hepatocellular carcinoma cell lines we could show the induction of liver specific genes not or only slightly expressed in standard monolayer culture. The system might also be useful as a stem cell cultivation system since first differentiation experiments with stem cell lines were promising.

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.

Automated sequence-specific protein NMR assignment using the memetic algorithm MATCH.

PubMed

Volk, Jochen; Herrmann, Torsten; Wüthrich, Kurt

2008-07-01

MATCH (Memetic Algorithm and Combinatorial Optimization Heuristics) is a new memetic algorithm for automated sequence-specific polypeptide backbone NMR assignment of proteins. MATCH employs local optimization for tracing partial sequence-specific assignments within a global, population-based search environment, where the simultaneous application of local and global optimization heuristics guarantees high efficiency and robustness. MATCH thus makes combined use of the two predominant concepts in use for automated NMR assignment of proteins. Dynamic transition and inherent mutation are new techniques that enable automatic adaptation to variable quality of the experimental input data. The concept of dynamic transition is incorporated in all major building blocks of the algorithm, where it enables switching between local and global optimization heuristics at any time during the assignment process. Inherent mutation restricts the intrinsically required randomness of the evolutionary algorithm to those regions of the conformation space that are compatible with the experimental input data. Using intact and artificially deteriorated APSY-NMR input data of proteins, MATCH performed sequence-specific resonance assignment with high efficiency and robustness.

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

Dynamic Bioreactor Culture of High Volume Engineered Bone Tissue

PubMed Central

Nguyen, Bao-Ngoc B.; Ko, Henry; Moriarty, Rebecca A.; Etheridge, Julie M.

2016-01-01

Within the field of tissue engineering and regenerative medicine, the fabrication of tissue grafts of any significant size—much less a whole organ or tissue—remains a major challenge. Currently, tissue-engineered constructs cultured in vitro have been restrained in size primarily due to the diffusion limit of oxygen and nutrients to the center of these grafts. Previously, we developed a novel tubular perfusion system (TPS) bioreactor, which allows the dynamic culture of bead-encapsulated cells and increases the supply of nutrients to the entire cell population. More interestingly, the versatility of TPS bioreactor allows a large range of engineered tissue volumes to be cultured, including large bone grafts. In this study, we utilized alginate-encapsulated human mesenchymal stem cells for the culture of a tissue-engineered bone construct in the size and shape of the superior half of an adult human femur (∼200 cm3), a 20-fold increase over previously reported volumes of in vitro engineered bone grafts. Dynamic culture in TPS bioreactor not only resulted in high cell viability throughout the femur graft, but also showed early signs of stem cell differentiation through increased expression of osteogenic genes and proteins, consistent with our previous models of smaller bone constructs. This first foray into full-scale bone engineering provides the foundation for future clinical applications of bioengineered bone grafts. PMID:26653703

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.

Design and validation of a pulsatile perfusion bioreactor for 3D high cell density cultures.

PubMed

Chouinard, Julie A; Gagnon, Serge; Couture, Marc G; Lévesque, Alain; Vermette, Patrick

2009-12-15

This study presents the design and validation of a pulsatile flow perfusion bioreactor able to provide a suitable environment for 3D high cell density cultures for tissue engineering applications. Our bioreactor system is mobile, does not require the use of traditional cell culture incubators and is easy to sterilize. It provides real-time monitoring and stable control of pH, dissolved oxygen concentration, temperature, pressure, pulsation frequency, and flow rate. In this bioreactor system, cells are cultured in a gel within a chamber perfused by a culture medium fed by hollow fibers. Human umbilical vein endothelial cells (HUVEC) suspended in fibrin were found to be living, making connections and proliferating up to five to six times their initial seeding number after a 48-h culture period. Cells were uniformly dispersed within the 14.40 mm x 17.46 mm x 6.35 mm chamber. A larger fraction of the cells suspended in 6.35-mm thick gels and cultured in a traditional CO(2) incubator were found to be round and dead [corrected]. In control experiments carried out in a traditional cell culture incubator, the scarcely found living cells were mostly on top of the gels, while cells cultured under perfusion bioreactor conditions were found to be alive and uniformly distributed across the gel. 2009 Wiley Periodicals, Inc.

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.

Oxygen Compatibility Assessment of Components and Systems

NASA Technical Reports Server (NTRS)

Stoltzfus, Joel; Sparks, Kyle

2010-01-01

Fire hazards are inherent in oxygen systems and a storied history of fires in rocket engine propulsion components exists. To detect and mitigate these fire hazards requires careful, detailed, and thorough analyses applied during the design process. The oxygen compatibility assessment (OCA) process designed by NASA Johnson Space Center (JSC) White Sands Test Facility (WSTF) can be used to determine the presence of fire hazards in oxygen systems and the likelihood of a fire. This process may be used as both a design guide and during the approval process to ensure proper design features and material selection. The procedure for performing an OCA is a structured step-by-step process to determine the most severe operating conditions; assess the flammability of the system materials at the use conditions; evaluate the presence and efficacy of ignition mechanisms; assess the potential for a fire to breach the system; and determine the reaction effect (the potential loss of life, mission, and system functionality as the result of a fire). This process should be performed for each component in a system. The results of each component assessment, and the overall system assessment, should be recorded in a report that can be used in the short term to communicate hazards and their mitigation and to aid in system/component development and, in the long term, to solve anomalies that occur during engine testing and operation.

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

Treatment of coal gasification wastewater by membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system.

PubMed

Jia, Shengyong; Han, Hongjun; Hou, Baolin; Zhuang, Haifeng; Fang, Fang; Zhao, Qian

2014-12-01

A laboratory-scale membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system was developed to treat coal gasification wastewater to enhance the COD, total phenols (TPh), NH4+ removals and migrate the membrane fouling. Since the MBR–PAC system operated with PAC dosage of 4 g L−1, the maximum removal efficiencies of COD, TPh and NH4+ reached 93%, 99% and 63%, respectively with the corresponding influent concentrations of 2.27 g L−1, 497 mg L−1 and 164 mg N L−1; the PAC extraction efficiencies of COD, TPh and NH4+ were 6%, 3% and 13%, respectively; the transmembrane pressure decreased 34% with PAC after 50 d operation. The results demonstrate that PAC played a key role in the enhancement of biodegradability and mitigation of membrane fouling.

Bacterial community structure of a lab-scale anammox membrane bioreactor.

PubMed

Gonzalez-Martinez, Alejandro; Osorio, F; Rodriguez-Sanchez, Alejandro; Martinez-Toledo, Maria Victoria; Gonzalez-Lopez, Jesus; Lotti, Tommaso; van Loosdrecht, M C M

2015-01-01

Autotrophic nitrogen removal technologies have proliferated through the last decade. Among these, a promising one is the membrane bioreactor (MBR) Anammox, which can achieve very high solids retention time and therefore sets a proper environment for the cultivation of anammox bacteria. In this sense, the MBR Anammox is an efficient technology for the treatment of effluents with low organic carbon and high ammonium concentrations once it has been treated under partial nitrification systems. A lab-scale MBR Anammox bioreactor has been built at the Technological University of Delft, The Netherlands and has been proven for efficient nitrogen removal and efficient cultivation of anammox bacteria. In this study, next-generation sequencing techniques have been used for the investigation of the bacterial communities of this MBR Anammox for the first time ever. A strong domination of Candidatus Brocadia bacterium and also the presence of a myriad of other microorganisms that have adapted to this environment were detected, suggesting that the MBR Anammox bioreactor might have a more complex microbial ecosystem that it has been thought. Among these, nitrate-reducing heterotrophs and primary producers, among others, were identified. Definition of the ecological roles of the OTUs identified through metagenomic analysis was discussed. © 2014 American Institute of Chemical Engineers.

NMR characterization of thin films

DOEpatents

Gerald II, Rex E.; Klingler, Robert J.; Rathke, Jerome W.; Diaz, Rocio; Vukovic, Lela

2010-06-15

A method, apparatus, and system for characterizing thin film materials. The method, apparatus, and system includes a container for receiving a starting material, applying a gravitational force, a magnetic force, and an electric force or combinations thereof to at least the starting material, forming a thin film material, sensing an NMR signal from the thin film material and analyzing the NMR signal to characterize the thin film of material.

NMR characterization of thin films

DOEpatents

Gerald, II, Rex E.; Klingler, Robert J.; Rathke, Jerome W.; Diaz, Rocio; Vukovic, Lela

2008-11-25

A method, apparatus, and system for characterizing thin film materials. The method, apparatus, and system includes a container for receiving a starting material, applying a gravitational force, a magnetic force, and an electric force or combinations thereof to at least the starting material, forming a thin film material, sensing an NMR signal from the thin film material and analyzing the NMR signal to characterize the thin film of material.

A study of the Coriolis effect on the fluid flow profile in a centrifugal bioreactor.

PubMed

Detzel, Christopher J; Thorson, Michael R; Van Wie, Bernard J; Ivory, Cornelius F

2009-01-01

Increasing demand for tissues, proteins, and antibodies derived from cell culture is necessitating the development and implementation of high cell density bioreactors. A system for studying high density culture is the centrifugal bioreactor (CCBR), which retains cells by increasing settling velocities through system rotation, thereby eliminating diffusional limitations associated with mechanical cell retention devices. This article focuses on the fluid mechanics of the CCBR system by considering Coriolis effects. Such considerations for centrifugal bioprocessing have heretofore been ignored; therefore, a simpler analysis of an empty chamber will be performed. Comparisons are made between numerical simulations and bromophenol blue dye injection experiments. For the non-rotating bioreactor with an inlet velocity of 4.3 cm/s, both the numerical and experimental results show the formation of a teardrop shaped plume of dye following streamlines through the reactor. However, as the reactor is rotated, the simulation predicts the development of vortices and a flow profile dominated by Coriolis forces resulting in the majority of flow up the leading wall of the reactor as dye initially enters the chamber, results are confirmed by experimental observations. As the reactor continues to fill with dye, the simulation predicts dye movement up both walls while experimental observations show the reactor fills with dye from the exit to the inlet. Differences between the simulation and experimental observations can be explained by excessive diffusion required for simulation convergence, and a slight density difference between dyed and un-dyed solutions. Implications of the results on practical bioreactor use are also discussed. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

A Study of the Coriolis Effect on the Fluid Flow Profile in a Centrifugal Bioreactor

PubMed Central

Detzel, Christopher J.; Thorson, Michael R.; Van Wie, Bernard J.; Ivory, Cornelius F.

2011-01-01

Increasing demand for tissues, proteins, and antibodies derived from cell culture is necessitating the development and implementation of high cell density bioreactors. A system for studying high density culture is the centrifugal bioreactor (CCBR) which retains cells by increasing settling velocities through system rotation, thereby eliminating diffusional limitations associated with mechanical cell retention devices. This paper focuses on the fluid mechanics of the CCBR system by considering Coriolis effects. Such considerations for centrifugal bioprocessing have heretofore been ignored; therefore a simpler analysis of an empty chamber will be performed. Comparisons are made between numerical simulations and bromophenol blue dye injection experiments. For the non-rotating bioreactor with an inlet velocity of 4.3 cm/s, both the numerical and experimental results show the formation of a teardrop shaped plume of dye following streamlines through the reactor. However, as the reactor is rotated the simulation predicts the development of vortices and a flow profile dominated by Coriolis forces resulting in the majority of flow up the leading wall of the reactor as dye initially enters the chamber, results confirmed by experimental observations. As the reactor continues to fill with dye, the simulation predicts dye movement up both walls while experimental observations show the reactor fills with dye from the exit to the inlet. Differences between the simulation and experimental observations can be explained by excessive diffusion required for simulation convergence, and a slight density difference between dyed and un-dyed solutions. Implications of the results on practical bioreactor use are also discussed. PMID:19455639

Development of a Communication System Compatible with Chemical Protective Clothing and Equipment.

DTIC Science & Technology

1986-06-23

for the Chemical Protective Clothing Communication System are discussed in the operation manuals for the engineering prototypes of the trans- ceiver...Report Date DEVELOPMENT OF A COMMUNICATION SYSTEM COMPATIBLE June 1986 WITH CHEMICAL PROTECTIVE CLOTHING AND EQUIPMENT 6. Performing Organization Code 8...Abstract - ’The U. S. Coast Guard and NASA joined in a project to develop a cominications system to operate inside protective suits used in chemical spill

Phase separated membrane bioreactor: Results from model system studies

NASA Astrophysics Data System (ADS)

Petersen, G. R.; Seshan, P. K.; Dunlop, E. H.

The operation and evaluation of a bioreactor designed for high intensity oxygen transfer in a microgravity environment is described. The reactor itself consists of a zero headspace liquid phase separated from the air supply by a long length of silicone rubber tubing through which the oxygen diffuses in and the carbon dioxide diffuses out. Mass transfer studies show that the oxygen is film diffusion controlled both externally and internally to the tubing and not by diffusion across the tube walls. Methods of upgrading the design to eliminate these resistances are proposed. Cell growth was obtained in the fermenter using Saccharomyces cerevisiae showing that this concept is capable of sustaining cell growth in the terrestial simulation.

Phase separated membrane bioreactor - Results from model system studies

NASA Technical Reports Server (NTRS)

Petersen, G. R.; Seshan, P. K.; Dunlop, E. H.

1989-01-01

The operation and evaluation of a bioreactor designed for high intensity oxygen transfer in a microgravity environment is described. The reactor itself consists of a zero headspace liquid phase separated from the air supply by a long length of silicone rubber tubing through which the oxygen diffuses in and the carbon dioxide diffuses out. Mass transfer studies show that the oxygen is film diffusion controlled both externally and internally to the tubing and not by diffusion across the tube walls. Methods of upgrading the design to eliminate these resistances are proposed. Cell growth was obtained in the fermenter using Saccharomyces cerevisiae showing that this concept is capable of sustaining cell growth in the terrestrial simulation.

Model system studies with a phase separated membrane bioreactor

NASA Technical Reports Server (NTRS)

Petersen, G. R.; Seshan, P. K.; Dunlop, Eric H.

1989-01-01

The operation and evaluation of a bioreactor designed for high intensity oxygen transfer in a microgravity environment is described. The reactor itself consists of a zero headspace liquid phase separated from the air supply by a long length of silicone rubber tubing through which the oxygen diffuses in and the carbon dioxide diffuses out. Mass transfer studies show that the oxygen is film diffusion controlled both externally and internally to the tubing and not by diffusion across the tube walls. Methods of upgrading the design to eliminate these resistances are proposed. Cell growth was obtained in the fermenter using Saccharomyces cerevisiae showing that this concept is capable of sustaining cell growth in the terrestial simulation.

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

Ten years of industrial and municipal membrane bioreactor (MBR) systems - lessons from the field.

PubMed

Larrea, Asun; Rambor, Andre; Fabiyi, Malcolm

2014-01-01

The use of membrane bioreactors (MBRs) in activated sludge wastewater treatment has grown significantly in the last decade. While there is growing awareness and knowledge about the application of MBR technology in municipal wastewater treatment, not much information is available on the application of MBRs in industrial wastewater treatment. A comparative study of design data, operating conditions and the major challenges associated with MBR operations in 24 MBR plants treating both municipal and industrial wastewater, built by and/or operated by Praxair, Inc., is presented. Of the 24 MBR systems described, 12 of the plants used high purity oxygen (HPO). By enabling a wide range of food/microorganism ratios and loading conditions in the same system, HPO MBR systems can extend the options available to industrial plant operators to meet the challenges of wide fluctuations in organic loading and footprint limitations. While fouling in industrial MBR systems can be an issue, adequate flux and permeability values can be reliably maintained by the use of good maintenance strategies and effective process controls (pretreatment, cleaning and membrane autopsies).

A novel osmosis membrane bioreactor-membrane distillation hybrid system for wastewater treatment and reuse.

PubMed

Nguyen, Nguyen Cong; Nguyen, Hau Thi; Chen, Shiao-Shing; Ngo, Huu Hao; Guo, Wenshan; Chan, Wen Hao; Ray, Saikat Sinha; Li, Chi-Wang; Hsu, Hung-Te

2016-06-01

A novel approach was designed to simultaneously enhance nutrient removal and reduce membrane fouling for wastewater treatment using an attached growth biofilm (AGB) integrated with an osmosis membrane bioreactor (OsMBR) system for the first time. In this study, a highly charged organic compound (HEDTA(3-)) was employed as a novel draw solution in the AGB-OsMBR system to obtain a low reverse salt flux, maintain a healthy environment for the microorganisms. The AGB-OsMBR system achieved a stable water flux of 3.62L/m(2)h, high nutrient removal of 99% and less fouling during a 60-day operation. Furthermore, the high salinity of diluted draw solution could be effectively recovered by membrane distillation (MD) process with salt rejection of 99.7%. The diluted draw solution was re-concentrated to its initial status (56.1mS/cm) at recovery of 9.8% after 6h. The work demonstrated that novel multi-barrier systems could produce high quality potable water from impaired streams. Copyright © 2016. Published by Elsevier Ltd.

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

validated for primary and immortalized cells, and the design of each bioreactor is scalable to accommodate any three-dimensional scaffold (e.g., synthetic or naturally derived matrix) that contains conduits for nutrient perfusion to deliver media to growing cells and monitor noninvasive parameters during scaffold repopulation, broadening the applicability of these bioreactor systems. PMID:25929317

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

validated for primary and immortalized cells, and the design of each bioreactor is scalable to accommodate any three-dimensional scaffold (e.g., synthetic or naturally derived matrix) that contains conduits for nutrient perfusion to deliver media to growing cells and monitor noninvasive parameters during scaffold repopulation, broadening the applicability of these bioreactor systems.

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...

Solid-state NMR imaging system

DOEpatents

Gopalsami, Nachappa; Dieckman, Stephen L.; Ellingson, William A.

1992-01-01

An apparatus for use with a solid-state NMR spectrometer includes a special imaging probe with linear, high-field strength gradient fields and high-power broadband RF coils using a back projection method for data acquisition and image reconstruction, and a real-time pulse programmer adaptable for use by a conventional computer for complex high speed pulse sequences.

Development of a hybrid photo-bioreactor and nanoparticle adsorbent system for the removal of CO2, and selected organic and metal co-pollutants.

PubMed

Rocha, Andrea A; Wilde, Christian; Hu, Zhenzhong; Nepotchatykh, Oleg; Nazarenko, Yevgen; Ariya, Parisa A

2017-07-01

Fossil fuel combustion and many industrial processes generate gaseous emissions that contain a number of toxic organic pollutants and carbon dioxide (CO 2 ) which contribute to climate change and atmospheric pollution. There is a need for green and sustainable solutions to remove air pollutants, as opposed to conventional techniques which can be expensive, consume additional energy and generate further waste. We developed a novel integrated bioreactor combined with recyclable iron oxide nano/micro-particle adsorption interfaces, to remove CO 2, and undesired organic air pollutants using natural particles, while generating oxygen. This semi-continuous bench-scale photo-bioreactor was shown to successfully clean up simulated emission streams of up to 45% CO 2 with a conversion rate of approximately 4% CO 2 per hour, generating a steady supply of oxygen (6mmol/hr), while nanoparticles effectively remove several undesired organic by-products. We also showed algal waste of the bioreactor can be used for mercury remediation. We estimated the potential CO 2 emissions that could be captured from our new method for three industrial cases in which, coal, oil and natural gas were used. With a 30% carbon capture system, the reduction of CO 2 was estimated to decrease by about 420,000, 320,000 and 240,000 metric tonnes, respectively for a typical 500MW power plant. The cost analysis we conducted showed potential to scale-up, and the entire system is recyclable and sustainable. We further discuss the implications of usage of this complete system, or as individual units, that could provide a hybrid option to existing industrial setups. Copyright © 2016. Published by Elsevier B.V.

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.

Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies.

PubMed

Hoffmann, Stefan A; Müller-Vieira, Ursula; Biemel, Klaus; Knobeloch, Daniel; Heydel, Sandra; Lübberstedt, Marc; Nüssler, Andreas K; Andersson, Tommy B; Gerlach, Jörg C; Zeilinger, Katrin

2012-12-01

Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver. Copyright © 2012 Wiley Periodicals, Inc.

Achievement of a 920-MHz High Resolution NMR

NASA Astrophysics Data System (ADS)

Hashi, Kenjiro; Shimizu, Tadashi; Goto, Atsushi; Kiyoshi, Tsukasa; Matsumoto, Shinji; Wada, Hitoshi; Fujito, Teruaki; Hasegawa, Ken-ichi; Yoshikawa, Masatoshi; Miki, Takashi; Ito, Satoshi; Hamada, Mamoru; Hayashi, Seiji

2002-06-01

We have developed a 920-MHz NMR system and performed the proton NMR measurement of H 2O and ethylbenzene using the superconducting magnet operating at 21.6 T (920 MHz for proton), which is the highest field produced by a superconducting NMR magnet in the persistent mode. From the NMR measurements, it is verified that both homogeneity and stability of the magnet have a specification sufficient for a high resolution NMR.

Periodic harvesting of embryonic stem cells from a hollow-fiber membrane based four-compartment bioreactor.

PubMed

Knöspel, Fanny; Freyer, Nora; Stecklum, Maria; Gerlach, Jörg C; Zeilinger, Katrin

2016-01-01

Different types of stem cells have been investigated for applications in drug screening and toxicity testing. In order to provide sufficient numbers of cells for such in vitro applications a scale-up of stem cell culture is necessary. Bioreactors for dynamic three-dimensional (3D) culture of growing cells offer the option for culturing large amounts of stem cells at high densities in a closed system. We describe a method for periodic harvesting of pluripotent stem cells (PSC) during expansion in a perfused 3D hollow-fiber membrane bioreactor, using mouse embryonic stem cells (mESC) as a model cell line. A number of 100 × 10(6) mESC were seeded in bioreactors in the presence of mouse embryonic fibroblasts (MEF) as feeder cells. Over a cultivation interval of nine days cells were harvested by trypsin perfusion and mechanical agitation every second to third culture day. A mean of 380 × 10(6) mESC could be removed with every harvest. Subsequent to harvesting, cells continued growing in the bioreactor, as determined by increasing glucose consumption and lactate production. Immunocytochemical staining and mRNA expression analysis of markers for pluripotency and the three germ layers showed a similar expression of most markers in the harvested cells and in mESC control cultures. In conclusion, successful expansion and harvesting of viable mESC from bioreactor cultures with preservation of sterility was shown. The present study is the first one showing the feasibility of periodic harvesting of adherent cells from a continuously perfused four-compartment bioreactor including further cultivation of remaining cells. © 2015 American Institute of Chemical Engineers.

Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes

PubMed Central

Watson, Dionysios C.; Yung, Bryant C.; Bergamaschi, Cristina; Chowdhury, Bhabadeb; Bear, Jenifer; Stellas, Dimitris; Morales-Kastresana, Aizea; Jones, Jennifer C.; Felber, Barbara K.; Chen, Xiaoyuan; Pavlakis, George N.

2018-01-01

ABSTRACT The development of extracellular vesicles (EV) for therapeutic applications is contingent upon the establishment of reproducible, scalable, and high-throughput methods for the production and purification of clinical grade EV. Methods including ultracentrifugation (U/C), ultrafiltration, immunoprecipitation, and size-exclusion chromatography (SEC) have been employed to isolate EV, each facing limitations such as efficiency, particle purity, lengthy processing time, and/or sample volume. We developed a cGMP-compatible method for the scalable production, concentration, and isolation of EV through a strategy involving bioreactor culture, tangential flow filtration (TFF), and preparative SEC. We applied this purification method for the isolation of engineered EV carrying multiple complexes of a novel human immunostimulatory cytokine-fusion protein, heterodimeric IL-15 (hetIL-15)/lactadherin. HEK293 cells stably expressing the fusion cytokine were cultured in a hollow-fibre bioreactor. Conditioned medium was collected and EV were isolated comparing three procedures: U/C, SEC, or TFF + SEC. SEC demonstrated comparable particle recovery, size distribution, and hetIL-15 density as U/C purification. Relative to U/C, SEC preparations achieved a 100-fold reduction in ferritin concentration, a major protein-complex contaminant. Comparative proteomics suggested that SEC additionally decreased the abundance of cytoplasmic proteins not associated with EV. Combination of TFF and SEC allowed for bulk processing of large starting volumes, and resulted in bioactive EV, without significant loss in particle yield or changes in size, morphology, and hetIL-15/lactadherin density. Taken together, the combination of bioreactor culture with TFF + SEC comprises a scalable, efficient method for the production of highly purified, bioactive EV carrying hetIL-15/lactadherin, which may be useful in targeted cancer immunotherapy approaches. PMID:29535850

Quantitative Validation of the Presto Blue Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System.

PubMed

Sonnaert, Maarten; Papantoniou, Ioannis; Luyten, Frank P; Schrooten, Jan Ir

2015-06-01

As the fields of tissue engineering and regenerative medicine mature toward clinical applications, the need for online monitoring both for quantitative and qualitative use becomes essential. Resazurin-based metabolic assays are frequently applied for determining cytotoxicity and have shown great potential for monitoring 3D bioreactor-facilitated cell culture. However, no quantitative correlation between the metabolic conversion rate of resazurin and cell number has been defined yet. In this work, we determined conversion rates of Presto Blue, a resazurin-based metabolic assay, for human periosteal cells during 2D and 3D static and 3D perfusion cultures. Our results showed that for the evaluated culture systems there is a quantitative correlation between the Presto Blue conversion rate and the cell number during the expansion phase with no influence of the perfusion-related parameters, that is, flow rate and shear stress. The correlation between the cell number and Presto Blue conversion subsequently enabled the definition of operating windows for optimal signal readouts. In conclusion, our data showed that the conversion of the resazurin-based Presto Blue metabolic assay can be used as a quantitative readout for online monitoring of cell proliferation in a 3D perfusion bioreactor system, although a system-specific validation is required.

Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

PubMed Central

Sonnaert, Maarten; Papantoniou, Ioannis; Luyten, Frank P.

2015-01-01

As the fields of tissue engineering and regenerative medicine mature toward clinical applications, the need for online monitoring both for quantitative and qualitative use becomes essential. Resazurin-based metabolic assays are frequently applied for determining cytotoxicity and have shown great potential for monitoring 3D bioreactor-facilitated cell culture. However, no quantitative correlation between the metabolic conversion rate of resazurin and cell number has been defined yet. In this work, we determined conversion rates of Presto Blue™, a resazurin-based metabolic assay, for human periosteal cells during 2D and 3D static and 3D perfusion cultures. Our results showed that for the evaluated culture systems there is a quantitative correlation between the Presto Blue conversion rate and the cell number during the expansion phase with no influence of the perfusion-related parameters, that is, flow rate and shear stress. The correlation between the cell number and Presto Blue conversion subsequently enabled the definition of operating windows for optimal signal readouts. In conclusion, our data showed that the conversion of the resazurin-based Presto Blue metabolic assay can be used as a quantitative readout for online monitoring of cell proliferation in a 3D perfusion bioreactor system, although a system-specific validation is required. PMID:25336207

PERFORMANCE OF NORTH AMERICAN BIOREACTOR LANDFILLS: I. LEACHATE HYDROLOGY AND WASTE SETTLEMENT

EPA Science Inventory

An assessment of state-of-the-practice at five full-scale North American landfills operating as bioreactors is presented in this two-paper set. This paper focuses on effectiveness of liners and leachate collection systems, leachate generation rates, leachate recirculation practi...

Live imaging flow bioreactor for the simulation of articular cartilage regeneration after treatment with bioactive hydrogel.

PubMed

Bar, Assaf; Ruvinov, Emil; Cohen, Smadar

2018-06-05

Osteochondral defects (OCDs) are conditions affecting both cartilage and the underlying bone. Since cartilage is not spontaneously regenerated, our group has recently developed a strategy of injecting bioactive alginate hydrogel into the defect for promoting endogenous regeneration of cartilage via presentation of affinity-bound transforming growth factor β1 (TGF-β1). As in vivo model systems often provide only limited insights as for the mechanism behind regeneration processes, here we describe a novel flow bioreactor for the in vitro modeling of the OCD microenvironment, designed to promote cell recruitment from the simulated bone marrow compartment into the hydrogel, under physiological flow conditions. Computational fluid dynamics modeling confirmed that the bioreactor operates in a relevant slow-flowing regime. Using a chemotaxis assay, it was shown that TGF-β1 does not affect human mesenchymal stem cell (hMSC) chemotaxis in 2D culture. Accessible through live imaging, the bioreactor enabled monitoring and discrimination between erosion rates and profiles of different alginate hydrogel compositions, using green fluorescent protein-expressing cells. Mathematical modeling of the erosion front progress kinetics predicted the erosion rate in the bioreactor up to 7 days postoperation. Using quantitative real-time polymerase chain reaction of early chondrogenic markers, the onset of chondrogenic differentiation in hMSCs was detected after 7 days in the bioreactor. In conclusion, the designed bioreactor presents multiple attributes, making it an optimal device for mechanistical studies, serving as an investigational tool for the screening of other biomaterial-based, tissue engineering strategies. © 2018 Wiley Periodicals, Inc.

A miniaturized, optically accessible bioreactor for systematic 3D tissue engineering research.

PubMed

Laganà, Matteo; Raimondi, Manuela T

2012-02-01

Perfusion bioreactors are widely used in tissue engineering and pharmaceutical research to provide reliable models of tissue growth under controlled conditions. Destructive assays are not able to follow the evolution of the growing tissue on the same construct, so it is necessary to adopt non-destructive analysis. We have developed a miniaturized, optically accessible bioreactor for interstitial perfusion of 3D cell-seeded scaffolds. The scaffold adopted was optically transparent, with highly defined architecture. Computational fluid dynamics (CFD) analysis was useful to predict the flow behavior in the bioreactor scaffold chamber (that was laminar flow, Re = 0.179, with mean velocity equal to 100 microns/s). Moreover, experimental characterization of the bioreactor performance gave that the maximum allowable pressure was 0.06 MPa and allowable flow rate up to 25 ml/min. A method, to estimate quantitatively and non destructively the cell proliferation (from 15 to 43 thousand cells) and tissue growth (from 2% to 43%) during culture time, was introduced and validated. An end point viability test was performed to check the experimental set-up overall suitability for cell culture with successful results. Morphological analysis was performed at the end time point to show the complex tridimensional pattern of the biological tissue growth. Our system, characterized by controlled conditions in a wide range of allowable flow rate and pressure, permits to systematically study the influence of several parameters on engineered tissue growth, using viable staining and a standard fluorescence microscope.

Evaluation of oxygen transfer rates in stirred-tank bioreactors for clinical manufacturing.

PubMed

Bellucci, Joseph J; Hamaker, Kent H

2011-01-01

Several methods are available for determining the volumetric oxygen transfer coefficient in bioreactors, though their application in industrial bioprocess has been limited. To be practically useful, mass transfer measurements made in nonfermenting systems must be consistent with observed microbial respiration rates. This report details a procedure for quantifying the relationship between agitation frequency and oxygen transfer rate that was applied in stirred-tank bioreactors used for clinical biologics manufacturing. The intrinsic delay in dissolved oxygen (DO) measurement was evaluated by shifting the bioreactor pressure and fitting a first-order mathematical model to the DO response. The dynamic method was coupled with the DO lag results to determine the oxygen transfer rate in Water for Injection (WFI) and a complete culture medium. A range of agitation frequencies was investigated at a fixed air sparge flow rate, replicating operating conditions used in Pichia pastoris fermentation. Oxygen transfer rates determined by this method were in excellent agreement with off-gas calculations from cultivation of the organism (P = 0.1). Fermentation of Escherichia coli at different operating parameters also produced respiration rates that agreed with the corresponding dynamic method results in WFI (P = 0.02). The consistency of the dynamic method results with the off-gas data suggests that compensation for the delay in DO measurement can be combined with dynamic gassing to provide a practical, viable model of bioreactor oxygen transfer under conditions of microbial fermentation. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Micro-Macro Compatibility: When Does a Complex Systems Approach Strongly Benefit Science Learning?

ERIC Educational Resources Information Center

Samon, Sigal; Levy, Sharona T.

2017-01-01

The study explores how a complexity approach empowers science learning. A complexity approach represents systems as many interacting entities. The construct of micro-macro compatibility is introduced, the degree of similarity between behaviors at the micro- and macro-levels of the system. Seventh-grade students' learning about gases was studied…

Cell Cycle Progression of Human Cells Cultured in Rotating Bioreactor

NASA Technical Reports Server (NTRS)

Parks, Kelsey

2009-01-01

Space flight has been shown to alter the astronauts immune systems. Because immune performance is complex and reflects the influence of multiple organ systems within the host, scientists sought to understand the potential impact of microgravity alone on the cellular mechanisms critical to immunity. Lymphocytes and their differentiated immature form, lymphoblasts, play an important and integral role in the body's defense system. T cells, one of the three major types of lymphocytes, play a central role in cell-mediated immunity. They can be distinguished from other lymphocyte types, such as B cells and natural killer cells by the presence of a special receptor on their cell surface called T cell receptors. Reported studies have shown that spaceflight can affect the expression of cell surface markers. Cell surface markers play an important role in the ability of cells to interact and to pass signals between different cells of the same phenotype and cells of different phenotypes. Recent evidence suggests that cell-cycle regulators are essential for T-cell function. To trigger an effective immune response, lymphocytes must proliferate. The objective of this project is to investigate the changes in growth of human cells cultured in rotating bioreactors and to measure the growth rate and the cell cycle distribution for different human cell types. Human lymphocytes and lymphoblasts will be cultured in a bioreactor to simulate aspects of microgravity. The bioreactor is a cylindrical culture vessel that incorporates the aspects of clinostatic rotation of a solid fluid body around a horizontal axis at a constant speed, and compensates gravity by rotation and places cells within the fluid body into a sustained free-fall. Cell cycle progression and cell proliferation of the lymphocytes will be measured for a number of days. In addition, RNA from the cells will be isolated for expression of genes related in cell cycle regulations.

Bioreactor-based bone tissue engineering: The influence of dynamic flow on osteoblast phenotypic expression and matrix mineralization

PubMed Central

Yu, Xiaojun; Botchwey, Edward A.; Levine, Elliot M.; Pollack, Solomon R.; Laurencin, Cato T.

2004-01-01

An important issue in tissue engineering concerns the possibility of limited tissue ingrowth in tissue-engineered constructs because of insufficient nutrient transport. We report a dynamic flow culture system using high-aspect-ratio vessel rotating bioreactors and 3D scaffolds for culturing rat calvarial osteoblast cells. 3D scaffolds were designed by mixing lighter-than-water (density, <1g/ml) and heavier-than-water (density, >1g/ml) microspheres of 85:15 poly(lactide-co-glycolide). We quantified the rate of 3D flow through the scaffolds by using a particle-tracking system, and the results suggest that motion trajectories and, therefore, the flow velocity around and through scaffolds in rotating bioreactors can be manipulated by varying the ratio of heavier-than-water to lighter-than-water microspheres. When rat primary calvarial cells were cultured on the scaffolds in bioreactors for 7 days, the 3D dynamic flow environment affected bone cell distribution and enhanced cell phenotypic expression and mineralized matrix synthesis within tissue-engineered constructs compared with static conditions. These studies provide a foundation for exploring the effects of dynamic flow on osteoblast function and provide important insight into the design and optimization of 3D scaffolds suitable in bioreactors for in vitro tissue engineering of bone. PMID:15277663

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.

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...

Development of a Bioreactor to Culture Tissue Engineered Ureters Based on the Application of Tubular OPTIMAIX 3D Scaffolds.

PubMed

Seifarth, Volker; Gossmann, Matthias; Janke, Heinz Peter; Grosse, Joachim O; Becker, Christoph; Heschel, Ingo; Artmann, Gerhard M; Temiz Artmann, Aysegül

2015-01-01

Regenerative medicine, tissue engineering and biomedical research give hope to many patients who need bio-implants. Tissue engineering applications have already been developed based on bioreactors. Physiological ureter implants, however, do not still function sufficiently, as they represent tubular hollow structures with very specific cellular structures and alignments consisting of several cell types. The aim of this study was to a develop a new bioreactor system based on seamless, collagenous, tubular OPTIMAIX 3D prototype sponge as scaffold material for ex-vivo culturing of a tissue engineered ureter replacement for future urological applications. Particular emphasis was given to a great extent to mimic the physiological environment similar to the in vivo situation of a ureter. NIH-3T3 fibroblasts, C2C12, Urotsa and primary genitourinary tract cells were applied as co-cultures on the scaffold and the penetration of cells into the collagenous material was followed. By the end of this study, the bioreactor was functioning, physiological parameter as temperature and pH and the newly developed BIOREACTOR system is applicable to tubular scaffold materials with different lengths and diameters. The automatized incubation system worked reliably. The tubular OPTIMAIX 3D sponge was a suitable scaffold material for tissue engineering purposes and co-cultivation procedures. © 2015 S. Karger AG, Basel.

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.

Characteristics, Process Parameters, and Inner Components of Anaerobic Bioreactors

PubMed Central

Abdelgadir, Awad; Chen, Xiaoguang; Liu, Jianshe; Xie, Xuehui; Zhang, Jian; Zhang, Kai; Wang, Heng; Liu, Na

2014-01-01

The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive. PMID:24672798

Characteristics, process parameters, and inner components of anaerobic bioreactors.

PubMed

Abdelgadir, Awad; Chen, Xiaoguang; Liu, Jianshe; Xie, Xuehui; Zhang, Jian; Zhang, Kai; Wang, Heng; Liu, Na

2014-01-01

The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive.

speaq 2.0: A complete workflow for high-throughput 1D NMR spectra processing and quantification.

PubMed

Beirnaert, Charlie; Meysman, Pieter; Vu, Trung Nghia; Hermans, Nina; Apers, Sandra; Pieters, Luc; Covaci, Adrian; Laukens, Kris

2018-03-01

Nuclear Magnetic Resonance (NMR) spectroscopy is, together with liquid chromatography-mass spectrometry (LC-MS), the most established platform to perform metabolomics. In contrast to LC-MS however, NMR data is predominantly being processed with commercial software. Meanwhile its data processing remains tedious and dependent on user interventions. As a follow-up to speaq, a previously released workflow for NMR spectral alignment and quantitation, we present speaq 2.0. This completely revised framework to automatically analyze 1D NMR spectra uses wavelets to efficiently summarize the raw spectra with minimal information loss or user interaction. The tool offers a fast and easy workflow that starts with the common approach of peak-picking, followed by grouping, thus avoiding the binning step. This yields a matrix consisting of features, samples and peak values that can be conveniently processed either by using included multivariate statistical functions or by using many other recently developed methods for NMR data analysis. speaq 2.0 facilitates robust and high-throughput metabolomics based on 1D NMR but is also compatible with other NMR frameworks or complementary LC-MS workflows. The methods are benchmarked using a simulated dataset and two publicly available datasets. speaq 2.0 is distributed through the existing speaq R package to provide a complete solution for NMR data processing. The package and the code for the presented case studies are freely available on CRAN (https://cran.r-project.org/package=speaq) and GitHub (https://github.com/beirnaert/speaq).

speaq 2.0: A complete workflow for high-throughput 1D NMR spectra processing and quantification

PubMed Central

Pieters, Luc; Covaci, Adrian

2018-01-01

Nuclear Magnetic Resonance (NMR) spectroscopy is, together with liquid chromatography-mass spectrometry (LC-MS), the most established platform to perform metabolomics. In contrast to LC-MS however, NMR data is predominantly being processed with commercial software. Meanwhile its data processing remains tedious and dependent on user interventions. As a follow-up to speaq, a previously released workflow for NMR spectral alignment and quantitation, we present speaq 2.0. This completely revised framework to automatically analyze 1D NMR spectra uses wavelets to efficiently summarize the raw spectra with minimal information loss or user interaction. The tool offers a fast and easy workflow that starts with the common approach of peak-picking, followed by grouping, thus avoiding the binning step. This yields a matrix consisting of features, samples and peak values that can be conveniently processed either by using included multivariate statistical functions or by using many other recently developed methods for NMR data analysis. speaq 2.0 facilitates robust and high-throughput metabolomics based on 1D NMR but is also compatible with other NMR frameworks or complementary LC-MS workflows. The methods are benchmarked using a simulated dataset and two publicly available datasets. speaq 2.0 is distributed through the existing speaq R package to provide a complete solution for NMR data processing. The package and the code for the presented case studies are freely available on CRAN (https://cran.r-project.org/package=speaq) and GitHub (https://github.com/beirnaert/speaq). PMID:29494588

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.