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At least 127 records · Page 7

Continuous-Flow Gas-Phase Bioreactors

Continuous-flow gas-phase bioreactors proposed for biochemical, food-processing, and related industries. Reactor contains one or more selected enzymes dehydrated or otherwise immobilized on solid carrier. Selected reactant gases fed into reactor, wherein chemical reactions catalyzed by enzyme(s) yield product biochemicals. Concept based on discovery that enzymes not necessarily placed in traditional aqueous environments to function as biocatalysts.

Wise, Donald L.↗

Development of a Space Bioreactor using Microtechnology

A miniature bio-reactor for the cultivation of cells aboard Spacelab is presented. Yeast cells are grown in a 3 milliliter reactor chamber. A supply of fresh nutrient medium is provided by a piezo-electric silicon micro-pump. In the reactor, pH, temperature, and redox potential are monitored and the pH is regulated at a constant value. The complete instrument is fitted in a standard experiment container of 63 x 63 x 85 mm. The bioreactor was used on the IML-2 mission in July 1994 and is being refurbished for a reflight in the spring of 1996.

Arquint, Philippe↗

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

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

Sastry, Jagannadha K.↗

Use of NASA Bioreactor in Engineering Tissue for Bone Repair

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

Duke, Pauline↗

Evaluation of the Hydrodynamic Focusing Bioreactor (HDFB) and the Centrifugal Absorption Cartridge System (CACS) Performance Under Micro G

The Hydrodynamic Focusing Bioreactor (HDFB) technology is designed to provide a flow field with nearly uniform shear force throughout the vessel, which can provide the desired low shear force spatial environment to suspend three-dimensional cell aggregates while providing optimum mass transfer. The reactor vessel consists of a dome-shaped cell culture vessel, a viscous spinner, an access port, and a rotating base. The domed vessel face has a radius of R(o). and rotates at 0mega(o) rpm, while the internal viscous spinner has a radius of R(i) and rotates at 0mega(i) rpm. The culture vessel is completely filled with cell culture medium into which three-dimensional cellular structures are introduced. The HDFB domed vessel and spinner were driven by two independent step motors,

Gonda, Steve↗

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

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

Lawless, DeSales↗

Human Bone-Forming Chondrocytes Cultured in the Hydrodynamic Focusing Bioreactor Retain Matrix Proteins: Similarities to Spaceflight Results

Fracture healing, crucial to a successful Mars mission, involves formation of a cartilaginous fracture callus which differentiates, mineralizes, ossifies and remodels via the endochondral process. Studies of spaceflown and tailsuspended rats found that, without loading, fracture callus formation and cartilage differentiation within the callus were minimal. We found delayed differentiation of chondrocytes within the rat growth plate on Cosmos 1887, 2044, and Spacelab 3. In the current study, differentiation of human bone-forming chondrocytes cultured in the hydrodynamic focusing bioreactor (HFB) was assessed. Human costochondral chondrocytes in suspension were aggregated overnight, then cultured in the HFB for 25 days. Collagen Type II, aggrecan and unsulfated chondroitin were found extracellularly and chondroitin sulfates 4 and 6 within the cell. Lack of secretion was also found in pancreatic cells of spaceflown rats, and in our SL3 studies. The HFB can be used to study cartilage differentiation in simulated microgravity.

Duke, P. J.↗

Osteocytes Mechanosensing in NASA Rotating Wall Bioreactor

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

Spatz, Jordan↗

Regulating Glucose and pH, and Monitoring Oxygen in a Bioreactor

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.

Anderson, Melody M.↗

Oxygen Sensors Monitor Bioreactors and Ensure Health and Safety

In order to cultivate healthy bacteria in bioreactors, Kennedy Space Center awarded SBIR funding to Needham Heights, Massachusetts-based Polestar Technologies Inc. to develop sensors that could monitor oxygen levels. The result is a sensor now widely used by pharmaceutical companies and medical research universities. Other sensors have also been developed, and in 2013 alone the company increased its workforce by 50 percent.

Source record↗

Fiber Treatment Effects on Bioreactor Bulk Fluid Trends

In order to facilitate the exploration of worlds beyond the borders of our planet, it is necessary to maintain sustainable levels of clean water. The remediation of water via Membrane Aerated Bioreactors (MABRs) is one such method, and the focus of this study. MARRs rely on healthy biofilms grown on hollow fiber membranes to clean non-potable water. These biofilms can take weeks to months to establish. Therefore, various fiber treatments and two inoculums were evaluated for their effect on rapid biofilm formation. Fiber treatments are as follows: sanding of the fibers with 1500 and 8000 grit sandpaper, immersion of the fibers in a 1% hydrofluoric acid solution for 12 seconds and 15 minutes, and the immersion of the fibers in a Fluoroetch® solution for 18 seconds and 5 minutes. The two inoculums utilized were sourced from healthy, established MARRs; Texas Tech University (TTU) MABR "TRL5" and Kennedy Space Center (KSC) MABR "R3". Data attained from direct bacterial cell counts of the reactor bulk fluids via fluorescent microscopy, suggests that the fluoroetching treatment combined with the TTU inoculum show the greatest biofilm creation.

Ellis, Ronald II↗

Rapid Startup and Loading of an Attached Growth, Simultaneous Nitrification/Denitrification Membrane Aerated Bioreactor

The Membrane Aerated Bioreactor (MABR) is an attached‐growth biological system for simultaneous nitrification and denitrification. This design is an innovative approach to common terrestrial wastewater treatments for nitrogen and carbon removal. Implementing a biologically‐based water treatment system for long‐duration human exploration is an attractive, low energy alternative to physiochemical processes. Two obstacles to implementing such a system are (1) the "start‐up" duration from inoculation to steady‐state operations and (2) the amount of surface area needed for the biological activity to occur. The Advanced Water Recovery Systems (AWRS) team at JSC explored these two issues through two tests; a rapid inoculation study and a wastewater loading study. Results from these tests demonstrate that the duration from inoculation to steady state can be reduced to two weeks and that the surface area to volume ratio baseline used in the Alternative Water Processor (AWP) test was higher than what was needed to remove the organic carbon and ammonium from the system.

Meyer, Caitlin↗

Bioreactor Yields Extracts for Skin Cream

Johnson Space Flight Center researchers created a unique rotating-wall bioreactor that simulates microgravity conditions, spurring innovations in drug development and medical research. Renuèll Int'l Inc., based in Aventure, Florida, licensed the technology and used it to produce a healing skin care product, RE`JUVEL. In a Food and Drug Administration test, RE`JUVEL substantially increased skin moisture and elasticity while reducing dark blotches and wrinkles.

Source record↗

Rapid Start-up and Loading of an Attached Growth, Simultaneous Nitrification/Denitrification Membrane Aerated Bioreactor

Membrane aerated bioreactors (MABR) are attached-growth biological systems used for simultaneous nitrification and denitrification to reclaim water from waste. This design is an innovative approach to common terrestrial wastewater treatments for nitrogen and carbon removal and implementing a biologically-based water treatment system for long-duration human exploration is an attractive, low energy alternative to physiochemical processes. Two obstacles to implementing such a system are (1) the "start-up" duration from inoculation to steady-state operations and (2) the amount of surface area needed for the biological activity to occur. The Advanced Water Recovery Systems (AWRS) team at JSC explored these two issues through two tests; a rapid inoculation study and a wastewater loading study. Results from these tests demonstrate that the duration from inoculation to steady state can be reduced to under two weeks, and that despite low ammonium removal rates, the MABRs are oversized.

Meyer, Caitlin E.↗

Methane Bioreactor System Offers Sustainable Plastic Alternative for Earth and Space

Mango Materials partnered with the Colorado School of Mines on NASA STTR awards to adapt a bioreactor system to convert methane into bioplastic for low-gravity environments. In space, methane from carbon dioxide conversion systems and waste treatment can be used to 3D print objects and provide polymers for construction and regolith binding. On Earth, the system can convert methane—a greenhouse gas—into environmentally conscious bioplastic. Mango Materials received $6 million through NASA CCRPP, including funds from fashion investors seeking alternatives to plastic-based textiles.

SBIR/STTR↗

A scalable and autoclavable oxygen nanosensor platform for metabolic monitoring of Saccharomyces cerevisiae in a bioreactor and other in situ systems

Polymer-encapsulated dye nanoparticle sensors are a valuable approach to achieving in situ analyte measurements with luminescence; however, typical emulsion-based nanosensors are poorly suited for large-scale biological samples due to limitations of synthesis scalability and stability. Branched polyethylenimine (PEI) is a versatile polymer scaffold ideal for constructing nanoparticles with various covalently conjugated moieties due to their high density of reactive primary amines, high water solubility, and biological stability. In this work, we used branched polyethylenimine as a scaffold-based approach for making a stable and scalable ratiometric oxygen sensor. Pt (II) tetracarboxyporphine was used as an oxygen-sensing dye and coumarin 343 as a reference dye, all covalently linked to the PEI scaffold producing a product that could withstand sterilization procedures and easily be scaled. To minimize toxicity from the PEI scaffold, we conjugated it with 2000 MW PEG. The applicability of the sensors was demonstrated in a 200 mL Saccharomyces cerevisiae yeast culture, using orthogonal luminescent and electrochemical oxygen measurements to validate sensor response and measure the metabolic activity of the yeast in our culture. Further, this approach was able to match the sensitivity of our electrochemical measurements while improving upon drawbacks of other luminescent methods of oxygen detection, demonstrating effective monitoring for at least 20 h. Our scaffold-based approach is a modular and easily translatable technology that could be useful in various biotechnological applications.

59 BASIC BIOLOGICAL SCIENCES↗

Analysis of CDC Bioreactor Internal Thermal Measurements and Sample Coupon Temperatures

The CDC Biofilm Reactor is an integral laboratory tool for the Environmental Control and Life Support Systems (ECLSS) biofilm formation and growth research program. Critical to this research is the need to adjust and maintain various surface temperatures of the coupons housed within the CDC Biofilm Reactor. The purpose of this study was to provide quantitative temperature gradient information when the CDC Biofilm Reactor was operating according to several process scenarios. Two primary process parameters were evaluated. For the first set of test parameters, the liquid level was maintained at 350 mL, with an inlet flowrate of 0.1 mL/min, 1 mL/min, 10 mL/min. The liquid was allowed to gravity drain out of the outlet spout. For the second set of test parameters, the liquid level within the reactor was maintained at 550 mL, with an inlet flow of 0.1 mL/min, 1 mL/min, 10 mL/min and draining intermittently controlled to 0.8 mL/min to maintain the 550 mL level. Due to the placement of the thermocouple in the reactor, a difference in temperature occurred between the coupon surfaces and target biofilm reactor temperature when operated according to the first set of test parameters. When the reactor was operated according to the second set of parameters, which resulted in the thermocouple being submerged, the temperature gradient was eliminated. The results demonstrated minimal temperature gradient between the top and bottom coupon surfaces for coupons placed in a single rod within the CDC Biofilm Reactor for both sets of test parameters evaluated. The collection of this information helped to explain previous ECLSS biofilm formation test runs, along with providing guidance on best operating practices for future ECLSS experiments. The placement of the thermocouple also helps to explain the challenge of achieving and maintaining bulk liquid temperatures when biofilm is grown according to the standardized test methods.

CDC Bioreactor↗