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The fifth day of the STS-70 Space Shuttle Discovery mission is contained on this video. The crew continues working on experiments, such as the Space Tissue Loss Analysis and the Bioreactor Development System. CNN reporter, John Holliman, interviewed the flight crew and the crew also answered questions posed by Internet users while on NASA's Shuttle Web. There are brief views of Earth's surface included.
In this post-flight overview, the flight crew of the STS-70 mission, Tom Henricks (Cmdr.), Kevin Kregel (Pilot), Major Nancy Currie (MS), Dr. Mary Ellen Weber (MS), and Dr. Don Thomas (MS), discuss their mission and accompanying experiments. Pre-flight, launch, and orbital footage is followed by the in-orbit deployment of the Tracking and Data Relay Satellite (TDRS) and a discussion of the following spaceborne experiments: a microgravity bioreactor experiment to grow 3D body-like tissue; pregnant rat muscular changes in microgravity; embryonic development in microgravity; Shuttle Amateur Radio Experiment (SAREX); terrain surface imagery using the HERCULES camera; and a range of other physiological tests, including an eye and vision test. Views of Earth include: tropical storm Chantal; the Nile River and Red Sea; lightning over Brazil. A three planet view (Earth, Mars, and Venus) was taken right before sunrise. The end footage shows shuttle pre-landing checkout, entry, and landing, along with a slide presentation of the flight.
The STS-44 mission is highlighted in this first part of a two part video set. The flight crew consisted of: Cmdr. Fred Gregory; Pilot Tom Hendricks; Payload Specialist Tom Hennen; and Mission Specialists Story Musgrave, Jim Voss, and Mario Runco. The primary space shuttle mission objective was the deployment of the Defense Support Program (DSP) satellite. Secondary payload and spaceborne experiments consisted of a microbial air sampler, the Terra Scout PADVOS system, an M88-1 camera demonstration, a lower body negative pressure test, the Visual Function Tester, and a bioreactor demonstration. A tour of the flight deck, mid-deck, bathroom, and flight compartments with explanations of the equipment found in each area was conducted, a trash compactor was demonstrated, and footage of the crew together for their Thanksgiving dinner was shown. Earth views include several oceans, cloud cover, typhoon Yuri, northeast Australia, and the Barrier Reef Islands. The actor John Patrick Stewart (Commander Pickard of the show 'Star Trek: The Next Generation') performed the wake-up call for the astronauts. This flight was shortened due to an inertial measurement unit failure on the sixth day of the mission.
The highlights of the STS-70 mission are presented in this video. The flight crew consisted of Cmdr. Tom Henricks, Pilot Kevin Kregel, Flight Engineer Nancy Currie, and Mission Specialists Dr. Don Thomas and Dr. Mary Ellen Weber. The mission's primary objective was the deployment of the 7th Tracking Data and Relay Satellite (TDRS), which will provide a communication, tracking, telemetry, data acquisition, and command services space-based network system essential to low Earth orbital spacecraft. Secondary mission objectives included activating and studying the Physiological and Anatomical Rodent Experiment/National Institutes of Health-Rodents (PARE/NIH-R), The Bioreactor Demonstration System (BDS), the Commercial Protein Crystal Growth (CPCG) studies, the Space Tissue Loss/National Institutes of Health-Cells (STL/NIH-C) experiment, the Biological Research in Canisters (BRIC) experiment, Shuttle Amateur Radio Experiment-2 (SAREX-2), the Visual Function Tester-4 (VFT-4), the Hand-Held, Earth Oriented, Real-Time, Cooperative, User-Friendly, Location-Targeting and Environmental System (HERCULES), the Microcapsules in Space-B (MIS-B) experiment, the Windows Experiment (WINDEX), the Radiation Monitoring Equipment-3 (RME-3), and the Military Applications of Ship Tracks (MAST) experiment. There was an in-orbit dedication ceremony by the spacecrew and the newly Integrated Mission Control Center to commemorate the Center's integration. The STS-70 mission was the first mission monitored by this new control center. Earth views included the Earth's atmosphere, a sunrise over the Earth's horizon, several views of various land masses, some B/W lightning shots, some cloud cover, and a tropical storm.
MIP-101 is a poorly differentiated human colon carcinoma cell line established from ascites that produces minimal amounts of carcinoembryonic antigen (CEA), a 180 kDa glycoprotein tumor marker, and nonspecific cross-reacting antigen (NCA), a related protein that has 50 and 90 kDa isoforms, in vitro in monolayer culture. MIP-101 produces CEA when implanted into the peritoneum of nude mice but not when implanted into subcutaneous tissue. We tested whether MIP-101 cells may be induced to express CEA when cultured on microcarrier beads in three-dimensional cultures, either in static cultures as non-adherent aggregates or under dynamic conditions in a NASA-designed low shear stress bioreactor. MIP- 101 cells proliferated well under all three conditions and increased CEA and NCA production 3 - 4 fold when grown in three-dimensional cultures compared to MIP-101 cells growing logarithmically in monolayers. These results suggest that three-dimensional growth in vitro simulates tumor function in vivo and that three-dimensional growth by itself may enhance production of molecules that are associated with the metastatic process.
Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and therefore provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75 % satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D flat culture and 3-D HARV culture. Plating efficiency (cells attached - cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and non-satellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability since glucose levels in media from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARVS were joined together by cells into three-dimensional aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a three dimensional level of organization that could provide a more suitable model to study postnatal muscle development.
In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by non-myocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA designed High-Aspect-Ratio-Vessel (HARV) bioreactors provide a low shear environment which allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells in cultured in HARV's adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARV's using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar, however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissue-like organizations of cardiac cells in simulated microgravity.
The STS-70 Space Shuttle Program Mission Report summarizes the Payload activities as well as the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Reusable Solid Rocket Motor (RSRM), and the Space Shuttle main engine (SSME) systems performance during the seventieth flight of the Space Shuttle Program, the forty-fifth flight since the return-to-flight, and the twenty-first flight of the Orbiter Discovery (OV-103). In addition to the Orbiter, the flight vehicle consisted of an ET that was designated ET-71; three SSMEs that were designated as serial numbers 2036, 2019, and 2017 in positions 1, 2, and 3, respectively; and two SRBs that were designated 81-073. The RSRMs, designated RSRM-44, were installed in each SRB and were designated as 36OL044A for the left SRB, and 36OL044B for the right SRB. The primary objective of this flight was to deploy the Tracking and Data Relay Satellite-G/Inertial Upper Stage (TDRS-G/IUS). The secondary objectives were to fulfill the requirements of the Physiological and Anatomical Rodent Experiment/National Institutes of Health-Rodents (PARE/NIH-R); Bioreactor Demonstration System (BDS); Commercial Protein Crystal Growth (CPCG) experiment; Space Tissue Loss/National Institutes of Health - Cells (STL/NIH-C) experiment; Biological Research in Canisters (BRIC) experiment; Shuttle Amateur Radio Experiment-2 (SAREX-2); Visual Function Tester-4 (VFT-4); Hand-Held, Earth-Oriented, Real-Time, Cooperative, User-Friendly Location-Targeting and Environmental System (HERCULES); Microencapsulation in Space-B (MIS-B) experiment; Window Experiment (WINDEX); Radiation Monitoring Equipment-3 (RME-3); and the Military Applications of Ship Tracks (MAST) payload.
A variety of technologies were analyzed for their potential to remove ethylene from the CELSS Biomass Production Chamber (BPC). During crop production (e.g., lettuce, wheat, soybean, potato) in the BPC ethylene can accumulate in the airspace and subsequently affect plant viability. The chief source of ethylene is the plants themselves which reside in plastic trays containing nutrient solution. The main sink for ethylene is chamber leakage. The removal technology can be employed when deleterious levels (e.g., 50 ppb for potato) of ethylene are exceeded in the BPC and perhaps to optimize the plant growth process once a better understanding is developed of the relationship between exogenous ethylene concentration and plant growth. The technologies examined were catalytic oxidation, molecular sieve, cryotrapping, permanganate absorption, and UV degradation. Upon analysis, permanganate was chosen as the most suitable method. Experimental data for ethylene removal by permanganate during potato production was analyzed in order to design a system for installation in the BPC air duct. In addition, an analysis of the impact on ethylene concentration in the BPC of integrating the Breadboard Scale Aerobic Bioreactor (BSAB) with the BPC was performed. The result indicates that this unit has no significant effect on the ethylene material balance as a source or sink.
Fluid dynamic experiments were flown on STS-70 as phase two of the engineering evaluation of the first bioreactor Engineering Development Unit (EDU#1). The phase one experiments were comparative cell cultures in identical units on earth and onboard STS-70. In phase two, two types of fluid dynamic experiments were performed. Qualitative comparisons of the basic flow patterns were evaluated with the use of 'dye' streaklines formed from alternate injections of either a mild acid or base solution into the external flow loop that was then perfused into the vessel. The presence of Bromothymol Blue in the fluid then caused color changes from yellow to blue or vice versa, indicating the basic fluid motions. This reversible change could be repeated as desired. In the absence of significant density differences in the fluid, the flow patterns in space should be the same as on earth. Video tape records of the flow patterns for a wide range of operating conditions were obtained. The second type of fluid dynamic experiment was the quantitative evaluation of the trajectories of solid beads of various densities and sizes. The beads were introduced into the vessel and the paths recorded on video tape, with the vessel operated at various rotation rates and flow perfusion rates. Because of space limitations, the video camera was placed as close as possible to the vessel, resulting in significant optical distortion. This report describes the analysis methods to obtain comparisons between the in-flight fluid dynamics and numerical models of the flow field. The methods include optical corrections to the video images and calculation of the bead trajectories for given operating conditions and initial bead locations.
Various aspects of spaceflight, including microgravity, cosmic radiation, and physiological stress, may perturb immune function. We sought to understand the impact of microgravity alone on the cellular mechanisms critical to immunity. We utilized clinostatic RWV bioreactors that simulate aspects of microgravity to analyze the response of human PBMC to polyclonal activation. PHA responsiveness in the RWV was almost completely diminished. IL-2 and IFN-gamma secretion was reduced whereas IL- 1 beta and IL-6 secretion was increased, suggesting that monocytes may not be as adversely affected by simulated microgravity as T cells. Activation marker expression (CD25, CD69, CD71) was significantly reduced in RWV cultures. Furthermore, addition of exogenous IL-2 to these cultures did not restore proliferation. Reduced cell-cell and cell-substratum interactions may play a role in the loss of PHA responsiveness. However, PHA activation in Teflon culture bags that limit cell-substratum interactions did not suppress PHA activation. Furthermore, increasing cell density and, therefore, cell-cell interactions in the RWV cultures did not help restore PHA activation. However, placing PBMC within small collagen beads did partially restore PHA responsiveness. Activation of both PBMC and purified T cells with PMA and ionomycin was unaffected by RWV culture, indicating that signaling mechanisms downstream of PKC activation and calcium flux are not sensitive to simulated microgravity. Furthermore, submitogenic doses of PMA alone but not ionomycin alone restored PHA responsiveness of PBMC in RWV culture. Thus, our data indicate that during polyclonal activation the signaling pathways upstream of PKC activation are sensitive to simulated microgravity.
The objective of the research was to begin evaluating the effect of zero-shear on the development of the cell wall of Saccharomyces cerevisiae employing the High Aspect Rotating-Wall Vessel (HARV) NASA bioreactor. This particular yeast has enormous potential for research as a model eukaryotic system on the International Space Station, as well as the production of food stuffs' at the future lunar colony. Because the cell wall is the barrier between the cell and the environment, its form and function as influenced by microgravity is of great importance. Morphologic studies revealed that the circularity and total area of the individual yeast cells were essentially the same in both the control and test HARV's. The growth rates were also essentially the same. In zero-shear, the yeast grew in clumps consisting of rudimentary pseudohyphae in contrast to solitary budding cells in the control. Based upon mechanical and sonic shear applied to the yeast cells, those grown in zero-shear had stronger cell walls and septa. This suggests that there are structural differences, most likely related to the chitin skeleton of the cell wall. From this research further NASA support was obtained to continue the work. Investigations will deal with gene expression and ultrastructure. These will lead to a clearer assessment of the value of S. cerevisiae eukaryotic as a model for space station research.
Attempts to simulate normal tissue micro-environments in vitro have been thwarted by the complexity and plasticity of the extracellular matrix, which is important in regulating cytoskeletal and nuclear matrix proteins. Gravity is one of the problems, tending to separate components that should be kept together. For space shuttle experiments, NASA engineers devised a double-walled rotating bioreactor, which is proving to be a useful tissue culture device on earth as well as in space.
On this sixth day of the STS-85 mission, the flight crew, Cmdr. Curtis L. Brown, Jr., Pilot Kent V. Rominger, Payload Cmdr. N. Jan Davis (Ph.D.), Mission Specialists Robert L. Curbeam, Jr. and Stephen K. Robinson (Ph.D.), and Payload Specialist Bjarni V. Tryggvason today continue their work with the Bioreactor Demonstration System designed to perform cell biology experiments under controlled conditions. Tryggvason, today continues his work with the Microgravity Vibration Isolation Mount which uses magnets to levitate a platform and protect sensitive microgravity processing experiments from vibrations.
On this fifth day of the STS-85 mission, the flight crew, Cmdr. Curtis L. Brown, Jr., Pilot Kent V. Rominger, Payload Cmdr. N. Jan Davis (Ph.D.), Mission Specialists Robert L. Curbeam, Jr. and Stephen K. Robinson (Ph.D.), and Payload Specialist Bjarni V. Tryggvason once again test the small robotic arm serving as a prototype for one that will fly as part of the Japanese Experiment Module on the International Space Station. Simulated orbital replacement unit detachment and reattachment will be the focus. Bob Curbeam discusses the progress of the flight with a television station in St Louis, before continuing his work with the Bioreactor Demonstration System designed to perform cell biology experiments under controlled conditions. Immediately after Curbeam's interview, Canadian Payload Specialist Bjarni Tryggvason is set to talk to elementary and high school students at a summer camp in Saskatchewan, Canada.
On this eighth day of the STS-85 mission, the flight crew, Cmdr. Curtis L. Brown, Jr., Pilot Kent V. Rominger, Payload Cmdr. N. Jan Davis (Ph.D.), Mission Specialists Robert L. Curbeam, Jr. and Stephen K. Robinson (Ph.D.), and Payload Specialist Bjarni V. Tryggvason entered the final portion of its flight. The new Mir 24 crew of Commander Anatoly Solovyev and Flight Engineer Pavel Vinogradov, who arrived on the station the same day Discovery was launched, bid farewell to Mir 23 Commander Vasily Tsibliev and Flight Engineer Alexander Lazutkin who are returning home after 185 days in space. The Soyuz vehicle carrying the Mir 23 crew home undocked from the station. Robinson again used the Southwest Ultraviolet Imaging System (SWUIS), a 7-inch imaging telescope that is pointed out of the orbiter's middeck hatch window, to observe the Hale-Bopp comet. Curbeam continued his work with the Bioreactor Demonstration System designed to perform cell biology experiments under controlled conditions. Tryggvason spent part of his time troubleshooting a computer hard drive system that supports the Microgravity Vibration Isolation Mount experiment.
On this ninth day of the STS-85 mission, the flight crew, Cmdr. Curtis L. Brown, Jr., Pilot Kent V. Rominger, Payload Cmdr. N. Jan Davis (Ph.D.), Mission Specialists Robert L. Curbeam, Jr. and Stephen K. Robinson (Ph.D.), and Payload Specialist Bjarni V. Tryggvason watch over the Manipulator Flight Demonstration (MFD) experiment while Japanese investigators again maneuvere the Small Fine Arm remotely from a control room near Mission Control. It is the final planned work with the arm during this mission. While MFD operations are ongoing, Robinson again uses the Southwest Ultraviolet Imaging System's ultraviolet imaging telescope to observe Comet Hale-Bopp and Curbeam continue his work with the Bioreactor Demonstration System designed to perform cell biology experiments under controlled conditions. Tryggvason spends his day supporting data gathering with the Microgravity Vibration Isolation Mount experiment. Before the crew's workday began, they discussed the mission's progress with reporters in the U.S. and Canada as part of the traditional crew news conference. Questions ranged from life in space for the first time space travelers to providing a report card on the more than 24 experiments being conducted throughout the mission.
The macromolecules comprising the cytoskeleton and extracellular matrix of cells may be sensitive to gravitation. Since early development of organs depends on dynamic interactions across cell surfaces, altered gravity may disturb development. We investigated this possibility for heart development. Previous studies showed that the extracellular matrix glycoprotein fibronectin (Fn) is necessary for normal heart development. We cultured precardiac tissue explants in a high aspect ratio bioreactor vessel (HARV) to simulate microgravity. We observed tissue morphology, contraction, and Fn distribution by immunolocalization in HARV rotated and control (lxg) explants, cultured 18 hr. We also measured Fn amount by immunoassay. Explants in HARV were rotated at 6 rpm to achieve continuous freefall. Thirty-five of 37 control, but only 1 of 37 matched rotated explants exhibited contractions. Tissue architecture was identical. Immunolocalization of Fn showed remarkable differences which may be related to the development of contractions. The Fn staining in the HARV explants was less intense in all areas. Areas of linear staining along epithelia were present but shorter, and there was less intercellular staining in both mesenchymal tissue and myocardium. Initial immunoassay results of 5 matched pairs of explants showed a 22% reduction in total tissue Fn in the HARV rotated samples. Our results indicate that altered gravity in the HARV reduced the amount and distribution of Fn, as assessed by two independent criteria. This was correlated with a reduction in the development of contractile activity.