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Cellular Biotechnology Operations Support Systems: Production of Recombinant Human Erythropoietin by Mammalian Cells (CBOSS-02-Erythropoietin)
06.27.08

Overview | Description | Applications | Operations | Results | Publications | Images

Experiment/Payload Overview

Brief Summary

Cells grown in microgravity grow and replicate into complex structures, unlike cells grown on Earth. To better understand the mechanisms that cause the differentiation of cells in microgravity, seven cell lines of common human illnesses were grown on ISS. The cells were returned to Earth and were used in the studies of several human diseases. This study is important for understanding the mechanisms needed to fight immune dysfunction caused by microgravity.

Principal Investigator

  • Arthur J. Sytkowski, M.D., Ph.D., Beth Israel Deaconess Medical Center, Boston, MA

    • Co-Investigator(s)/Collaborator(s)

    No Information Available

    Payload Developer

    Wyle Laboratories, Incoporated, Houston, TX

    Sponsoring Agency

    National Aeronautics and Space Administration (NASA)

    Expeditions Assigned

    |4|

    Previous ISS Missions

    A previous version of the BSTC flew on STS-86/Mir and STS-90. BTR flew on STS-89 and STS-91. Cell-growth experiments have flown on the Shuttle since STS-70.

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    Experiment/Payload Description

    Research Summary

    • The CBOSS-02-Erythropoietin experiment is one of seven separate experiments in the CBOSS suite of experiments. The purpose of CBOSS-02-Erythropoietin is to support biotechnological research aboard ISS by providing a stable environment for growing cells.


    • This self-contained apparatus is designed to allow multiple experiments studying various types of cells to operate simultaneously. It is a multi-component cell incubator intended to grow three-dimensional clusters of cells in microgravity.


    • CBOSS-02-Erythropoietin will examine how microgravity affects the hematopoietic system (system of organs involved in the production of blood) in microgravity.

    Description

    The purpose of the Cellular Biotechnology Operations Support System (CBOSS) study was to support biotechnological research on board ISS by providing a stable environment in which to grow cells. The system was a multi-component cell incubator intended to grow three-dimensional clusters of cells in microgravity. A self-contained apparatus, CBOSS was designed to allow multiple experiments to be performed, thereby enabling scientists to study various types of cells operating simultaneously.

    In the human body, cells normally grow within a scaffolding of protein and carbohydrate fibers that creates a three dimensional structure. But outside the body, cells tend to grow in flat sheets and are incapable of duplicating the structure they normally hold, which can make them behave differently in the laboratory than they would in the body. Past research has shown that cells grown in a microgravity environment arrange themselves into three-dimensional shapes that more closely duplicate how they would behave in the body. Cell culture in microgravity thus becomes a tool for studying cells in a state that is closer to that which occurs normally in the body.

    Reduced immune system function and anemia related to decreased red blood cell production are two problems that face astronauts after extended durations in space. This experiment was designed to study cells in space to gain insight into the way microgravity affects blood cell formation. EMS-3 cells are Rauscher murine erythroleukemia cells, derived originally from mice infected with the virus that causes erythroleukemia. The EMS-3 cell line serves as an important model system for studying the cellular and molecular aspects of erythropoiesis (red blood cell formation), including the mechanism of action of erythropoietin in vitro under controlled conditions. EMS-3 cells were selected for culture in CBOSS to advance our knowledge of effects of microgravity on the hematopoietic system and to suggest possible in-flight countermeasures and treatments for ground-based disease states.

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    Applications

    Space Applications

    CBOSS-02-Erythropoietin will provide insight on how the human EMS-3 behave in a microgravity environment. This may lead to countermeasures for microgravity induced anemia during long duration space missions.

    Earth Applications

    In the human body cells normally grow within a scaffolding of protein and carbohydrate fibers that help create a three dimensional structure. This is how organs maintain their shape. Studying cells on Earth is difficult because outside the body cells tend to grow in flat sheets and are not capable of duplicating the structure they normally hold, which often makes them behave differently in the lab than they would in the body. Past research, however, has shown that cells grown in microgravity arrange themselves into three-dimensional shapes, more closely duplicating how they behave in the body. CBOSS, then, becomes very useful as a tool for studying cells in a state as close to that which occurs normally in the body.

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    Operations

    Operational Requirements

    BSTC and GSM were housed side by side in lockers 1 and 5, respectively, of EXPRESS Rack 4. At the end of Increment 3, BTR was transferred to EXPRESS Rack 4, as well. GSM does not use power or gas supplied by the EXPRESS Rack, but does interface with the Station computer via the EXPRESS Rack's Ethernet connection. BCSS-1 and -4, housed directly below BSTC and GSM in lockers 2 and 6, do not require EXPRESS Rack support. BTR is located in EXPRESS Rack 1, locker 6. It operates on 160 watts of continuous power and Ethernet connection provided by the rack.

    Operational Protocols

    After the CBOSS hardware was installed on ISS, the crew activated the experiments and monitored the status of the experiments and hardware. Crew members used a syringe to inject cells into fresh tissue culture modules (TCMs), by using the injection ports. They also added fresh media to growing cell lines and fixatives at the end of the experiment to halt growth. When the cultures were fixed, the TCMs were transferred to the BTR for storage. The crew performed periodic preventive maintenance on the CBOSS components. Video and data was downloaded to the CBOSS flight control team at the Johnson Space Center's Telescience Center. The CBOSS-02-Erythropoietin experiment's TCM was returned to the principle investigator for in-depth analysis.

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    Results/More Information

    The CBOSS hardware supported six cell culture investigations with different detailed scientific objectives. There were problems in the growth and preservation of all of the cell lines grown on Expeditions 3 and 4. The PC12 and erythroleukemia cells did not survive well in long term culture, so no scientific results are expected from these experiments. It was found that there was more bubble formation than expected that may lead to cell death at the air-liquid interface. Although not well documented in this experiment, it was noted that poor mixing of cells/tissues and medium occurred in the other CBOSS payloads as well. Both the poor mixing and greater than expected bubble formation were important lessons learned that led to the addition of the CBOSS-Fluid Dynamics Investigation (CBOSS-FDI) to study mixing and bubble formation in microgravity on later Expeditions.

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    Related Web Sites
  • NASA Fact Sheet

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    Publications

    Results Publications

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      Related Publications
      • Feldman L, Sytkowski AJ. Pleiotrophic actions of erythropoietin. Environmental Health and Preventive Medicine. ;7(6):239-245. 2003
      • Sytkowski AJ, Davis KL. Erythroid cell growth and differentiation in vitro in the simulated microgravity environment of the NASA rotating wall vessel bioreactor. In Vitro Cellular and Developmental Biology, Animal. ;37(2):79-83. 2001
      • Jessup JM, Pellis NR. NASA biotechnology: cell science in microgravity. In Vitro Cellular and Developmental Biology, Animal. ;37(2):2. 2001

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      Images

      imageStained microscopy image of erythroleukemia cells. Courtesy image from Marshall Space Flight Center.
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      imageNASA Image: ISS004E5223 - Image of a Quad Tissue Culture Module Assembly (QTCMA) 8 on ISS Expedition 4 after activation of the cells. A syringe was used to inject cells into the pink nutrient growth media. The BSTC can hold eight of these QTCMAs, which will be used to grow human cells on ISS. When the samples completed their growth cycle, the crew transfered the QTCMAs from the BSTC to the BTR, where they were stored until they are examined at a ground-based laboratory.
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      Information Provided and Updated by the ISS Program Scientist's Office
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