Cellular Biotechnology Operations Support Systems: PC12 Pheochromocytoma Cells - A Proven Model System for Optimizing 3-D Cell Culture Biotechnology in Space (CBOSS-01-PC12) - 07.14.16
Cells grown in microgravity grow and replicate into complex structures, unlike cells grown on Earth. To better understand the mechanisms of neural regeneration and pain suppression, a neuroendocrine cell line was grown on ISS, then returned to Earth for analysis. Science Results for Everyone
This investigation looked at neural regeneration and pain suppression using cultures of neuroendocrine (PC12) cells, which are involved in interactions of the nervous system and hormones. The cells did not survive well long-term on the International Space Station and no scientific results are expected from the experiment. However, analysis revealed poor mixing of cells and medium, and more bubble formations in the hardware than expected, which could have caused the early cell death at the air-liquid interface. Important lessons were learned from these problems, and an investigation was added to study them. Experiment Details
Peter I. Lelkes, Ph.D., Temple University, Philadelphia, PA, United States
Wyle, Integrated Science and Engineering, Houston, TX, United States
Sponsoring Space Agency
National Aeronautics and Space Administration (NASA)
Human Exploration and Operations Mission Directorate (HEOMD)
ISS Expedition Duration
August 2001 - December 2001
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.
- CBOSS-01-PC12 is one of seven in the CBOSS suite of experiments. The purpose of CBOSS 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-01-PC12 will investigate the neural regeneration and pain suppression by using neuroendocrine (PC12) cells grown onboard ISS.
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.
Neuroendocrine cells (PC12) are cells that receive electric signals from the nervous system and chemical signals secreted from glands. As they differentiate, the cells are known to produce catecholamines, which are key to normal function and pain suppression. Evidence of differentiation is seen in ground-based (simulated) microgravity rotating wall systems. In this experiment, the ability and extent of the differentiation in actual microgravity was assessed by the growth and subculture of these cells to provide a greater understanding of neural regeneration and pain suppression.
Development of techniques to reliably cultivate organisms under controlled conditions is essential to understanding the effect of microgravity and radiation on living organisms and creating environmental conditioning sources for long term space flight.
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. CBOSS-01-PC12 studies may lead to better methods of pain suppression and a greater understanding of neural regeneration.
Operational Requirements and Protocols
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.
After CBOSS was installed on ISS, the crew activated CBOSS-01-PC12 and monitored the status of the experiment and hardware. Crew members used a syringe to inject cells into fresh 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 TCM 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-01-PC12 experiment's TCM was returned to the principle investigators for in-depth analysis.
Decadal Survey Recommendations
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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. (Evans et al. 2009)^ back to top
Ground Based Results Publications
Unsworth BR, Lelkes PI. Growing tissues in microgravity. Nature Medicine. 1998; 4(8): 901-907.
Jessup JM, Pellis NR. NASA biotechnology: cell science in microgravity.. In Vitro Cellular and Developmental Biology - Animal. 2001; 37(2): 2. PMID: 11332737.
Lelkes PI, Galvan DL, Hayman GT, Goodwin TJ, Chatman DY, Cherian S, Garcia RM, Unsworth BR. Simulated microgravity conditions enhance differentiation of cultured PC12 cells towards the neuroendocrine phenotype. In Vitro Cellular and Developmental Biology - Animal. 1998; 34(4): 316-325.
NASA Fact Sheet
Stained microscopy image of neuroendocrine cells. Image courtesy of NASA, Marshall Flight Center.
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NASA Image: ISS0005309 - Image of a Quad Tissue Culture Module Assembly (QTCMA) on ISS Expedition 3 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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