Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well‐controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European ‘Biorack’ provide generic experiment facilities including an incubator, on‐board 1‐g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the ‘Biorack’ facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK‐1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatent in‐flight), injection port, and supernatent collection chamber. The second unit (GCAK‐2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatent, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T‐cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground‐ based investigations simulating the conditions expected in the flight experiment. Several parameters including cell concentration, time between cell loading and activation, and storage temperature on cell survival were examined to characterise cell response and optimise the experiments to be flown aboard the Space Shuttle. Results indicate that the objectives of the experiments could be met with delays up to 5 days between cell loading into the hardware and initial in flight experiment activation, without the need for medium exchange. Experiment hardware of this kind, which is adaptable to a wide range of cell types and can be easily interfaced to different spacecraft facilities, offers the possibility for a wide range of experimenters successfully and easily to utilise future flight opportunities. J. Cell. Biochem. 70:252–267, 1998. © 1998 Wiley‐Liss, Inc.

[1]  D. Kulesh,et al.  Space shuttle flight (STS‐45) of L8 myoblast cells results in the isolation of a nonfusing cell line variant , 1994, Journal of cellular biochemistry.

[2]  Y. Nefedov,et al.  Space flight effects on Paramecium tetraurelia flown aboard Salyut 6 in the Cytos I and Cytos M experiments. , 1981, Advances in space research : the official journal of the Committee on Space Research.

[3]  A. Cogoli,et al.  Activation signals of T lymphocytes in microgravity. , 1996, Journal of biotechnology.

[4]  Augusto Cogoli,et al.  Gravitational and space biology at the cellular level , 1996 .

[5]  J. den Hertog,et al.  Nuclear responses to protein kinase C signal transduction are sensitive to gravity changes. , 1991, Experimental cell research.

[6]  A Cogoli,et al.  Cell sensitivity to gravity. , 1984, Science.

[7]  R. Stanko,et al.  Pyruvate reduces anoxic injury and free radical formation in perfused rat hepatocytes. , 1996, The American journal of physiology.

[8]  B. Spooner,et al.  Gravity in mammalian organ development: differentiation of cultured lung and pancreas rudiments during spaceflight. , 1994, The Journal of experimental zoology.

[9]  R. Ian Freshney,et al.  Culture of Animal Cells , 1983 .

[10]  Marian L. Lewis,et al.  Spaceflight alters microtubules and increases apoptosis in human lymphocytes (Jurkat) , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[11]  Augusto Cogoli,et al.  Mitogenic signal transduction in Tlymphocytes in microgravity , 1993, Journal of leukocyte biology.

[12]  S. Chapes,et al.  Abrogation of TNF-mediated cytotoxicity by space flight involves protein kinase C. , 1994, Experimental cell research.

[13]  J P Hatton,et al.  The distribution of protein kinase C in human leukocytes is altered in microgravity , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  B. Spooner,et al.  Pre-metatarsal skeletal development in tissue culture at unit- and microgravity. , 1994, The Journal of experimental zoology.

[15]  M L Lewis,et al.  Effects of microgravity on osteoblast growth activation. , 1996, Experimental cell research.

[16]  A Cogoli,et al.  Gravity effects on single cells: techniques, findings, and theory. , 1991, Advances in space biology and medicine.

[17]  K P Lee,et al.  Effect of spaceflight on human stem cell hematopoiesis: suppression of erythropoiesis and myelopoiesis , 1996, Journal of leukocyte biology.

[18]  S. Manié,et al.  Inhibition of phorbol ester-induced cell activation in microgravity. , 1991, Experimental cell research.