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Screening and Identification of Cryopreservative Agents for Human Cellular Biotechnology Experiments in MicrogravityDimethyl sulfoxide (DMSO) has been used as a standard cryopreservative agent for mammalian cell culture; however, prolonged exposure of thawed cells to DMSO can alter cell growth. While DMSO is easily eliminated in ground-based experiments, removal of DMSO in flight-based experiments is more difficult due to various on-orbit constraints. Failure of cryopreservation is due to a number of factors, including intracellular ice formation, solute effect, and apoptotic cell death following thawing. One objective of this study is to identify and characterize an alternative cryopreservative that could be used on the International Space Station (ISS). We systematically screened for potential permeating and non-permeating agents using a human colorectal carcinoma cell line, MIP-101. Cells were suspended in cryopreservation solution and frozen either following a two-step procedure involving initial cooling at -1 C/min overnight followed by storage in liquid nitrogen (LN2) vapor, or by freezing cells directly in the LN2 vapor phase at -10 C/min. Ability to preserve cellular function after one cycle of freeze-thawing was assessed by the recovery of viable cells in short and long-term cell culture experiments. Results showed that permeating preservatives glycerol (G) and ethylene glycol (EG) had an efficacy (80-110%) comparable to, if not better than, 7.5% DMSO; but, propylene glycol (PG) had a somewhat lesser efficacy. Among the non-permeating preservatives, trehalose, raffinose, and dextran exhibited significant protective effect (50-80%) relative to that offered by 7.5% DMSO, but at -10 C and not at -1 C/min cooling rate. Preliminary data thus suggest that a combination of permeating and non-permeating agents may have improved efficacy as a cryoprotectant and serve as an alternate to DMSO for experimentation on ISS.
Document ID
20060020174
Acquisition Source
Johnson Space Center
Document Type
Conference Paper
Authors
Love,J.
(NASA Johnson Space Center Houston, TX, United States)
Elliott, T.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Das, G. C.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Hammond, D. K.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Schwarzkopf, R. J.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Jones, L. B.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Baker, T. L.
(Wyle Life Sciences, Inc. Houston, TX, United States)
Date Acquired
August 23, 2013
Publication Date
January 1, 2006
Subject Category
Exobiology
Meeting Information
Meeting: 2004 ASGSB Meeting
Location: Brooklyn, NY,
Country: United States
Start Date: November 9, 2004
End Date: November 12, 2004
Sponsors: American Society for Gravitational and Space Biology
Funding Number(s)
CONTRACT_GRANT: NAS9-02078
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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