Document Detail

Mechanism of cryoprotection by extracellular polymeric solutes.
MedLine Citation:
PMID:  2462928     Owner:  NLM     Status:  MEDLINE    
To elucidate the means by which polymer solutions protect cells from freezing injury, we cooled human monocytes to -80 degrees C or below in the presence of various polymers. Differential scanning calorimetric studies showed that those polymers which protect cells best have a limiting glass transition temperature (T'g) of approximately -20 degrees C; those with a T'g significantly higher or lower did not protect. Freeze-etch electron micrographs indicated that intracellular ice crystals had formed during this freezing procedure, but remained smaller than approximately 300 nm in the same proportion of cells as survived rapid thawing. We propose that cryoprotection of slowly frozen monocytes by polymers is a consequence of a T'g of -20 degrees C in the extracellular solution. In our hypothesis, the initial concentration and viscosity of protective polymer solutions reduce the extent and rate of cell water loss to extracellular ice and limit the injurious osmotic stress, which cells face during freezing at moderate rates to -20 degrees C. Below -20 degrees C, glass formation prevents further osmotic stress by isolating cells from extracellular ice crystals, virtually eliminating cell water loss at lower temperatures. On the other hand, the protective polymer solutions will allow some diffusion of water away from cells at temperatures above T'g. If conditions are correct, cells will concentrate the cytoplasm sufficiently during the initial cooling to T'g to avoid lethal intracellular freezing between T'g and the intracellular Tg, which has been depressed to low temperatures by that concentration. Thus, when polymers are used as cryoprotective agents, cell survival is contingent upon maintenance of osmotic stress within narrow limits.
T Takahashi; A Hirsh; E Erbe; R J Williams
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biophysical journal     Volume:  54     ISSN:  0006-3495     ISO Abbreviation:  Biophys. J.     Publication Date:  1988 Sep 
Date Detail:
Created Date:  1989-02-22     Completed Date:  1989-02-22     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0370626     Medline TA:  Biophys J     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  509-18     Citation Subset:  IM    
American Red Cross Holland R & D Laboratories, Rockville, Maryland.
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MeSH Terms
Blood Preservation*
Cryoprotective Agents / pharmacology*
Dextrans / pharmacology
Dimethyl Sulfoxide / pharmacology
Freeze Fracturing
Hetastarch / pharmacology
Microscopy, Electron
Monocytes / cytology,  drug effects*,  ultrastructure
Phagocytosis / drug effects
Polyethylene Glycols / pharmacology
Povidone / pharmacology
Grant Support
Reg. No./Substance:
0/Cryoprotective Agents; 0/Polyethylene Glycols; 67-68-5/Dimethyl Sulfoxide; 9003-39-8/Povidone; 9004-54-0/Dextrans; 9005-27-0/Hetastarch

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