Document Detail

Intracellular ice formation in mouse oocytes subjected to interrupted rapid cooling.
MedLine Citation:
PMID:  17686470     Owner:  NLM     Status:  MEDLINE    
The formation of ice crystals within cells (IIF) is lethal. The classical approach to avoiding it is to cool cells slowly enough so that nearly all their supercooled freezable water leaves the cell osmotically before they have cooled to a temperature that permits IIF. An alternative approach is to cool the cell rapidly to just above its ice nucleation temperature, and hold it there long enough to permit dehydration. Then, the cell is cooled rapidly to -70 degrees C or below. This approach, often called interrupted rapid cooling, is the subject of this paper. Mouse oocytes were suspended in 1.5M ethylene glycol (EG)/PBS, rapidly cooled (50 degrees C/min) to -25 degrees C and held for 5, 10, 20, 30, or 40 min before being rapidly cooled (50 degrees C/min) to -70 degrees C. In cells held for 5 min, IIF (flashing) occurred abruptly during the second rapid cool. As the holding period was increased to 10 and 20 min, fewer cells flashed during the cooling and more turned black during warming. Finally, when the oocytes were held 30 or 40 min, relatively few flashed during either cooling or warming. Immediately upon thawing, these oocytes were highly shrunken and crenated. However, upon warming to 20 degrees C, they regained most of their normal volume, shape, and appearance. These oocytes have intact cell membranes, and we refer to them as survivors. We conclude that 30 min at -25 degrees C removes nearly all intracellular freezable water, the consequence of which is that IIF occurs neither during the subsequent rapid cooling to -70 degrees C nor during warming.
Peter Mazur; Irina L Pinn; F W Kleinhans
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2007-07-04
Journal Detail:
Title:  Cryobiology     Volume:  55     ISSN:  0011-2240     ISO Abbreviation:  Cryobiology     Publication Date:  2007 Oct 
Date Detail:
Created Date:  2007-09-03     Completed Date:  2008-01-09     Revised Date:  2014-04-08    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  158-66     Citation Subset:  IM    
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MeSH Terms
Cryopreservation / methods*
Mice, Inbred ICR
Oocytes / chemistry*
Time Factors
Transition Temperature
Water / chemistry*
Grant Support
R01 RR018470/RR/NCRR NIH HHS; R01 RR018470-04/RR/NCRR NIH HHS; R01 RR18470/RR/NCRR NIH HHS
Reg. No./Substance:
Comment In:
Cryobiology. 2008 Feb;56(1):22-7   [PMID:  18045584 ]

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