Document Detail

Kinetics of cell death of frozen-thawed human embryonic stem cell colonies is reversibly slowed down by exposure to low temperature.
MedLine Citation:
PMID:  17266792     Owner:  NLM     Status:  MEDLINE    
A major challenge in the widespread application of hES (human embryonic stem) cells in clinical therapy and basic scientific research is the development of efficient cryopreservation protocols. Conventional slow-cooling protocols utilizing standard cryoprotectant concentrations i.e. 10% (v/v) DMSO, yield extremely low survival rates of less than 5% as reported by previous studies. This study characterized cell death in frozen-thawed hES colonies that were cryopreserved under standard conditions. Surprisingly, our results showed that immediately after post-thaw washing, the overwhelming majority of hES cells were viable (approximately 98%), as assessed by the trypan blue exclusion test. However, when the freshly thawed hES colonies were placed in a 37 degrees C incubator, there was a gradual reduction in cell viability over time. The kinetics of cell death was drastically slowed down by keeping the freshly thawed hES colonies at 4 degrees C, with more than 90% of cells remaining viable after 90 min of incubation at 4 degrees C. This effect was reversible upon re-exposing the cells to physiological temperatures. The vast majority of low temperature-exposed hES colonies gradually underwent cell death upon incubation for a further 90 min at 37 degrees C. Hence, our observations would strongly suggest involvement of a self-induced apoptotic mechanism, as opposed to cellular necrosis arising from cryoinjury.
B C Heng; C P Ye; H Liu; W S Toh; A J Rufaihah; T Cao
Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  Zygote (Cambridge, England)     Volume:  14     ISSN:  0967-1994     ISO Abbreviation:  Zygote     Publication Date:  2006 Nov 
Date Detail:
Created Date:  2007-02-01     Completed Date:  2007-03-07     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9309124     Medline TA:  Zygote     Country:  England    
Other Details:
Languages:  eng     Pagination:  341-8     Citation Subset:  IM    
National University of Singapore, Singapore.
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MeSH Terms
Cell Adhesion
Cell Death
Cell Survival
Cells, Cultured
Cold Temperature
Cryopreservation / methods*
Embryonic Stem Cells* / cytology

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