Please use this identifier to cite or link to this item: https://doi.org/10.1017/S0967199406003893
Title: Kinetics of cell death of frozen-thawed human embryonic stem cell colonies is reversibly slowed down by exposure to low temperature
Authors: Heng, B.C. 
Ye, C.P.
Liu, H. 
Toh, W.S. 
Rufaihah, A.J. 
Cao, T. 
Keywords: Cell death
Cryopreservation
Embryonic
Human
Stem cells
Issue Date: Nov-2006
Source: Heng, B.C., Ye, C.P., Liu, H., Toh, W.S., Rufaihah, A.J., Cao, T. (2006-11). Kinetics of cell death of frozen-thawed human embryonic stem cell colonies is reversibly slowed down by exposure to low temperature. Zygote 14 (4) : 341-348. ScholarBank@NUS Repository. https://doi.org/10.1017/S0967199406003893
Abstract: 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°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°C, with more than 90% of cells remaining viable after 90 min of incubation at 4°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°C. Hence, our observations would strongly suggest involvement of a self-induced apoptotic mechanism, as opposed to cellular necrosis arising from cryoinjury. © 2006 Cambridge University Press.
Source Title: Zygote
URI: http://scholarbank.nus.edu.sg/handle/10635/52533
ISSN: 09671994
DOI: 10.1017/S0967199406003893
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