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Title: Low temperature tolerance of human embryonic stem cells
Authors: Boon, C.H. 
Vinoth, K.J. 
Liu, H. 
Hande, M.P.
Cao, T. 
Keywords: Human embryonic
Low temperature
Stem cells
Issue Date: 2006
Citation: Boon, C.H.,Vinoth, K.J.,Liu, H.,Hande, M.P.,Cao, T. (2006). Low temperature tolerance of human embryonic stem cells. International Journal of Medical Sciences 3 (4) : 124-129. ScholarBank@NUS Repository.
Abstract: This study investigated the effects of exposing human embryonic stem cells (hESC) to 4°C and 25°C for extended durations of 24h and 48h respectively. Cell survivability after low temperature exposure was assessed through the MTT assay. The results showed that hESC survivability after exposure to 25°C and 4°C for 24h was 77.3 ± 4.8 % and 64.4 ± 4.4 % respectively (significantly different, P < 0.05). The corresponding survival rates after 48h exposure to 25°C and 4°C was 71.0 ± 0.5 % and 69.0 ± 2.3 % respectively (not significantly different, P > 0.05). Spontaneous differentiation of hESC after low temperature exposure was assessed by morphological observations under bright-field and phase-contrast microscopy, and by immunocytochemical staining for the pluripotency markers SSEA-3 and TRA-1-81. hESC colonies were assigned into 3 grades according to their degree of spontaneous differentiation: (1) Grade A which was completely or mostly undifferentiated, (2) Grade B which was partially differentiated, and (3) Grade C which was mostly differentiated. In all low temperature exposed groups, about 95% of colonies remain undifferentiated (Grade A), which was not significantly different (P > 0.05) from the unexposed control group maintained at 37°C. Additionally, normal karyotype was maintained in all low temperature-exposed groups, as assessed by fluorescence in situ hybridization (FISH) of metaphase spreads with telomere and centromere-specific PNA probes. Further analysis with m-FISH showed that chromosomal translocations were absent in all experimental groups. Hence, hESC possess relatively high-tolerance to extended durations of low temperature exposure, which could have useful implications for the salvage of hESC culture during infrequent occurrences of incubator break-down and power failure. ©2006 Ivyspring International Publisher. All rights reserved.
Source Title: International Journal of Medical Sciences
ISSN: 14491907
Appears in Collections:Staff Publications

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