Comparison of three methods for cryopreservation of human embryonic stem cells.
OBJECTIVE
To establish reliable methods for cryopreservation of human embryonic stem cells (hESCs).


METHODS
Prospective experimental study.


METHODS
University laboratory.


METHODS
One hESC line.


METHODS
The attachment rates and recovery rates of cryopreserved hESCs using three different cryopreservation methods were compared.


METHODS
The hESCs were frozen and thawed by conventional cryopreservation, programmable cryopreservation, and vitrification method. The efficiency of cryopreservation was assessed by attachment rate and recovery rate.


RESULTS
The attachment rate and recovery rate after thawing of hESCs frozen by the conventional cryopreservation method were significantly lower than those of hESCs frozen by programmed cryopreservation and vitrification methods. Vitrification resulted in the highest attachment rate and recovery rate compared with the other two methods. Human ESCs after vitrification and programmable cryopreservation still expressed pluripotent markers, maintained normal karyotype, and retained their pluripotency.


CONCLUSIONS
Our data show that programmable cryopreservation and vitrification methods are appropriate for cryopreservation of hESCs, whereas the conventional slow-rate freezing method is not appropriate for cryopreservation of hESCs.
Li
Yang
Y
Stem Cell Research Center and Department of Cell Biology, Peking University Health Science Center, Beijing 100083, People's Republic of China.
Tan
Ji-Chun
JC
Li
Ling-Song
LS
D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D015925:Cryopreservation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007621:Karyotyping; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D013696:Temperature; D013724:Teratoma / Q000473:pathology
D015415:Biomarkers