Journal
FERTILITY AND STERILITY
Volume 93, Issue 3, Pages 999-1005Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.fertnstert.2008.10.052
Keywords
Human embryonic stem cell; cryopreservation methods; cryopreservation efficiency
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Funding
- Chinese National 973 Project [2006CB943603, 2006CB503905]
- Chinese National 863 Project [2006AA02A114]
- Natural Science foundation of the Beijing Ministry of Science and Technology [D0750701350704]
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Objective: To establish reliable methods for cryopreservation of human embryonic stem cells (hESCs). Design: Prospective experimental study. Setting: University laboratory. Patient(s): One hESC line. Intervention(S): The attachment rates and recovery rates of cryopreserved hESCs using three different cryopreservation methods were compared. Main Outcome Measure(s): 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. Result(S): 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. Conclusion(s): Our data show that program in able cryopreservation and vitrification methods are appropriate for cryopreservation of hESCs, whereas the conventional slow-rate freezing method is not appropriate for cryopreservation of hESCs. (Fertil Steril (R) 2010;93:999-1005. (C)2010 by American Society for Reproductive Medicine.)
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