Document Detail

Cryopreservation of primate embryonic stem cells with chemically-defined solution without Me2SO.
MedLine Citation:
PMID:  19857481     Owner:  NLM     Status:  MEDLINE    
Human embryonic stem (hES) cells are expected to be useful in the fields of regenerative medicine and tissue engineering due to their pluripotency. Therefore, it is necessary to establish highly efficient and reliable methods for the cryopreservation of hES cells. We have cryopreserved cynomolgus and human ES cells by the vitrification method, using a chemically-defined dimethyl sulfoxide (Me(2)SO)-free and serum-free medium composed of Euro-Collins solution as a base medium and 40% (v/v) ethylene glycol (EG) and 10% (w/v) polyethylene glycol (PEG) as cryoprotectants. When the vitrification and the cryoprotectants were combined, the recovery ratio of hES cells was 22.9+/-7.7%, compared to 0.4+/-0.2% when the conventional slow-freezing method was used. After the cryopreservation and thawing cycle, hES cells were easily cultured and expressed undifferentiated cell markers such as Nanog, Oct-4, SSEA-4, and alkaline phosphatase activity after several subculturing steps. We also found that the pluripotency of hES cells was maintained, as demonstrated by teratoma formation of ES cells transplanted into severe combined immunodeficient (SCID) mice. Thus, we conclude that we have successfully cryopreserved primate ES cells with high efficiency using a Me(2)SO-free, chemically-defined medium.
Tatsuya Nishigaki; Yuji Teramura; Hirofumi Suemori; Hiroo Iwata
Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-10-24
Journal Detail:
Title:  Cryobiology     Volume:  60     ISSN:  1090-2392     ISO Abbreviation:  Cryobiology     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-03-08     Completed Date:  2010-05-25     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  159-64     Citation Subset:  IM    
Copyright Information:
Copyright 2009 Elsevier Inc. All rights reserved.
Department of Reparative Materials, Institute for Frontier Medical Sciences, Kyoto University, Japan.
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MeSH Terms
Biological Markers / metabolism
Cell Differentiation
Cryopreservation / methods*
Cryoprotective Agents
Dimethyl Sulfoxide
Embryonic Stem Cells* / cytology,  metabolism,  transplantation
Ethylene Glycol
Macaca fascicularis
Mice, SCID
Pluripotent Stem Cells / cytology,  metabolism,  transplantation
Polyethylene Glycols
Teratoma / etiology,  pathology
Reg. No./Substance:
0/Biological Markers; 0/Cryoprotective Agents; 0/Polyethylene Glycols; 0/Solutions; 107-21-1/Ethylene Glycol; 67-68-5/Dimethyl Sulfoxide

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