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Cryopreservation of human embryonic stem cells without the use of a programmable freezer

DC Field Value Language
dc.contributor.authorHa, Sung Yun-
dc.contributor.authorJee, Byung Chul-
dc.contributor.authorSuh, Chang Suk-
dc.contributor.authorKim, Hee Sun-
dc.contributor.authorOh, Sun Kyung-
dc.contributor.authorKim, Seok Hyun-
dc.contributor.authorMoon, Shin Yong-
dc.date.accessioned2009-11-04-
dc.date.available2009-11-04-
dc.date.issued2005-
dc.identifier.citationHum Reprod 20,1779-1785en
dc.identifier.issn0268-1161 (Print)-
dc.identifier.urihttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15760949-
dc.identifier.urihttps://hdl.handle.net/10371/11068-
dc.description.abstractBACKGROUND: An effective freezing-thawing technique is crucial for the clinical application of human embryonic stem (ES) cells. The aim of this study was to find an optimal cryopreservation protocol for human ES cells using slow freezing-rapid thawing without a programmable freezer. METHODS: The human ES cell line, SNUhES-3, was cultured on an STO feeder layer in gelatin-coated tissue culture dishes. All cryopreservation steps were performed using a simple commercial freezing container. The survival rate of cryopreserved-thawed human ES cells was estimated by counting colony numbers under a stereomicroscope. Initially, we compared the survival rates of cryopreserved human ES cells using three cryoprotectants: dimethylsulphoxide (DMSO), ethylene glycol (EG) and glycerol. In this experiment, 5% DMSO/95% fetal bovine serum (FBS) (vol/vol) showed the highest survival rate. We next tested the impact of various concentrations of FBS (95, 50 and 5%) with 5% DMSO, and then examined the effects of adding EG or glycerol to 5% DMSO + optimal FBS. RESULTS: No significant difference in survival rate was observed between 95 and 50% FBS in the presence of 5% DMSO. A significant improvement in survival rate was obtained by adding 10% EG to 5% DMSO+50% FBS. After thawing, surviving cells were found to maintain the inherent characteristics of human ES cells. CONCLUSION: 5% DMSO+50% FBS+10% EG may be an optimal cryoprotectant for the slow freezing-rapid thawing of human ES cells.en
dc.language.isoen-
dc.publisherOxford University Pressen
dc.subjectcryopreservationen
dc.subjectdimethylsulphoxideen
dc.subjectethylene glycolen
dc.subjecthuman embryonic stem cellsen
dc.titleCryopreservation of human embryonic stem cells without the use of a programmable freezeren
dc.typeArticleen
dc.contributor.AlternativeAuthor지병철-
dc.contributor.AlternativeAuthor서창석-
dc.contributor.AlternativeAuthor김희선-
dc.contributor.AlternativeAuthor오선경-
dc.contributor.AlternativeAuthor김석현-
dc.contributor.AlternativeAuthor문신용-
dc.contributor.AlternativeAuthor하성윤-
dc.identifier.doi10.1093/humrep/deh854-
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