Document Detail


Cryopreservation of adherent human embryonic stem cells.
MedLine Citation:
PMID:  15486934     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Standard human embryonic stem (HES) cell cryopreservation methodologies, including slow freezing and vitrification of colonies in suspension, are plagued by poor viability and high differentiation rates upon recovery. To facilitate research studies and clinical applications of HES cells, we have developed a cryopreservation technique based on stabilizing HES colonies adherent to or embedded in a Matrigel matrix. This method increases cell viability by over an order of magnitude compared with cryopreservation in suspension and reduces differentiation. Loading adherent HES cells with the disaccharide trehalose prior to cryopreserving in a dimethylsulfoxide-containing cryoprotectant solution further improves cell viability under certain conditions. Our proposed approach has the potential to reduce the time required to amplify frozen stocks of HES cells, minimize risk of clonal selection during freeze-thaw cycles, and facilitate storage of HES cell clone libraries.
Authors:
Lin Ji; Juan J de Pablo; Sean P Palecek
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  88     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2004-10-25     Completed Date:  2005-03-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  299-312     Citation Subset:  IM    
Copyright Information:
Copyright 2004 Wiley Periodicals, Inc.
Affiliation:
Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, Wisconsin 53706, USA.
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MeSH Terms
Descriptor/Qualifier:
Biocompatible Materials / chemistry
Cell Adhesion / physiology*
Cell Culture Techniques / methods*
Cell Line
Cell Survival / physiology
Collagen / chemistry*
Cryopreservation / methods*
Dose-Response Relationship, Drug
Drug Combinations
Humans
Laminin / chemistry*
Proteoglycans / chemistry*
Stem Cells / chemistry,  cytology*,  drug effects,  physiology*
Tissue Engineering / methods*
Trehalose / chemistry,  pharmacology*
Chemical
Reg. No./Substance:
0/Biocompatible Materials; 0/Drug Combinations; 0/Laminin; 0/Proteoglycans; 119978-18-6/matrigel; 9007-34-5/Collagen; 99-20-7/Trehalose

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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