Document Detail


Integrated culture and purification of rat Schwann cells from freshly isolated adult tissue.
MedLine Citation:
PMID:  23060244     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe a simple, rapid and highly selective protocol for the primary culture of Schwann cells in vitro from freshly dissociated adult rat nerve. The protocol is based on a selective culture medium comprising both mitogens (forskolin and optionally N2 supplement plus bovine pituitary extract), to stimulate growth of Schwann cells, plus an inhibitory substrate to simultaneously restrict fibroblast overgrowth (D-valine), contained in DMEM. This protocol differs from other available methods in that it uses the preferential capacity of Schwann cells to metabolize D-valine because of the difference in expression of a D-amino acid oxidase (DAAO) enzyme between Schwann cells and fibroblasts plus the presence of a selective mitogen to stimulate growth of Schwann cells. This permits derivation of highly pure Schwann cells directly from fresh adult nerve. Average Schwann cell purities of 97% can be achieved after 19 d without pre-degeneration, purification or antimitotic steps.
Authors:
Rossukon Kaewkhaw; Andy M Scutt; John W Haycock
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-10-11
Journal Detail:
Title:  Nature protocols     Volume:  7     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-10-12     Completed Date:  2013-03-19     Revised Date:  2013-03-21    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1996-2004     Citation Subset:  IM    
Affiliation:
Department of Materials Science & Engineering, Kroto Research Institute, University of Sheffield, Sheffield, UK.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques*
Culture Media
Male
Rats
Rats, Wistar
Schwann Cells / cytology*,  metabolism
Sciatic Nerve / cytology
Valine / metabolism
Chemical
Reg. No./Substance:
0/Culture Media; 7004-03-7/Valine

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


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