Document Detail


Isolation and cultivation of porcine astrocytes.
MedLine Citation:
PMID:  22144305     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
A procedure for the isolation and cultivation of astrocytes from swine is described. More specifically, the donor animals are adolescent minipigs about 3 months in age and 10 kg in weight. About 20 g of cerebral tissue can be isolated from the piglet, yielding enough astrocytes of homogeneous genetics for experimentation after only one passage in culture. The astrocyte isolation procedure includes mechanical and enzymatic digestion of the brain tissue followed by separation of the brain fragments, based on size and density. Astrocytes are further purified from any residual nonastrocytes by differential attachment during the first passage. The resulting culture is purely astrocytes (>98%) based upon their appearance in phase-contrast microscopy and their uniform expression of glial fibrillary acidic protein. More importantly for our purposes, the astrocytes greatly enhanced the functionality of our in vitro blood-brain barrier model when cocultured with porcine brain capillary endothelial cells.
Authors:
Dennis J Bobilya
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  814     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2012  
Date Detail:
Created Date:  2011-12-06     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  127-35     Citation Subset:  IM    
Affiliation:
Department of Molecular, Cellular and Biomedical Sciences, University of New Hampshire, Durham, NH, USA, dbobilya@unh.edu.
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