Document Detail


Rapid purification of transfected porcine muscle cells.
MedLine Citation:
PMID:  11264956     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A detailed methodology is described for fluorescence-activated cell sorting (FACS) of porcine muscle cells that have been transfected to express green fluorescent protein (GFP). Cells are liberated from porcine skeletal muscle and primary cultures are transfected with DNA encoding GFP. Primary cultures are subjected to immunocytochemistry using a primary muscle-specific monoclonal antibody followed by incubation with a phycoerythrin-conjugated second antibody. Transfected myoblasts are sorted from fibroblasts using forward angle light scatter and ninety degree light scatter, phycoerythrin fluorescence, and GFP fluorescence. These procedures allow for isolation of genetically- engineered porcine muscle cells more rapidly than traditional clonal selection procedures. Consequently, FACS provides porcine myoblast populations that retain the majority of their replicative capacity and are not contaminated with non-myogenic cells.
Authors:
J R Blanton; J P Robinson; D E Gerrard; C A Bidwell; A L Grant
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Methods in cell science : an official journal of the Society for In Vitro Biology     Volume:  22     ISSN:  1381-5741     ISO Abbreviation:  Methods Cell Sci     Publication Date:  2000  
Date Detail:
Created Date:  2001-03-26     Completed Date:  2001-05-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9515839     Medline TA:  Methods Cell Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  217-23     Citation Subset:  IM    
Affiliation:
Department of Animal Science and Food Technology, Texas Tech University, Lubbock, TX, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal
Cell Separation
Flow Cytometry / methods*
Gene Transfer Techniques
Green Fluorescent Proteins
Immunohistochemistry
Luminescent Proteins / genetics*
Muscle, Skeletal / cytology*
Phycoerythrin / immunology
Swine
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Luminescent Proteins; 11016-17-4/Phycoerythrin; 147336-22-9/Green Fluorescent Proteins

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


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