Document Detail


Isolation and characterization of porcine Müller cells. Myofibroblastic dedifferentiation in culture.
MedLine Citation:
PMID:  8603859     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To characterize phenotypic and antigenic changes in isolated Müller cells during proliferation in extended culture. Methods: Müller cells were isolated from porcine retina by sequential papain and DNase digestion, trituration, and density gradient centrifugation. The identity of the isolated cells was confirmed by immunodetection of carbonic anhydrase II (CA-II), cellular retinaldehyde-binding protein (CRALBP), glial fibrillary acidic protein (GFAP), vimentin, and delta smooth muscle actin (alpha SMA). Continuously proliferating cells established in culture were examined for changes in the expression of these antigens. RESULTS: Primary cultures of purified Müller cells, incubated under routine culture conditions, were proliferative and lost immunodetectable CRALBP within 2 weeks. The expression of CA-II also diminished with time, but at an apparently lower rate than that of CRALBP. Loss of GFAP expression was even more gradual and was complete by passage 5. De novo expression of alpha SMA was detectable in some cells within 12 days in culture and by all cells by passage 5. During this period, vimentin expression remained qualitatively unchanged. CONCLUSIONS: Isolated porcine Muller cells in culture undergo a phenotypic dedifferentiation to a fibroblast-like cell, which includes loss of detectable CRALBP, CA-II, and GFAP, and they acquire expression of the myoid marker alpha SMA.
Authors:
C Guidry
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  37     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  1996 Apr 
Date Detail:
Created Date:  1996-05-16     Completed Date:  1996-05-16     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  740-52     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, Eye Foundation Hospital, University of Alabama at Birmingham, 35294, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Carbonic Anhydrases / metabolism
Carrier Proteins / metabolism
Cell Differentiation
Cell Division
Cell Separation
Cells, Cultured
Cytoskeletal Proteins / metabolism
Electrophoresis, Polyacrylamide Gel
Fibroblasts / cytology*,  metabolism
Fluorescent Antibody Technique
Glial Fibrillary Acidic Protein / metabolism
Neuroglia / cytology*,  metabolism
Phenotype
Retinaldehyde / metabolism
Swine
Grant Support
ID/Acronym/Agency:
EY09536/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/11-cis-retinal-binding protein; 0/Carrier Proteins; 0/Cytoskeletal Proteins; 0/Glial Fibrillary Acidic Protein; 116-31-4/Retinaldehyde; EC 4.2.1.1/Carbonic Anhydrases

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


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