Document Detail

MK/T-1, an immortalized fibroblast cell line derived using cultures of mouse corneal stroma.
MedLine Citation:
PMID:  11344338     Owner:  NLM     Status:  MEDLINE    
PURPOSE: Immortalized cell lines representing fibroblast cells from corneal stroma would facilitate studies of corneal cell biology and injury response. METHODS: Primary cultures of cells derived from mouse corneal stroma were transfected with a human telomerase reverse transcriptase (hTERT) expression construct to maximize chances of cellular immortalization. A resulting cell line was analyzed for telomerase activity, cell growth characteristics, senescence and gene expression patterns. Specific responses to transforming growth factor beta (TGF-beta) were also analyzed. RESULTS: An immortalized cell line was derived and was named MK/T-1. MK/T-1 cells show no signs of cellular senescence or transformation at over 100 passages. Telomerase activity was significantly higher in MK/T-1 cells as compared to the parental cell cultures. However, relative telomere length (RTL) in the MK/T-1 and parental cells was not significantly different. Senescence associated beta-galactosidase (SA-beta-Gal) activity was not detected in late passage MK/T-1 cells while the parental cells had already upregulated SA-beta-Gal at high levels by passage 9. The MK/T-1 cells express vimentin, tubulin, lumican, mimecan, decorin and collagen I, but not keratocan. Exposure of the MK/T-1 cells to TGF-beta induces the expression of smooth muscle alpha-actin (ASMA), the activation of MAP Kinase (p38-MAPK) and morphological changes consistent with cytoskeletal reorganization. CONCLUSIONS: MK/T-1 cells represent an immortalized fibroblast cell line derived using cultures from corneal stroma cell preparations. Expression of hTERT may contribute to immortalization of the MK/T-1 cells by a mechanism other than increases in RTL. MK/T-1 cells may be a useful model in which to study the responses of corneal fibroblast cells to cytokines and other diverse environmental factors in vitro.
R L Gendron; C Y Liu; H Paradis; L C Adams; W W Kao
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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.     Date:  2001-05-08
Journal Detail:
Title:  Molecular vision     Volume:  7     ISSN:  1090-0535     ISO Abbreviation:  Mol. Vis.     Publication Date:  2001 May 
Date Detail:
Created Date:  2001-05-09     Completed Date:  2001-08-23     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9605351     Medline TA:  Mol Vis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  107-13     Citation Subset:  IM    
Department of Pediatrics, Division of Hematology and Oncology, Children's Hospital Medical Center, University of Cincinnati, Cincinnati, OH 45229-3039, USA.
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MeSH Terms
Blotting, Northern
Blotting, Western
Cell Aging
Cell Line, Transformed
Corneal Stroma / cytology*,  metabolism
Cytoskeletal Proteins / metabolism
Cytoskeleton / metabolism
DNA-Binding Proteins
Fibroblasts / cytology*,  metabolism
Mice, Inbred C57BL
Mitogen-Activated Protein Kinases / metabolism
Proteoglycans / metabolism
Telomerase / genetics,  metabolism
Transforming Growth Factor beta / pharmacology
p38 Mitogen-Activated Protein Kinases
Grant Support
Reg. No./Substance:
0/Cytoskeletal Proteins; 0/DNA-Binding Proteins; 0/Proteoglycans; 0/Transforming Growth Factor beta; 0/telomerase RNA; 63231-63-0/RNA; EC Protein Kinases; EC Mitogen-Activated Protein Kinases; EC

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