| MK/T-1, an immortalized fibroblast cell line derived using cultures of mouse corneal stroma. | |
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MedLine Citation:
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PMID: 11344338 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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R L Gendron; C Y Liu; H Paradis; L C Adams; W W Kao |
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Publication Detail:
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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:
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Title: Molecular vision Volume: 7 ISSN: 1090-0535 ISO Abbreviation: Mol. Vis. Publication Date: 2001 May |
Date Detail:
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Created Date: 2001-05-09 Completed Date: 2001-08-23 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 9605351 Medline TA: Mol Vis Country: United States |
Other Details:
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Languages: eng Pagination: 107-13 Citation Subset: IM |
Affiliation:
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Department of Pediatrics, Division of Hematology and Oncology, Children's Hospital Medical Center, University of Cincinnati, Cincinnati, OH 45229-3039, USA. rlgendron@chmcc.org |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals 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 Mice, Inbred C57BL Mitogen-Activated Protein Kinases / metabolism Proteoglycans / metabolism RNA* Telomerase / genetics, metabolism Transfection Transforming Growth Factor beta / pharmacology p38 Mitogen-Activated Protein Kinases |
| Grant Support | |
ID/Acronym/Agency:
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EY10556/EY/NEI NIH HHS; EY11845/EY/NEI NIH HHS; EY12486/EY/NEI NIH HHS; EY12827/EY/NEI NIH HHS; U10CA13539/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cytoskeletal Proteins; 0/DNA-Binding Proteins; 0/Proteoglycans; 0/Transforming Growth Factor beta; 0/telomerase RNA; 63231-63-0/RNA; EC 2.7.11.24/Mitogen-Activated Protein Kinases; EC 2.7.11.24/p38 Mitogen-Activated Protein Kinases; EC 2.7.7.49/Telomerase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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