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K. Izumi, D. Kurosaka, T. Iwata, K. Nakamura, Y. Ohashi, Y. Oguchi, Y. Tanaka, K. Tsubota, Y. Mashima; Corneal Epithelial Cells Are Essential for Prevention of Myodifferentiation of Corneal Fibroblasts in a Coculture System . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2155.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To establish a human corneal epithelial cell–fibroblast coculture system and to demonstrate corneal epithelial–mesenchymal interactions and the influence of growth factors on myofibroblast differentiation of corneal fibroblast using our coculture model. Methods: Corneal fibroblasts were cultured on collagen gel with or without confluent corneal epithelial cells. To evaluate myodifferentiation of fibroblasts, collagen gel contraction was measured over 6 days with or without antibody against transforming growth factor (TGF)–ß. Amounts of mRNA encoding TGF–ß2 and α–smooth muscle actin (SMA) were determined by real–time quantitative PCR. Procedures were repeated with cell exposure to antibody against basic fibroblast growth factor (bFGF) antibody. Basic FGF was assayed in media on days 3 and 6. To evaluate the nature of fibroblast interaction with epithelial cells, fibroblasts cultured in conditioned medium collected from epithelial cells were subjected to gel contraction measurement and real–time quantitative PCR. Results: Collagen gel contraction was increased on gels with fibroblasts alone, but suppressed when fibroblasts were cocultured with epithelial cells. Anti–TGF–ß antibody eliminated gel contraction almost completely. TGF–ß2 and α–SMA mRNA expression increased with time in fibroblasts cultured alone, but changed little in cocultured fibroblasts. Coculture media contained significant amounts of bFGF. Anti–bFGF antibody increased gel contraction of cocultured fibroblasts, and expression of TGF–ß2 and α–SMA mRNA. Conditioned medium suppressed gel contraction and α–SMA mRNA expression in fibroblasts, but less than coculture. Conclusions: Our coculture model is useful for the investigations of corneal epithelial cell–fibroblast interaction. Corneal epithelial cells suppress differentiation of corneal fibroblasts into myofibroblasts and prevent corneal stromal scarring via soluble factors including bFGF.
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