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M.M. Jumblatt, C.G. Emberts, Y. Li, P.A. Overbeek, J.E. Jumblatt; Conjunctival Differentiation: Role of FGF-7 and Extracellular Matrix . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3782.
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Purpose: The conjunctival epithelium contains both squamous and goblet cells. In mice that over-express FGF-7, goblet-like cells also develop at the corneal surface. The purpose of this study was to examine the corneas of normal and transgenic mice for goblet cell markers and to determine if FGF-7 induced goblet cell differentiation in cultured human conjunctival epithelium. Methods: Corneal and conjunctival tissues were collected from transgenic mice over-expressing lenticular KGF-7. These tissues were analyzed for the expression of the goblet cell mucin MUC5AC by RT-PCR. Human conjunctival epithelial cells were isolated and propagated in defined medium. Cells were sub-cultured in the absence or presence of FGF-7 and ECM for 10 days. Cultures were either fixed for histological examination or analyzed for the presence of FGF-7, its specific receptor (KGFR2) by RT-PCR. Results: MurineMUC5AC mRNA was expressed in the conjunctival tissue of normal and transgenic mice. MUC5AC was expressed ectopically in the corneas of transgenic, but not wild type mice. Human conjunctival fibroblasts expressed FGF-7 mRNA but not that of KGFR2. Human conjunctival epithelial cells grown on ECM in the presence of FGF-7 formed lobed invaginations, which in cross-section appeared as tubules and spheres. These cultured cells expressed KGFR2, but not FGF-7. Despite their differentiated appearance, human conjunctival epithelial cells cultured in the presence of FGF-7 and ECM did not express MUC5AC mRNA. Conclusions: MUC5AC is expressed ectopically in FGF-7 transgenic mice. FGF-7 and/or ECM enhance conjunctival epithelial cell growth in vitro, but are insufficient to promote goblet cell differentiation as defined by expression of MUC5AC.
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