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Y. Pei, D.M. Sherry, A.M. McDermott; Thy-1, a Marker for Corneal Fibroblasts . Invest. Ophthalmol. Vis. Sci. 2003;44(13):869.
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Purpose: After corneal injury, the normal corneal keratocyte becomes activated and transforms into a repair phenotype, the corneal fibroblast. Thy-1, a cell surface protein, can be used to distinguish a subpopulation of fibroblasts in some tissues, and is highly expressed by fibroblasts during repair. In the present study we have investigated if Thy-1 is expressed by corneal keratocytes and the corneal fibroblast phenotype to determine if Thy-1 can be used as a marker to distinguish between these two cell types. Methods: Human corneal keratocytes were isolated by collagenase digestion. The cells were either processed immediately (i.e. freshly isolated keratocytes) or were cultured in the presence of 10% fetal bovine serum to induce transformation to the corneal fibroblast phenotype. Thy-1 mRNA and protein expression by freshly isolated keratocytes and corneal fibroblasts (passages 0 to 4) were assessed by RT-PCR and Western blotting. RNA was also extracted from the whole stroma and assessed by RT-PCR. Immunolabeling was performed to localize Thy-1 in cultured human corneal fibroblasts and in normal human corneal tissue sections. Results: Thy-1 mRNA and protein were detectable in cultured human corneal fibroblasts (n=2-3), but not freshly isolated keratocytes (n=2-3). Thy-1 mRNA was not detected in extracts of whole stroma. Early passages of corneal fibroblasts showed low Thy-1 protein expression, which increased with passaging. Cultured human corneal fibroblasts (n=2) showed positive labeling for Thy-1, but no detectable Thy-1 staining was found in the keratocytes in the stroma of normal human corneal tissue sections (n=2). Conclusions: Corneal fibroblasts but not keratocytes express Thy-1 mRNA and protein. This suggests that Thy-1 can be used as a marker to distinguish the normal keratocyte from its repair phenotype, the corneal fibroblast.
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