Abstract
To investigate the effect of calcium on the proliferation and differentiation of murine corneal epithelial cells in vitro. Mouse corneal epithelial cells were cultured in serum-free low-Ca(2+) medium (KSFM) and KSFM supplemented with 0.9mmol/L Ca(2+). Population doublings (PDs) were determined. The expression of corneal epithelial cell markers p63, keratin 19 (K19) and involucrin was investigated by RT-PCR analysis and semiquantitative analysis of Western blotting. Cells in KSFM were stably subcultured over 25 passages, however, none of the cell lines could pass P4 in KSFM with Ca(2+). In KSFM, the cells was were homogeneous and small cells with typical cobblestone appearance; and expressed p63, K19 and involucrin. After medium was supplemented with calcium, cells became a heterogeneous mix of small and large cells. Furthermore, semiquantitative analysis of Western blotting showed that the expression of involucrin was increased significantly. Calcium has the effect of inhibiting proliferation and triggering differentiation on mouse corneal epithelial cells.
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