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AIM: To investigate the effect of calcium on the proliferation and differentiation of murine corneal epithelial cells in vitro.METHODS: Mouse corneal epithelial cells were cultured in serum-free low-Ca2+ medium (KSFM) and KSFM supplemented with 0.9mmol/L Ca2+.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.RESULTS: Cells in KSFM were stably subcultured over 25 passages,however,none of the cell lines could pass P4 in KSFM with Ca2+.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.CONCLUSION: Calcium has the effect of inhibiting proliferation and triggering differentiation on mouse corneal epithelial cells.
AIM: To investigate the effect of calcium on the proliferation and differentiation of murine corneal epithelial cells in vitro. METHODS: Mouse corneal epithelial cells were cultured in serum-free low-Ca2 + medium (KSFM) and KSFM supplemented with 0.9 mmol / L Ca2 +. 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 .RESULTS: Cells in KSFM were stably subcultured over 25 passages, However, none of the cell lines could pass P4 in KSFM with Ca2 +. 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. Still further, semiquantitative analysis of Western blotting showed that the expression of involucrin was increased significantly. CONCLUSION: Calcium has the effect of inhi biting proliferation and triggering differentiation on mouse corneal epithelial cells.