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. 2011;4(3):247-9.
doi: 10.3980/j.issn.2222-3959.2011.03.06. Epub 2011 Jun 18.

Effect of calcium on the proliferation and differentiation of murine corneal epithelial cells in vitro

Xiao-Li Ma  1 , Han-Qiang Liu
Affiliations

Effect of calcium on the proliferation and differentiation of murine corneal epithelial cells in vitro

Xiao-Li Ma et al. Int J Ophthalmol. 2011.

Abstract

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-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.

Results: 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.

Conclusion: Calcium has the effect of inhibiting proliferation and triggering differentiation on mouse corneal epithelial cells.

Keywords: calcium; cell culture; cornea; epithelium.

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Figures

Figure 1
Figure 1. Population doubling (PD) of cells cultured in KSFM and KSFM+Ca
Figure 2
Figure 2. Morphology of cells cultured in
A: KSFM; B: KSFM with high Ca2+
Figure 3
Figure 3. p63, K19 and IVL expression in different medium
A: RT-PCR; B: Western blotting
See this image and copyright information in PMC

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