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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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References

    1. Ang LP, Tan DT. Ocular surface stem cells and disease: Current concepts and clinical applications. Ann Acad Med Singapore. 2004;33(5):576–580. - PubMed
    1. Hiraki A, Shirasuna K, Ikari T, Shinohara M, Garrod DR. Calcium induces differentiation of primary human salivary acinar cells. J Cell Physiol. 2002;193(1):55–63. - PubMed
    1. D'Souza SJ, Pajak A, Balazsi K, Dagnino L. Ca2+ and bmp-6 signaling regulate e2f during epidermal keratinocyte differentiation. J Biol Chem. 2001;276(26):23531–23538. - PubMed
    1. Ma XL, Kong J, Liu HQ, Zhao JY, Zhang JS. Comparative study of four culture methods to engineer murine corneal epithelial sheet. Guoji Yanke Zazhi(Int J Ophthalmol ) 2008;8(9):1743–1746.
    1. Ma X, Shimmura S, Miyashita H, Yoshida S, Kubota M, Kawakita T, Tsubota K. Long-term culture and growth kinetics of murine corneal epithelial cells expanded from single corneas. Invest Ophthalmol Vis Sci. 2009;50(6):2716–2721. - PubMed

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