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M.-K. Kim, H. Choi, J. Ko, H. Lee, W. Wee, J. Lee; Effect of Human Mesenchymal Stem Cells on Restoration of Homeostasis in Damaged Human Limbal Epithelial Stem Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3745.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate effect of human mesenchymal stem cells (hMSCs) on restoration of homeostasis in chemically-damaged human limbal epithelial stem cells (dhLESCs) through paracrine action.
Primary cultured hLESCs(1.0 X 104 cells/cm2) were damaged with application of 200 µl of 15% ethanol for 30 seconds. Less than 4 passages of Bone-marrow derived human mesenchymal stem cell lines (Lonza, USA) were cultured. To evaluate the paracrine effect of hMSCs on dhLESCs, we cultured dhLESCs for 48 hours in the following conditions; (1) dhLESCs only, (2) dhLESCs with supernatants collected from the hMSC culture (hMSC-conditioned media; hMSC-CM), (3) dhLESCs with supernatants collected from the hMSC cocultured with dhLESC (dhLESC/hMSC-CM). The survival of dhLESCs was estimated with a MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. The homeostasis of dhLESCs were assessed by immunocytochemistry using anti- ABCG2, anti-cytokeratin 3/cytokeratin 12 (CK3/12), and anti-p63 antibody.
The survival of dhLESCs with hMSC-CM and with dhLESC/hMSC-CM was significantly higher than that of dhLESCs only (ANOVA test, p<0.001). dhLESCs with hMSC-CM and with dhLESC/hMSC-CM expressed ABCG2 while, dhLESCs did not express any ABCG2. CK3/12 was diffusely detected in dhLESCs in all conditions.
Through paracrine effect, human MSCs is likely to provide prolonged survival and restore homeostasis of hLESCs even when chemically-damaged.
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