Effects of Damaged Human Corneal Epithelial Cells on Differentiation of Human Mesenchymal Stem Cell

ABSTRACT

PURPOSE: To identify the effects of microenvironmental changes caused by human corneal epithelial damages to characteristics or differentiation of human mesenchymal stem cells (hMSCs). METHODS: Artificial corneal damage was induced onto a cultured monolayer of human corneal epithelial cells. hMSCs were then co-cultured with damaged human corneal epithelial cells (dIHCE). Morphological changes in the co-cultured hMSCs were observed. To elucidate the differentiation of hMSCs into corneal keratocytes or epithelial cells, the expressions of alpha-smooth muscle actin, keratin-3/-12, and E-cadherin were confirmed by immunofluorescence. RESULTS: hMSCs co-cultured with dIHCE showed enhanced adherence in the neighborhood of dIHCE and morphological change into dendritic shapes at 6 days post-seeding. Although the expression of alpha-smooth muscle actin, known as hMSCs marker, significantly decreased at the dIHCE-contacted site of hMSCs; there were no expressional changes on keratin-3/-12 and E-cadherin, the markers of corneal epithelial cells. Interestingly, positive expression of corneal epithelial marker keratin-3/-12 was observed in dIHCE co-cultured hMSCs. hMSCs co-cultured with normal human corneal epithelial cells (nIHCE) were unable to attach, and showed no change in the expression of alpha-smooth muscle actin. CONCLUSIONS: It is proposed that dIHCE causes a morphological change in hMSCs, and decreased expression of alpha-smooth muscle actin. These results suggest that dIHCE can affect a change in the characteristics and differentiation of hMSCs.