Identification and Characterization of Phenotypic Markers of Human Mesenchymal Stem Cell-Derived Corneal Limbal Epithelial Cells

Abstract

Purpose: To investigate the expression of stem cell-associated and corneal differentiation markers in human bone marrow-derived MSCs (hMSCs) and to examine the influence of cell culture conditions on the potential of hMSCs to differentiate into corneal epithelial cells. Methods: We identified and characterized stern cell-associated and corneal differentiation markers in hMSCs using RT PCR and immunocytochemistry, and compared it to the marker expression seen in human corneal limbal epithelial cells (hLECs). We also examined the influence of cell Culture conditions, Such as serial passage, cell plating density, presence of feeder cells, and addition of epidermal (EGF) and fibroblast growth factors (FGF) at varying densities, oil the marker expression of hMSCs. Results: Both hMSCs and hLECs expressed Cnx43. However, hMSCs did not express p63, ABCG2, OCT4, K12, or MUC16, which were expressed in hLECs. MUC1 was highly expressed in hMSCs, while it was not expressed in hLECs. Notably, hMSCs lost the ability to express MUC1, when 20 ng/ml of EGF was added to the culture in the presence of NIH/3T3 feeder cells. However, the Culture duration, cell plating density, and use of feeder cells did not exert a significant effect oil the marker expression in hMSCs. Conclusion: P63, K12, and MUC16 can be used as putative positive markers and MUC1 as a putative negative marker for differentiation of hMSCs to hLECs. Differentiation of hMSCs into corneal epithelial-like phenotype was not induced by varying the culture duration or plating density or by using NIH/3T3 fibroblasts as a feeder.