Objective To explore the feasibility of the transdifferentiation of human mesenchymal stem cell (MSC) into corneal epithelial cells in vitro. Methods Experimental study.The third passage human MSC and the third passage corneal stromal fibroblasts were co-cultured in a Transwell culture system for 1 week. The human MSC were observed and identified by light microscopy, scanning and transmission electron microscopy, and comprehensive indirect immunocytochemistry staining in the induced and control groups. Results Both the third passage human MSC and the third passage corneal stromal fibroblast cells could adhere and grow rapidly.After co-culture with corneal stromal fibroblasts for 1 week, the morphology of some induced human MSC changed into epithelial-like cells. Some induced human MSC showed positive staining for monoclonal antibody AE1, which were typical epithelium cells with a large quantity of microvilli,desmosomes and tonofilaments that were observed under electron microcopy. Conclusion The data suggests that human MSC may have the potential to transdifferentiate into human corneal epithelial-like cells in vitro.