[Differentiation of mesenchymal stem cells induced by co-culture method]

To investigate the potential differentiation of human mesenchymal stem cells(MSCs)into epithelium-like cells by an in vitro co-culture method.

The human conjunctival epithelium was obtained by digestion with dispase2, and the MSCs were isolated by density gradient centrifugalization. All cells were identified according to their morphologies and cell-surface antigen profiles by immunocytochemical analysis. The MSCs underwent co-culture with conjunctival epithelium in the manner without cell-to-cell contract. The morphological characterizes of cells were observed under contrast microscope, and the cytokeratin-4 expressions of the differentiated cells were identified by immunocytochemistry staining, reverse transcriptase polymerase chain reaction(RT-PCR), and Western blotting.

Immunocytochemistry showed that positive expression of CD29 and negative expression of CD34 in the in vitro cultured MSCs. Cytokeratins4(CK4)was positively expressed in the human conjunctival epithelium. After co-cultured with conjunctival epithelial for 10 days, CK4 was detected in differentiated cells by immunocytochemistry, RT-PCR, and Western blotting.

MSCs can differentiate into epithe1ium-like cells after having been co-cultured with human conjunctival epithelium.