Differentiation of mesenchymal stem cells induced by co-culture method / 中国医学科学院学报

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the potential differentiation of human mesenchymal stem cells(MSCs)into epithelium-like cells by an in vitro co-culture method.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>MSCs can differentiate into epithe1ium-like cells after having been co-cultured with human conjunctival epithelium.</p>