Abstract:

Objective To investigate the feasibility of inducing human dental pulp stem cells (DPSCs) to differentiate into corneal epithelial-like cells by conditioned medium (CM). Methods DPSCs were isolated and identified by flow cytometry. The effect of basal medium (BM) and different CM on the proliferation activity of DPSCs was detected by cell counting kit-8(CCK-8) assay. DPSCs were induced by 30%CM, 60%CM, 90%CM. The cells cultured in BM were negative control group. Corneal epithelial cells markers cytokeratin 3 (CK3) and cytokeratin 12 (CK12) were detected by immunofluorescence assay. Results There was no significant difference in the proliferation activity of DPSCs between BM group and different CM group(P> 0.05). Cells in the 30%, 60%, 90%CM group did not express CK3 after 3 days induction, cells in the 60%and the 90%CM group began to express CK12; CK3 and CK12 were expressed in the 30%, 60%, 90%CM group after 7 days; At the 11th and 14th day, cells continued to express CK3 and CK12 in the 30%, 60%, and 90% CM groups. No expression of CK3 and CK12 was observed in the BM group. Conclusion DPSCs are capable of differentiating into corneal epithelial-like cells under the induction of CM.

Key words: Dental pulp stem cell, Conditioned medium, Flow cytometry, Immunofluorescence, Humen