Objective To analyze influential factors of cultural method of human oral mucosal epithelial cells （hOMEC）and to establish a reliable cell culture system for hOMEC. Methods Benzalkonium bromide and gentamicin sulfate were used to prevent microbial contamination. Separations of epithelium from underlying connective tissues with Dispase at different concentration were compared. Enzyme digestion was used to isolate cells and ker - atinocyte- serum free medium（K- SFM）was employed for primary culture and subculture of hOMEC. Results Microbial contamination was under control. Separation of epithelium from underlying connective tissues with 0.40% Dispase was more complete than that of 0.25% Dispase. The cells grew fast and well in vitro. Conclusion The high successful culture of hOMEC and simplified procedures could be obtained with improvement of methods.
