In vitro study of culture of oral mucosa basal cells

Objectives:One of the keys of the oral tissue engineering is to obtain adequate living oral mucosa basal cells (OMBCs) in a comparatively short time. Currently, the culture period of OMBCs in vitro is too long to meet the needs of oral tissue engineering. The objective is to study the optimal procedure for the culture and amplification of OMBCs. Methods:OMBCs were digested by dispase together with trypsin. Isolated cells were divided into four groups: Group A was cultivated by DMEM with10% FBS; Group B was cultivated by DMEM with10% autogenous serum (AS); Group C was cultivated by MCDB153 serum free medium; Group D was cultured by two-step culture method (primary culture medium was DMEM with10% AS, but subculture medium was changed into MCDB153 serum free medium). The biological characteristics of the cells were observed through reverse phase-contrast microscopy, and the ratio of adhesion, cell growth curve, population-doubling time (PDT) were recorded. Results:Group D could shorten the culture time obviously and enhance the viability of OMBCs. Conclusions:the two-step culture method not only exerted strongpoint of the media supplemented with serum in primary culture, but also reserved the advantage of the serum-free media in subculture. So two-step culture method could supply enough cells for oral mucosa tissue engineering. Supported by Province Health Hall Foundation of Guangdong. WSTJJ2001112242010619530501494