The phenotype of in vitro reconstituted normal human oral epithelium is essentially determined by culture medium

OBJECTIVE: To evaluate the role of various culture media and serum supplement on growth of oral cells in monolayer, and on morphogenesis of in vitro reconstituted normal human oral epithelium. METHODS: Primary keratinocytes and fibroblasts were isolated from normal human buccal mucosa. The monolayers were assessed by growth curve analysis and morphology. The organotypic cultures were evaluated by morphometry, immunohistochemistry, and TUNEL. RESULTS: FAD medium (a 3:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium) was able to support fibroblast growth in defined conditions, and to diminish the negative effect of physiological Ca concentration on keratinocytes in monolayers. Medium type had a profound influence on morphogenesis of in vitro reconstituted human oral epithelium. FAD medium was superior to other types of medium tested in supporting both epithelial growth and differentiation. Defined conditions supported epithelial morphogenesis equally well as serum-containing medium. CONCLUSIONS: This study points to an essential role of medium composition for optimized growth and differentiation of primary organotypic cultures.