RECONSTITUTED HUMAN ORAL AND ESOPHAGEAL MUCOSA IN CULTURE

We have successfully established monolayer and organotypic culture tec hniques for growing human oral and esophageal epithelial cells, Cells in monolayer culture were grown in serum-free medium, modified from te chniques previously reported by our group. The organotypic cultures we re grown in a defined medium supplemented with 10% fetal calf serum. O ral and esophageal cells were maintained in keratinocyte basal medium with pituitary extract and otter supplements, and 0.05 mM calcium for 7-9 and 9-11 passages, respective?v. Both cell types had similar morph ology by phase contrast microscopy. When confluent, the cells were pre dominantly small: basaloid, and uniform and interspersed with larger, differentiated felts, By immunohistochemistry, both cell types in mono layer were positive to AE1, AE3, and 34BE12 antibodies to keratins of stratified epithelia. Oral epithelial cells in monolayer also were pos itive to 35BH11, representative of simple epithelial keratins: while e sophageal cells were not. The esophageal cells were focally positive t o K13, while the oral cells were negative, Both were negative for K19. When comparing monolayer to organotypic cultures and to in vivo speci mens, there was a significant difference in the expression of keratins . Using organotypic cultures, AE1, AE3, and 34BE12 were strongly posit ive in both oral and esophageal cells, similar to in vivo tissues. In contrast to monolayers, both were also focally positive for K19. Esoph ageal cells were strongly positive for K13, while the oral cells were mildly but uniformly positive. Both were negative for keratins of simp le epithelia. These two cell culture techniques offer unique opportuni ties to study the pathobiology including carcinogenesis, of stable cel l systems from the oral and esophageal epithelia.