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Background: This study was undertaken to establish the characterization of cultured oral mucosal epithelium and introducing them as an alternative source for reconstruction of ocular surface disease., Methods: Human oral epithelial cells were cultured on simple media (DMEM/HF12) as control and co-cultured on mitomycin C-treated 3T3 feeder layer, on the amniotic membrane (AM) without nitrocellulose and the mitotically inactivated 3T3 fibroblast, and on the sandwich layer of AM fastened on the nitrocellulose as insert and 3T3 fibroblast. After 3 weeks, the characteristics of the cells were assessed morphologically and also ultrastructurally using scanning electron microscopy and transmission electron microscopy and immuno-cytochemically., Results: The epithelial cells were cultured on AM spread on nitrocellulose insert and 3T3 feeder layer showed better growth than other groups and all groups of study were shown similar characteristics. The cultured oral epithelial shared the characteristics with corneal epithelium., Conclusion: Thus the oral epithelial could be an alternative source for transplantation.