Reconstructed corneal epithelium-stroma producted in vitro by tissue engineering

Objective: To reconstruct corneal tissue with epithelium and stroma by tissue engineering.Methods: Epithelial cells and keratocytes were isolated from rabbit corneas and cultured on plastic substrates in vitro They could be subcultured for several passages Using a special transwell, we succeeded to culture two kinds of cells without mixture each other Anti vementin and anti keratin antibodies were used to identify the cells Reconstructed corneal tissues were obtained by culturing epithelial cells on collagen gels containing keratocytes The topmost layer, Consisting of corneal epithelial cells was treated by what was designed as the air liquid interface method The tissue was prepared for histological at 36 h,72 h,7 d,14 d and 20 d.Results: The corneal epithelium and keratocytes cultured in the collagen gel were different from those cultured on plastic morphologically and biochemically, which showed appropriate histological structure Conclusions: The tissue engineered cornea in vitro provides ideal toolsfor further physiological, toxicological and pharmacological studies as well as being an attractive model for gene expression studies