Abstract: Objective: To provide a sound basis for the experimental research by cultivation of endothelial cells from rabbit corneas.Methods: The Descemet’s membrane with its endothelial covering was separated from the underlying stroma by microdissection.Different methods were used to isolate and cultivate corneal endothelial cells of rabbits in vitro and their characteristics were examined.Results: Cultured corneal endothelial cells of rabbits were hexagonal or polygonal and formed a compact monolayer.The cultured cells appeared to maintain the morphologic characteristics of endothelial cells in vivo.The Descemet’s membrane-endothelium from rabbits was cultured for one day at first,and then incubated with trypsin and EDTA in D-Hanks solution.The shortest culture period was about a week with this method.Conclusions: The Descemet’s membrane with its endothelial covering can supply for cultivating the epithelial cells of corneas.