Objective To explore a better method in obtaining iris pigment epithelium(IPE) specimen for autologous transplantation in rabbits. Methods IPE was obtained from 20 black rabbits with method A,i.e.surgical peripheral iridectomy at 12:00 position obtaining a triangle iris tissue with the hemline of 4-5 mm in left eyes,and method B,i.e.surgical peripheral iridectomy at 11:00 and 1:00 positions obtaining two triangle iris tissues with the hemlines of 2-2.5 mm in right eyes . The IP E cells were isolated precisely with enzyme microdissection-enzyme isolation method, cultured in vitro, observed with light and electronic microscope, and ident ified with immunocytochemical staining.ResultsThe success ra te of cells culture were 65% for method A and 95% for method B. After 3-4 generations of culturing,the amount of IPE cells was enough for transplantation, and most of the functions of primary clutured IPE cells were kept still. Viability of IPE cells was 85%-93%. Conclusion The success rate of cells culture for method B is higher than that for method A. The third generation of cultured cells is available for autologous transplantation.(Chin J Ocul Fundus Dis,2003,19:201-268)