Methodological study of preparing iris pigment epithelial cells for autologous transplantation in rabbits / 中华眼底病杂志

ABSTRACT

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 1200 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 1100 and 100 positions obtaining two triangle iris tissues with the hemlines of 2-2.5 mm in right eyes . The IPE cells were isolated precisely with enzyme microdissection-enzyme isolation method, cultured in vitro, observed with light and electronic microscope, and identified with immunocytochemical staining. Results The success rate 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.