家兔自体虹膜色素上皮移植细胞制备的研究

目的探讨自体虹膜色素上皮（iris pigment epithelium，IPE ）细胞移植时虹膜取材的较佳方法。方法在20只黑兔眼上按人眼周边虹膜切除术的步骤，分别在左眼（ A法，在12点处切取底边为4～5 mm的三角形虹膜组织1块）、右眼（B法，在11点和1点 两处切取底边为2～2.5 mm的三角形虹膜组织块2块）取材，采用酶显微解剖酶分离法对IPE 细胞进行精细的分离和培养，并用光学显微镜、电子显微镜和免疫组织化学等方法对培养的细胞进行观察和鉴定。结果A法取材的IPE细胞培养成功率为65%，B法取材的成功率为95%，经 3～4代的传代即达到移植所需的细胞数。显微镜及免疫组织化学方法观察结果显示，培养的IPE细胞基本保持了原代细胞的特性，细胞活力为85%～93%。结论B法取材的 IPE细胞培养成功率高于A法，培养的IPE细胞在传代到第3代时即可进行移植。（中华眼底病杂志，2003，19：201-268）

Methodological study of preparing iris pigment epithelial cells for autologous transplantation in rabbits

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 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.