兔眼虹膜色素上皮细胞自体视网膜下移植的研究

目的探讨兔眼虹膜色素上皮(IPE)细胞的自体移植技术并进行术后移植区的细胞形态学观察。方法通过虹膜全切法获取10只新西兰白兔的虹膜,采用酶-机械分离-酶消化的方法分离IPE细胞,用5溴脱氧尿嘧啶(5-Brdu)标记原代或一代的细胞。将标记过的IPE细胞悬液通过内路法分别植入对侧眼的视网膜下腔。术后2、4、6周摘除眼球对移植区进行光镜、电镜检查。结果经免疫学鉴定纯化培养了兔眼IPE细胞。术后光镜显示移植的IPE细胞成单层贴附在原始的视网膜色素上皮(RPE)细胞上。电镜结果证实移植的IPE细胞尖端微绒毛与光感受器外节连接,邻近的光感受器细胞显示正常结构。结论采用内路法可成功地将细胞悬液植入家兔视网膜下腔。6周内移植的自体IPE细胞在视网膜下腔存活,与原先的RPE细胞生长状态类似,邻近的光感受器细胞无形态改变。

Experimental study on transplantation of autologous iris pigment epithelial cell into rabbit subretina space

Objective Iris pigment epithelial(IPE)cells can hopefully replace the autologous transplantation of RPE cells,and can also be used in the treatment of age-related macular degeneration(AMD)and other diseases.Present study aimed to investigate the autologous transplantation of IPE cells of rabbit eyes and observe morphologically transplanted cells in subretina.MethodsTen irises from New Zealand rabbits were obtained during iridectomy surgery,and IPE cells were isolated by the enzyme-mechanic dissection-enzyme digestion method.Primary cultured IPE cells and the first generation of cells were labeled with 5-Brdu.Suspension of autologous IPE cells was injected into the subretinal space of the fellow eyes using the internal approach method.Experimental eyeballs were extirpated 2,4 and 6 weeks later respectively for the examination of light and electron microscope.ResultsIPE cells of rabbit eyes were isolated and incubated in DMEM containing 10?S and were identified and purified using immunological method.After the surgery,transplanted IPE cells formed one contiguous layer on the top of native RPE under the light microscope and cellular apical microvilli connected to the outer segment of the photoreceptor under the electron microscope.No infiltration of lymphocytes or macrophages was found.The adjacent rod outer segments remained regular throughout the experimental period.ConclusionInternal approaches could successfully transplant the cells suspension liquid into the subretinal space of rabbits.The autologous IPE cells could survive and grow similar to the original RPE cells in the subretinal space until 6 weeks after transplantation without influence on abject photoreceptors.