目的探讨人胚眼视网膜光感受器细胞的获取和体外培养方法，为进行人类视网膜光感受器细胞移植提供理论依据。方法采用酶分层消化法分离获取人胚眼纯视网膜光感受器细胞，使用视网膜色素上皮细胞的条件培养液，在体外长期培养并进行形态学鉴别；用苏木素-伊红染色显示消化后的视网膜组织结构层次，以确定所获取的视网膜光感受器细胞的纯度；用抗视杆细胞特异性视紫质蛋白抗体（rhodopsin4D2，Rho4D2）免疫细胞化学染色做细胞鉴定。结果酶分层消化法能获取较纯的视网膜光感受器细胞，抗Rho 4D2阳性，并能在体外存活较长时间。结论酶分层消化法所获取的人胚眼纯视网膜光感受器细胞能在体外较长时间存活，并可表达光感受器细胞特异性视紫质蛋白，将为人类视网膜光感受器细胞的移植和各种体外研究提供细胞来源。%Objective To investigate the methods of isolation and culture of fetal human retinal photoreceptors. Methods Fetal human retinal photoreceptors were obtained by enzyme gradation digestion, cultured in media conditioned by retinal pigment epithelial cells (RPE-CM), identified by morphologic observation and immunohistochemically stained with rod specific opsin 4D2 (Rho 4D2) antibody. The digested retinal tissues were tested by hematoxylin-eosin (HE) staining. Results Enzyme gradation digestion could obtain relatively pure retinal photoreceptors. These cells were stained positively for Rho 4D2 antibody, and survived in RPE-CM for a long time. Conclusion Fetal human retinal photoreceptors obtained by enzyme gradation digestion can survive in RPE-CM for a long time and express rod specific opsin. This culture system will provide a source for human retinal photoreceptor transplantation and further studies of photoreceptors in vitro.
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