新生大鼠视网膜神经元的改良培养及鉴定

目的探讨新生大鼠视网膜神经元获取和体外培养的方法，为视网膜神经细胞的基础研究提供合适的模型。方法采用酶消化法获取生后7—8天的SD（Sprague—Dawley）乳鼠的视网膜神经细胞，使用DMEM／F12培养液在体外培养，倒置显微镜观察细胞形态，免疫组织化学鉴定细胞类型。结果0．25％胰蛋白酶和0．02％EDTA联合消化能使视网膜神经细胞分散，获得单细胞悬液；多聚赖氨酸包被培养皿有助于神经细胞贴壁生长；体外培养第7天的视网膜神经细胞中，视网膜神经元占80％左右，其中50％以上为光感受器细胞。结论酶消化法原代培养视网膜混合神经细胞简单、可行，为视网膜神经细胞的基础研究提供良好的体外模型。

Improved culture and identification of retinal neurons of neonate rat

Objective To investigate the methods of isolation and cultrure neonate rat retinal neurons, provide an appropriate model for the fundamental research of retinal neural cells. Methods Retianl neural cells of postnatal 7 - 8 days rat were obtained by enzyme digestion, cultured in media of DMEM/F12, investigated by inverted microscope and immunohistocbem- istry stain. Results Retinal neural cells could be dispersed as unieall suspension by 0. 25% trypsin and O. 02% ethylene diamion tetra acetic acid （EDTA） digestion. Retinal neurons cultured well on polylysine-treated plated. Among the postcultrured 5 days retinal cells, about 80% is retinal neurons. Among the retinal neurons more than 50% is phototreceptors. Conclusion Primary culture of neonateal rat retinal neurons by enzyme digestion is a simple and feasible method, which provide an appropriated model for the fundamental research of retinal neural cells.