兔视网膜Müller细胞原代培养  

Primary culture of rabbit retinal Mueller cell

作　　者：过贵元[1] 胡淑鸾[2] 

机构地区：[1]福建医科大学教学医院 [2]南京军区福州总医院476临床部眼科

出　　处：《中华眼底病杂志》2011年第2期167-169,共3页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的建立成年兔视网膜Mueler细胞体外原代培养方法。方法运用组织块培养法。分离出成年兔视网膜神经感觉层，剪成1mm×1mm大小组织块，置于含有20％胎牛血清的Dulbecco改良Eagle培养液／F12培养，采用倒置相差显微镜、透射电子显微镜观察及荧光免疫组织化学染色的方法进行培养细胞的鉴定。结果培养的细胞块3d后可见部分细胞突起长出，7d后整个组织块周围放射状长出较多细胞。倒置相差显微镜下，培养细胞呈一端细胞体丰富膨大，另一端有一长突起，细胞核椭圆形，常有2个或2个以上核仁；透射电子显微镜下，细胞胞体大，细胞浆丰富，内含丰富的8～10nm的微丝。荧光免疫组织化学法显示细胞呈神经胶质纤维酸性蛋白和胞内视黄醛结合蛋白阳性，阳性率达95％以上。结论运用组织块培养法可培养出兔视网膜Mueiler细胞。Objective To develop a method for the primary culture of retinal Mueller cells of adult rabbit in vitro. Methods Retina was isolated from adult rabbit, cut into 1 mm × 1 mm pieces, and placed into Dulbecco modified Eagle medium/F12 containing 20% fetal bovine serum to culture. Cultured cells were identified by inverted phase contrast microscope, transmissim electron microscope and immunohistochemistry staining method. Results Visible cell processes grew out from the retinal tissues after three days culture, and more cells grew radically around the retina after seven days culture. The cultured cells were often inflated at one side and had one long process at another side, and the nuclei were elliptical and there were two or more than two nucleoli under inverted phase contrast microscope. The cytoplasm was rich and contained abundant microfilaments in eight to ten nanometers under transmission electron microscope. Immunohistochemistry assay showed that 950/oo of the cells were positive for glial fibrillary acidic protein and cellular retinaldehyde binding protein. Conclusion Rabbit retinal Mueller cells can be cultured by the explant culture method.

关 键 词：细胞培养技术/方法 视网膜/病理生理学 MUELLER细胞 

分 类 号：R774.1[医药卫生—眼科]