摘要
目的分离、培养和纯化牛原代视网膜色素上皮细胞(RPE)和虹膜色素上皮细胞(IPE),比较体外培养的RPE与IPE的生物学特性。方法采用酶辅助显微分离牛眼RPE和IPE,进行体外培养,密度梯度离心法纯化并采用免疫组化法进行鉴定。透射电镜观察体外培养RPE、IPE的超微结构,比较两种细胞体外培养下的生长状况。结果纯化后体外培养牛RPE和IPE的角蛋白表达强阳性,细胞阳性率分别达到94%和88.7%。原代培养的牛RPE和IPE外形类似,为多角形,具有相似的生长曲线。电镜显示两种细胞都具有细胞极性,顶端有大量微绒毛,原代细胞内含有大量色素颗粒和内质网,RPE含有较多与其吞噬功能相适应的溶酶体。结论通过酶辅助显微分离色素上皮细胞,结合密度梯度离心的方法可以获得较纯的牛RPE和IPE,该方法同样适用于周切虹膜材料。两种色素上皮细胞具有相似的形态和生物学性状,但仍然存在与功能相适应的超微结构差异。
Objective To investigate the differences in biological characteristics between bovine iris pigment epithelium （IPE） and retinal pigment epithelium （RPE）. Methods IPE and RPE were isolated from bovine eyes using enzyme-assisted micro dissection, and were purified with density gradient centrifugation technique and identified with immunocytochemical staining. The uhrastructure of RPE and IPE was observed by transmission electron microscope, and the cell growth was compared between them. Results There were high positive staining of anticytokeratin in RPE and IPE after purification, 94% and 88.7% , respectively. Both RPE and IPE were polygonal or euboidal with dense pigmentation. The growth curves were similar. It was indicated by transmission electron microscopy that there were mierovilli on the upper surface of the cells and minimal basal lamina production with hemidesmosome-like structure at the base of the cells. There were numerous mature melanosomes in both cells. Fewer lysosomes were observed in IPE compared with RPE. Conclusion IPE resemble RPE in morphology and biological characteristics, while there are differences in function-adaptive ultrastueture between them.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2008年第6期635-639,共5页
Journal of Shanghai Jiao tong University:Medical Science
基金
上海市科委青年科技启明星计划(02QB14037)
上海交通大学医学院优秀中青年基金(040402)~~
