一种体外培养兔眼色素上皮细胞的新方法  

A new method of culturing rabbit retinal pigment epithelial cells in vitro

作　　者：代丽丽[1] 丛丽丹[2] 孙大卫[1] 焦万秋 张中宇[1] 彭绍民[1] 

机构地区：[1]哈尔滨医科大学第二临床医学院眼科,黑龙江哈尔滨150081 [2]哈尔滨医科大学第一临床医学院眼科,黑龙江哈尔滨150001 [3]哈尔滨市眼科医院眼科,黑龙江哈尔滨150001

出　　处：《哈尔滨医科大学学报》2009年第4期336-338,共3页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目(30271395);国家自然科学基金资助项目(30772381);黑龙江省自然科学基金资助项目(D0241);哈尔滨医科大学第二临床医学院青年基金资助项目(QN200603)

摘　　要：目的建立一种体外培养的兔眼视网膜色素上皮(retinal pigment epithelium,RPE)细胞的新方法。方法采用Dispase对兔眼RPE细胞进行原代培养,免疫组化通过MNF116和s-100鉴定细胞纯度。将细胞接种于铺有laminin的Transwell滤膜上,观察到细胞融合后对膜切面行透射电子显微镜观察;并通过细胞ZO-1免疫荧光化学法了解紧密连接的形成情况。结果成功培养了原代兔眼RPE细胞并且无其它细胞污染。透射电镜可见细胞表面有大量微绒毛并形成紧密连接。ZO-1免疫荧光化学结果可见细胞形态基本呈多边形,紧密连接基本形成。结论兔眼RPE细胞能够用Dispase成功分离并培养。Objective To develop a new method of culturing retinal pigment epithelium （RPE） cells in vitro. Methods Primary rabbit RPE cells were isolated with dispase and cultured. Immunochemistry of MNF116 and S-100 was performed to identify the purity of the cells. The section of filter was observed by transmission electron microscope. Immunofluorescence was performed to label the ZO-1 protein of tight junction. Results Primary adult rabbit RPE cells were cultured successfully and were not contaminated by other cells. Transmission electron microscope results demonstrated the cultured cells were excellent. ZO-1 expression in good circumferential bands demonstrated that the RPE formed possible junctional complexes. Conclusion The adult rabbit RPE cells can be isolated with Dispase and cultured successfully.

关 键 词：视网膜色素上皮 细胞培养 紧密连接 

分 类 号：R332[医药卫生—人体生理学]