体外培养的成年兔眼色素上皮细胞屏障功能的研究  

Barrier function of adult rabbit retinal pigment epithelial cells cultured in vitro

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

机构地区：[1]哈尔滨医科大学附属第二医院眼科,黑龙江省哈尔滨市150086

出　　处：《眼科新进展》2008年第3期166-168,共3页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30271395;30772381);黑龙江省自然科学基金资助(编号:D0241);哈尔滨医科大学附属第二医院青年基金资助(编号:QN200603)~~

摘　　要：目的研究体外培养的成年兔眼视网膜色素上皮(retinal pigment epithelial,RPE)细胞的血-视网膜外屏障功能。方法对成年兔眼RPE细胞进行原代培养,免疫组织化学通过MNF116和S-100鉴定细胞纯度。将细胞接种于铺有层粘连蛋白的Transwell滤膜上,分别于培养过程中不同时间点进行跨膜电阻测定,待电阻达到平台期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。结果成功原代培养了兔眼RPE细胞并且无其他细胞污染。接种于Transwell后,RPE细胞TER值3周达到最高值约88.14Ω·cm2,维持1周并开始下降。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。结论成年兔眼RPE细胞通过培养于铺有层粘连蛋白的Transwell滤膜上可以成功建立细胞屏障功能模型。Objective To investigate the blood-retinal barrier function of adult rabbit retinal pigment epithelial（RPE） cells cultured in vitro.Methods Primary rabbit RPE cells were cultured and immunochemistry of MNF116 and S-100 was performed to identify the purity of the cells.The cells were plated on laminin-coated Transwell filter.Transmembrance electric resistance（TER） was detected at different time points.After the TER reached a platform of high level,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.The value of TER reached at highest level 88.14 Ω·cm2 after three weeks and maintained for one week before dropping.Transmission electron microscope results demonstrated that culture cells were excellent.ZO-1 expression in good circumferential bands demonstrated that the RPE formed possible junctional complex.Conclusion The barrier function model of adult rabbit RPE cells can successfully set up with culturing cells on the laminin-coated filter.

关 键 词：视网膜色素上皮 细胞培养 血-视网膜屏障 紧密连接 免 

分 类 号：R774.1[医药卫生—眼科]