牛视网膜微血管内皮细胞体外分离与培养的实验研究  

Isolation and Culture of Bovine Retinal Capillary Endothelial Cells in Vitro

作　　者：张朝云[1] 雷闽湘[1] 

机构地区：[1]中南大学湘雅医院内分泌科,湖南长沙410008

出　　处：《现代生物医学进展》2007年第5期686-688,共3页Progress in Modern Biomedicine

摘　　要：目的:探讨牛视网膜微血管内皮细胞(bovine retinal capillary endothelial cells,BREcs)体外分离、培养方法,为研究视网膜血管性疾病提供一定的实验基础。方法:无菌条件下取出视网膜并剪碎,经筛网过滤、胶原酶消化获取视网膜微血管内皮细胞,接种于明胶包被的培养瓶中,原代培养时用不同的培养基筛选细胞,并在传代时利用差速黏附法以获得较纯BRECs,通过形态学观察和免疫组化方法鉴定BRECs。结果:用此法原代培养BRECs纯度达98%,混有的血细胞及神经组织细胞碎片在换液和传代过程中逐渐被去除,成纤维细胞和周细胞的污染可分别用不同的培养基和差速黏附法纯化去除。结论:该方法简单有效,获得的BRECs纯度高,生长状态良好,为研究眼部血管性疾病提供良好平台。Objective： To explore an improved method for the isolation and culture of bovine retinal capillary endothelial cells （BRECs） in vitro. Methods ：Retinas were detached from the optic nerve and cut into small segments under aseptic condition. After passing through the double filtration with 100urn nylon mesh and collagenase digestion, the suspension was cultured in a gelatin coated dish with DMEM. The primary cultured cells were selected by different medium. Then, differential conglutination was utilized to get pure BRECs at the time of passage. They were then identified by morphological observation and immunocytochemistry studies of factor Ⅷ relative antigen. Results：The blood cells, nerve cells, fibroblasts and pericytes mixed in the primary cultured BRECs were removed gradually in the course of replacing medium and passing. The purity of BRECs reaches up to 98%. Conclusion：The pure BRECs can be attained rapidly and easily by the described methods. These cultures would provide a useful tool for studying diseases related with retinal vessels in vitro.

关 键 词：牛 视网膜微血管 内皮细胞 培养 

分 类 号：Q253[生物学—细胞生物学]