大鼠脑微血管内皮细胞的原代培养与鉴定  

Methods of Culturing and Identifying Brain Microvascular Endothelial Cells in Rats

作　　者：吴秀芹[1] 梅晓云[1] 吴颢昕[1] 李洋[1] 李辉[1] 

机构地区：[1]南京中医药大学,南京210046

出　　处：《中国实验方剂学杂志》2011年第3期187-189,共3页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省自然科学基金项目(BK2007243);江苏省"六大人才高峰"资助项目

摘　　要：目的:探索纯度较高、相对经济简便的大鼠脑微血管内皮细胞分离和原代培养的方法。方法:取1～10 d SD大鼠脑皮质经胰蛋白酶消化后,通过不同孔径的筛网过滤,再用胶原酶2次消化获得的微血管内皮细胞进行培养,采用相差显微镜形态学观察及Ⅷ因子相关抗原免疫组化鉴定。结果:1～2 d细胞呈区域性单层贴壁生长,7～9 d呈典型的铺路卵石样征象,Ⅷ因子相关抗原免疫组化检测内皮细胞表达阳性,可见细胞的胞浆及核周呈棕黄色,胞核呈阴性,阳性细胞占绝大部分。结论:采用2次消化、2次过滤技术可以成功分离培养出较理想的大鼠脑微血管内皮细胞。Objective：To explore the methods of culturing highly purified rat brain microvascular endothelial cells（BMEC）. Method： BMEC of SD rats were prepared by filtering through a mesh, and digested by collagenase. Then the cells were examined under microscope and the expression of factor Ⅷ polyclone antibody was determined. Result： The cells became polygon arranging as single layer in 1-2 clays. In 7-10 days, the cells arranged as ovumlike in compact monolayer. Immunocytochemistry of factor Ⅷ polyclone antibody showed positive in cytomembrane and cytoplasm, but negative in cell nucleus. Most of the cells were found to express factor Ⅷ-related antigen. Conclusion： Ideal rat brain microvascular endothelial cells can he cultured by the improved cultural method of double filtering and digestion.

关 键 词：微血管内皮细胞 细胞培养 SD大鼠 

分 类 号：R285.5[医药卫生—中药学]