论文部分内容阅读
采用胶原酶消化、差速离心、尼龙网滤过技术分离和获取鼠脑微血管内皮细胞,接种后4h换液使获得的内皮细胞纯化,体外进行长期培养。细胞在体外生长176天,传至30代,细胞初期成活率为92％,纯度近90％。经形态学、超微结构和免疫组化鉴定,培养细胞为血管内皮细胞。培养至第30代的细胞仍能合成和分泌PGI2、ACE等,ⅧF:Ag阳性表达,染色体为二倍体(2n=42),基本保持着细胞的主要特征。该分离和培养方法的建立,将为研究与脑血管相关疾病提供有用工具。
The collagenase digestion, differential centrifugation and nylon mesh filtration technique were used to isolate and obtain rat brain microvascular endothelial cells. The endothelial cells were purified by liquid exchange 4h after inoculation, and cultured in vitro for long term. The cells were grown in vitro for 176 days and passed on to passage 30 for an initial cell viability of 92% and a purity of nearly 90%. The morphological, ultrastructure and immunohistochemical identification of cultured cells for vascular endothelial cells. PGI2, ACE, etc. were still able to be synthesized and secreted by the 30th generation. The positive expression of Ⅷ F: Ag and the diploid chromosome (2n = 42) kept the main characteristics of the cells. The establishment of this separation and cultivation method will provide a useful tool for studying cerebrovascular related diseases.