简便有效分离培养大鼠主动脉内皮细胞的一种新方法

A new method for isolation and culture of the endothelial cells of the rat aorta

目的:探索分离、培养鼠主动脉内皮细胞的有效方法。方法:将鼠主动脉内膜翻向外,结扎两端,置于50mL培养瓶中培养、传代,Ⅷ因子相关抗原免疫细胞化学染色。结果:培养6～8d时有少量细胞自主动脉迁移出并贴壁生长;12～14d时贴壁细胞覆盖大部分培养瓶面,约80%的细胞汇合成单层;传代细胞生长旺盛;细胞汇合后呈现典型的"鹅卵石"样内皮细胞特征,内皮细胞标志物Ⅷ因子相关抗原表达阳性;未见杂细胞。结论:此方法无损伤、不需要酶消化,能比较容易的获得高纯度大鼠原代主动脉内皮细胞,适用于细小血管内皮细胞的分离与培养。

Objeetive：To investigate a modified method of isolating and culturing the endothelial cells from the rat aorta for endothelial bioresearch,Methods：Turned inside out, the aorta of the rat,of which the intima tunica was exposed, was ligated at both ends and cultured in a 50 mL culture bottle. Results：A small amount of cells migrated out from the aorta 6 to 8 days after cultured; the migrating cells spread over most of the culture surface of the bottle 12 to 14 days after cultured, 80% of them in a confluent monolayer. The confluent cells were identified as endothelial cells with the typical cobblestone appearance and the positive reaction for factor VII relative antigen, no other types of cells was observed.Cenclassion：This is a woundless, nonenzymatic method of obtaining highly purified primary endothelial cells of the rat aorta, especially practical for isolation and culture of small vascular endothelial cells.