大鼠脑微血管内皮细胞的原代培养与鉴定

目的探讨大鼠脑微血管内皮细胞培养方法。方法 5～7 d龄SD大鼠脑皮质用胶原酶消化得到脑微血管段后,接种于培养瓶进行原代培养。培养的细胞采用倒置显微镜进行形态学观察,免疫荧光法鉴定Ⅷ因子相关抗原,MTT法测定细胞活力。结果倒置显微镜下细胞呈单层＂铺路石样＂排列的典型特征;Ⅷ因子相关抗原免疫荧光检测得阳性细胞率达95%;MTT法测定结果显示第120 h细胞活力最强。结论该培养方法能成功地分离并培养出纯度较高的大鼠脑微血管内皮细胞,对更深入地研究脑微血管内皮细胞的生物学特性及功能具有重要意义。

Primary Culture and Identification of Rat Brain Microvascular Endothelial Cells

Objective To investigate the primary culture system of rat brain microvascular endothelial cells（RBMECs）.Methods Brain microvessel fragments isolated from 2-7-day-old SD rat brain cortex by collagenase digestion were placed on culture flasks.The shapes of cultured cells were observed by the inverted microscope.The Ⅷ factor related antigen was used to identify these cells by the immunofluorescence examination.The cell viability was tested by MTT assay.Results Cultured cells displayed a monolayer growth with a typical ＂cobblestones＂ shape.The result of immunofluorescence test showed 95% of RBMECs were positive.The MTT result indicated the cells growing 120 hours after 3 passages had the strongest cell viability.Conclusion This culture system can highly purify and culture RBMECs.It＇s important to deeply study the biological characteristics and functions of brain of brain microvascular endothelial cells.