中文摘要：

探讨简易可行的脑微血管内皮细胞（brain microvascular endothelial cells，BMECs）培养方法，为研究BMECs细胞在脑血管疾病中的重要作用提供技术支持。分离出生后1～7天内的SD乳鼠大脑皮质区，植块法培养BMECs细胞。用倒置显微镜观察BMECs细胞的形态以及从皮质块迁出的过程；MTT比色法检测BMECs细胞的生长曲线；采用免疫组化染色检测VⅢ因子相关抗原和CD34抗原，以鉴定内皮细胞。结果发现，大脑皮质块植块法培养的大鼠BMECs细胞呈单层贴壁生长，细胞形态以长梭形、多角形三角形、四边形为主，呈典型的“铺路石”样征象，经鉴定为内皮细胞，第三代纯度达95％以上。提示该方法具有经济、简便、要求条件不高，易于纯化的优点，可作为大鼠BMFCs细胞体外培养的良好模型。

英文摘要：

To explore rat brain microvascular endothelial cells （RBMECs） culture model, we used cortex explant to culture RBMECs and observed the morphology of endothelial cells. After relatively pure cortex explant were obtained from 24 h to 1 weeks old SD rats by careful dissection, cortex explants were putted into the culture flask coated by 1% gelatin. After 2 h cultured in 37 ℃, 5%CO2 incubator, we move a bit of RPMI 1640 culture solution into the culture flask including 90 U/ml heparin sodium, 20% fetal calf serum （FCS）, 100 IU/ml penicillin, 100 μg/ml streptomycin, 4 mmol/L Gln-glutamine and continue to culture. We found that the cultured cells began to migrate from cortex explant after 24 hours, showed the spindle-shaped morphology and reached the monolayer confluence after 10-14 days. Attachment and growth of the cultured cells were depended on the substrata gelatin provided and high-quailty FCS. Immunocytochemistry demonstrated more than 95% of the third generation cultured cells were positive for VIII factor associated antigen and CD34 antigen, which indicated the cultured cells were vascular endothelial cells. The results indicated that relatively pure cortex explant of primary culture of RBMECs was successfully established, which is more simple and cheap than other methods. This method and the model system could be applicable to the studies of physiology, biochemistry and pharmacology of the brain endothelium and could also be used to the studies of pathophysiology of cerebrovascular disease.