摘要
贴块法培养脑微血管内皮细胞(BMECs),倒置显微镜动态观察细胞生长及形态,Ⅷ因子相关抗原、CD34免疫细胞化学联合鉴定细胞并确定纯度。免疫细胞化学和Western印迹法检测药物转运体有机阴离子转运多肽亚型2(Oatp2)及P-糖蛋白(P-gp)在培养内皮细胞上的表达。结果显示,获得的BMECs呈多角形或铺路石形,单层贴壁生长;培养细胞Ⅷ因子相关抗原免疫细胞化学、CD34免疫荧光染色均为阳性,细胞纯度90%;培养细胞有Oatp2及P-gp表达,且二者均主要表达于BMECs细胞膜。提示贴块法可获得原代培养BMECs,方法简便易行,细胞纯度较高。原代培养的BMECs上有药物转运体Oatp2及P-gp的表达,为血脑屏障上药物转运体的体外研究提供了可能途径。
Rat brain microvascular endothelial cells （BMECs） were cultured with brain segment sticking mass methods. The BMECs were identified by observing under phase-contrast microscope, and by using immunocytochemical methods with anti-rat factor Ⅷantibody and CD34.The expression of organic anion transporting polypeptide2 （Oatp2） and P-glycoprotein （P-gp） were detected by immunocytochemical methods and Western blot analysis. In this study, the BMECs showed regular cobblestone and polygonal morphology and positive for ⅧFAg and CD34. The purity of positive stained cells was 90%. Both Oatp2 and P-gp expressed in the primary BMECs, and preminanfly along the plasma membrane of endothelial cells. It is suggested that the rat BMECs were successfully cultured in vitro by brain segment sticking mass methods. The primary BMECs expressed the Oatp2 and P-gp. This experiment provide a possible pathway for studying transporters of the blood-brain barrier.
出处
《细胞生物学杂志》
CSCD
2007年第4期560-564,共5页
Chinese Journal of Cell Biology
基金
重庆市教委(No.KT060305)
重庆市卫生局(渝卫科教[2006]34号06-2-157)资助项目~~
