药物实验血脑屏障体外模型的建立及评价

Establishment and evaluation of an in vitro model of brain-blood barrier for pharmaceutical experimental reserch

目的:建立模拟在体血脑屏障(blood-brain barrier,BBB)模型并评价其形态和功能,为中枢神经系统药物的跨血脑屏障特性研究提供体外实验的模型依据。方法:将BALB/c小鼠脑微血管内皮细胞(brain microvascular endothelial cells,BMVEC)和大鼠C6胶质瘤细胞分别接种于经明胶包被的Transwell多聚碳酯膜内室和外室,进行非接触双室共培养,建立细胞培养水平的BBB模型,分别考察模型形成前后BBB光镜、扫描电镜和细胞的跨内皮电阻值(transendothelial electrical resistance,TEERs)。结果:BMVEC培养至汇合后,扫描电镜显示细胞呈单层平铺生长并产生紧密连接,TEERs值较未形成屏障时明显升高(P<0.05)。结论:构建的BBB体外模型在形态学、跨内皮电阻和通透性方面具备了在体BBB的基本特性,适用于中枢神经系统药物跨BBB特性的研究。

OBJECTIVE To establish an in vitro model of brain-blood barrier （BBB） using cultured mouse brain microvascular endothelial cells （BMVEC） and glioma cell line C6 for pharmaceutical research on the trans-BBB drug delivery property. MESULTS Mouse BMVEC were seeded on the surface of polyester-multipore-membrane with gelatin-coated cell culture insert （Transwell） ,C6 cells were seeded on the bottom of cell culture plate for co-cultured to confluence. To evaluate the morphology and characters of BMVEC in themodel, light and scanning electron microscopy were used, and transendotheliar resistance （TEERs） over BMVEC of the barrier was determined with Millicell-ERS system. RESULTS The in vitro BBB model has similar properties as in vivo BBB such as tight junctions between endothelial cells, produce high TEERs values. CONCLUSION The in vitro BBB model at cell level in our study is suitable for central nervous system （CNS） drug trans BBB research.