基于微流控芯片的体外血脑屏障模型构建

Construction of Blood-brain Barrier in vitro Model Based on Microfluidic Chip

目的:利用微流控芯片技术构建易调控、接近在体微环境的体外血脑屏障模型。方法:微流控芯片体外模型采用上下双培养池结构,由多聚碳酸酯膜分隔,两套流路系统控制流体。细胞采用原代分离纯化的大鼠脑血管内皮细胞和星形胶质细胞,免疫荧光技术进行鉴定,分别按次序注入微流控芯片上下培养池,按1μl/min的流速进行灌注培养,构建体外血脑屏障模型,并对此模型进行鉴定和评价。结果:原代分离纯化得到两种细胞,免疫荧光法鉴定细胞纯度达95%以上。共培养3天紧密连接开始形成,5天达到峰值,超微结构观察显示内皮细胞之间形成紧密连接,且荧光素钠渗透实验和TEER值测量表明屏障形成良好。结论:成功构建微流控芯片体外血脑屏障模型,可成为一个新的平台应用于药物筛选、神经系统基础等多项研究中。

Objective： To create a blood-brain barrier in vitro model by microfluidic chip technology, which were easy to control and close to microenvironment. Methods： The microfluidic chips were composed of dual pool structure, separated by the polycarbonate membrane, vascular endothelial cells and astrocytes were and there were two sets of flow control system, mouse brain isolated and purified, the cells were identified by immunofluorescence. Then cells were added to the two pools according to the order, and infused at 1 Ixl/min flow rate. The blood-brain barrier （BBB） model was constructed, and then identified and evaluated. Results： Two kinds of cells were obtained by isolation and purification, and the cells were identified by immunofluorescence and the purity was above 95%. After 3 days of co-culture, tight junctions began to develop, and the peak is on the fifth day, the ultrastructural observation showed that the mouse brain vascular endothelial cells formed tight junctions, the penetration experiments of fluorescein sodium and the measurement of TEER showed that barrier formation was good. Conclusion： The blood-brain barrier model was constructed based on the microfluidic chip, It may become a new platform for drug screening, neural system basis and other studies.