PDF(Pubmed)
Abstract：
Blood-brain barrier (BBB) dysfunction is a key feature in neuroimmunological and neurodegenerative diseases. In this study, we developed a microfluidic human BBB-on-a-chip to model barrier dysfunction and immune cell migration using immortalized TY10 brain endothelial cells, pericytes, and astrocytes. It was found that immortalized TY10 brain endothelial cells developed a microvascular structure under flow. Pericytes were localized on the basal side surrounding the TY10 microvascular structure, showing an in vivo-like structure. Barrier integrity increased under co-culture with pericytes. In addition, both ethylenediaminetetraacetic acid (EDTA) and anti-Claudin-5 (CLDN5) neutralizing antibody caused a decrease in the transendothelial electrical resistance (TEER). EDTA caused the leakage of 20 kDa dextran, suggesting different effects on the BBB based on the mechanism of action, whereas anti-CLDN5 antibody did not cause leakage. In the tri-culture model, human T cells migrated through endothelial vessels towards basal C-X-C motif chemokine ligand 12 (CXCL12). The live-imaging analysis confirmed the extravasation of fluorescence-labelled T cells in a CXCL12-concentration- and time-dependent manner. Our BBB model had an in vivo-like structure and successfully represented barrier dysfunction and transendothelial T cell migration. In addition, our study suggests that the inhibition of CLDN5 attenuates the BBB in humans. This platform has various potential uses in relation to the BBB in both drug discovery research and in elucidating the mechanisms of central nervous system diseases.
摘要：
血脑屏障（BBB）功能障碍是神经免疫和神经退行性疾病的关键特征。在这项研究中,我们开发了一种微流控人类BBB-on-a-chip，使用永生化的TY10脑内皮细胞来模拟屏障功能障碍和免疫细胞迁移，周细胞,和星形胶质细胞。发现永生化的TY10脑内皮细胞在流动下形成微血管结构。周细胞位于TY10微血管结构周围的基底侧，表现出类似体内的结构。与周细胞共培养时，屏障完整性增加。此外,乙二胺四乙酸（EDTA）和抗Claudin-5（CLDN5）中和抗体均导致跨内皮电阻（TEER）降低。EDTA导致20kDa葡聚糖的泄漏,根据作用机制提示对血脑屏障的不同影响，而抗CLDN5抗体不引起渗漏。在三文化模型中，人T细胞通过内皮细胞向基础C-X-C基序趋化因子配体12(CXCL12)迁移。活成像分析证实了荧光标记的T细胞以CXCL12浓度和时间依赖性方式的外渗。我们的BBB模型具有体内样结构，并成功代表了屏障功能障碍和跨内皮T细胞迁移。此外,我们的研究表明，CLDN5的抑制减弱了人类的BBB。该平台在药物发现研究和阐明中枢神经系统疾病的机制中具有与BBB有关的各种潜在用途。
