血脑屏障(BBB)调节分子运输,防止病原体,并阻止药物有效输送到大脑。迄今为止的模型未能重现大脑屏障的人体解剖复杂性,导致临床试验中的误导性结果。为了克服这些限制,通过血管发生建立了一种新型的3维BBB微血管网络模型,以准确复制体内神经血管组织.这种微流体系统包括人类诱导多能干细胞来源的内皮细胞,脑周细胞,和星形胶质细胞作为自组装的血管网络在纤维蛋白凝胶。膜转运蛋白的基因表达,紧密连接和细胞外基质蛋白,与几何结构和定量免疫细胞化学的计算分析一致,表明BBB成熟和微环境重塑。共聚焦显微镜验证了微血管-周细胞/星形胶质细胞的动态接触相互作用。BBB模型表现出可灌注和选择性的微脉管系统,渗透性低于传统的体外模型,类似于大鼠大脑的体内测量。这种强大且生理相关的BBB微血管模型为药物发现提供了创新且有价值的平台,可预测临床前应用中的神经治疗转运功效,并概述神经退行性疾病中患者特异性和病理性神经血管功能。
The blood-brain barrier (BBB) regulates molecular trafficking, protects against pathogens, and prevents efficient drug delivery to the brain. Models to date failed to reproduce the human anatomical complexity of brain barriers, contributing to misleading results in clinical trials. To overcome these limitations, a novel 3-dimensional BBB microvascular network model was developed via vasculogenesis to accurately replicate the in vivo neurovascular organization. This microfluidic system includes human induced pluripotent stem cell-derived endothelial cells, brain pericytes, and astrocytes as self-assembled vascular networks in fibrin gel. Gene expression of membrane transporters, tight junction and extracellular matrix proteins, was consistent with computational analysis of geometrical structures and quantitative immunocytochemistry, indicating BBB maturation and microenvironment remodelling. Confocal microscopy validated microvessel-pericyte/astrocyte dynamic contact-interactions. The BBB model exhibited perfusable and selective microvasculature, with permeability lower than conventional in vitro models, and similar to in vivo measurements in rat brain. This robust and physiologically relevant BBB microvascular model offers an innovative and valuable platform for drug discovery to predict neuro-therapeutic transport efficacy in pre-clinical applications as well as recapitulate patient-specific and pathological neurovascular functions in neurodegenerative disease.