A microfluidic model of human brain (μHuB) for assessment of blood brain barrier
A microfluidic model of human brain (μHuB) for assessment of blood brain barrier
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DOI：
10.1002/btm2.10126
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发表时间：
2019-05-01
影响因子：
7.4
通讯作者：
Mitragotri, Samir
中科院分区：
文献类型：
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作者：
Brown, Tyler D.;Nowak, Maksymilian;Mitragotri, Samir
微流控细胞模型，通常被称为“芯片上的器官”，通过开发准确和更高通量的模型，继续推动生物工程领域的发展，抓住人体活器官的本质。此类模型可以以传统二维体外模型无法实现的方式模拟关键的体内特征，包括剪切应力和细胞结构。尽管取得了这些进展，但当前的器官芯片模型通常过于复杂，需要高度专业化的设置和设备，并且缺乏轻松确定化合物跨细胞屏障转运动力学的时间和空间差异的能力。为了应对这一挑战，我们报告了在商用微流体平台中使用人脑微血管内皮细胞（hCMEC/D3）和原代人星形胶质细胞开发三维人血脑屏障（BBB）微流体模型（mu HuB）。在mu HuB内，hCMEC/D3单层在24小时内承受生理相关的剪切应力（2.73 dyn/cm(2)）并形成完整的内腔，类似于体内毛细血管。 mu HuB 内的单层表达表型紧密连接标记（Claudin-5 和 ZO-1），这些标记在存在血流动力学样剪切应力后表达增加。当单层细胞静态培养、适应剪切应力并进行非荧光右旋糖酐 (70 kDa) 转运研究时，观察到的细胞损伤可以忽略不计。 mu HuB 的尺寸选择性渗透性为 10 和 70 kDa 葡聚糖，与其他 BBB 模型类似。然而，凭借探测溶质分布的时间和空间演变的能力，mu HuB 能够捕获跨细胞单层的渗透性随时间的真实变化，并允许评估渗透性的全范围，否则使用传统的终点采样技术会丢失这些渗透性。总体而言，mu HuB 平台提供了一个简化、易于使用的模型，可以进一步实时研究人类 BBB 的复杂性，并且可以轻松调整以纳入神经血管单元及其他单元的其他细胞类型。
Microfluidic cellular models, commonly referred to as "organs-on-chips," continue to advance the field of bioengineering via the development of accurate and higher throughput models, captivating the essence of living human organs. This class of models can mimic key in vivo features, including shear stresses and cellular architectures, in ways that cannot be realized by traditional two-dimensional in vitro models. Despite such progress, current organ-on-a-chip models are often overly complex, require highly specialized setups and equipment, and lack the ability to easily ascertain temporal and spatial differences in the transport kinetics of compounds translocating across cellular barriers. To address this challenge, we report the development of a three-dimensional human blood brain barrier (BBB) microfluidic model (mu HuB) using human cerebral microvascular endothelial cells (hCMEC/D3) and primary human astrocytes within a commercially available microfluidic platform. Within mu HuB, hCMEC/D3 monolayers withstood physiologically relevant shear stresses (2.73 dyn/cm(2)) over a period of 24 hr and formed a complete inner lumen, resembling in vivo blood capillaries. Monolayers within mu HuB expressed phenotypical tight junction markers (Claudin-5 and ZO-1), which increased expression after the presence of hemodynamic-like shear stress. Negligible cell injury was observed when the monolayers were cultured statically, conditioned to shear stress, and subjected to nonfluorescent dextran (70 kDa) transport studies. mu HuB experienced size-selective permeability of 10 and 70 kDa dextrans similar to other BBB models. However, with the ability to probe temporal and spatial evolution of solute distribution, mu HuBs possess the ability to capture the true variability in permeability across a cellular monolayer over time and allow for evaluation of the full breadth of permeabilities that would otherwise be lost using traditional end-point sampling techniques. Overall, the mu HuB platform provides a simplified, easy-to-use model to further investigate the complexities of the human BBB in real-time and can be readily adapted to incorporate additional cell types of the neurovascular unit and beyond.