Evaluation of Barrier Integrity Using a Two-Layered Microfluidic Device Mimicking the Blood-Brain Barrier.
Evaluation of Barrier Integrity Using a Two-Layered Microfluidic Device Mimicking the Blood-Brain Barrier.
复制标题
使用模拟血脑屏障的两层微流体装置评估屏障完整性。
DOI：
10.1007/978-1-0716-3429-5_7
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Cucullo,Luca
中科院分区：
文献类型：
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作者：
Kadry,Hossam;Cucullo,Luca
血脑屏障(BBB)通过控制维持神经元代谢活动和功能所必需的生物物质的流入和流出，在维持脑微环境的动态平衡方面起着至关重要的作用。这种屏障是由不同的细胞在脑微毛细血管的血脑交界处建立的。这些包括微血管内皮细胞、星形胶质细胞和周细胞，以及其他成分，如小胶质细胞、基底膜和神经细胞一起形成的通常被称为神经血管单位；不同的体内和体外平台可用于研究血脑屏障，每个系统提供特定的优点和缺点。最近，芯片上器官平台将微工程技术的优雅与生物系统的复杂性结合在一起，为各种疾病和器官创造了近乎理想的实验模型。这些具有微米大小通道的微流控设备允许细胞在更具生物相关性的环境中生长，使细胞之间能够以组织式的方式连续沐浴在生物液中进行细胞间通信。它们还通过重述机械力和血管灌流来密切代表组织和器官的功能。在这里，我们描述了利用微流控芯片器官技术建立的人源化血脑屏障模型的使用，其中人脑微血管内皮细胞(BMEC)与原代人周细胞和星形胶质细胞共培养。我们详细描述了使用微流控芯片和不同大小的标记葡聚糖作为通透性标志物来评估血脑屏障完整性的方法。此外，我们还提供了一个关于如何在显微镜下研究hBMEC之间紧密连接蛋白表达的详细方案。
The blood-brain barrier (BBB) plays an essential role in maintaining the homeostasis of the brain microenvironment by controlling the influx and efflux of biological substances that are necessary to sustain the neuronal metabolic activity and functions. This barrier is established at the blood-brain interface of the brain microcapillaries by different cells. These include microvascular endothelial cells, astrocytes, and pericytes besides other components such as microglia, basal membrane, and neuronal cells forming together what is commonly referred to as the neurovascular unit; different in vivo and in vitro platforms are available to study the BBB where each system provides specific benefits and drawbacks. Recently, organ-on-a-chip platforms combine the elegance of microengineering technology with the complexity of biological systems to create near-ideal experimental models for various diseases and organs. These microfluidic devices with micron-sized channels allow the cells to be grown in a more biologically relevant environment, enabling cell to cell communications with continuous bathing in biological fluids in a tissue-like fashion. They also closely represent tissue and organ functionality by recapitulating mechanical forces as well as vascular perfusion. Here, we describe the use of humanized BBB model created with microfluidic organ-on-a-chip technology where human brain microvascular endothelial cells (BMECs) are cocultured with primary human pericytes and astrocytes. We thoroughly described the method to assess BBB integrity using a microfluidic chip and various sizes of labeled dextran as permeability markers. In addition, we provide a detailed protocol on how to microscopically investigate the tight junction proteins expression between hBMECs.