Abstract：
The blood-brain barrier (BBB), mainly composed of brain microvascular endothelial cells, astrocyte end-feet, and pericytes, serves as a physical and biochemical barrier that selectively limits the passage of circulating molecules into the brain parenchyma. The disruption of its integrity and function is a major cause of increased mortality and disability among ischemic stroke patients. Hence, scrutiny of the cellular and molecular mechanisms that alter BBB permeability following an ischemic injury remains of paramount importance. In this context, establishment of an in vitro model of BBB that closely simulates human cerebral barrier may offer an easy, inexpensive, and straightforward approach to identify signaling pathways involved in BBB breakdown and may help to discover new therapeutic targets to restore its damage. This chapter describes a sequential method pertaining to establishment of a triple culture model of human BBB consisting of the three main cellular components of the cerebral barrier. It also documents how the integrity and function of this barrier are evaluated through measurements of transendothelial electrical resistance (TEER) and paracellular flux of permeability marker and sodium fluorescein (NaF, 376 Da), respectively, both in normal and experimental conditions mimicking ischemic injury.
摘要：
血脑屏障（BBB），主要由脑微血管内皮细胞组成，星形胶质细胞末端脚，和周细胞,充当物理和生化屏障，选择性地限制循环分子进入脑实质。其完整性和功能的破坏是缺血性中风患者死亡率和残疾增加的主要原因。因此，对缺血性损伤后改变BBB通透性的细胞和分子机制的研究仍然至关重要.在这种情况下，建立一个密切模拟人脑屏障的体外BBB模型,便宜,和直接的方法来识别参与BBB分解的信号通路，并可能有助于发现新的治疗靶点来恢复其损伤。本章描述了与建立由脑屏障的三种主要细胞成分组成的人BBB的三重培养模型有关的顺序方法。它还记录了如何通过测量跨内皮电阻（TEER）和细胞旁通透性标记物和荧光素钠（NaF，376Da），分别,在模拟缺血性损伤的正常和实验条件下。
