Department of Biomedical and Biotechnological Sciences, Section of Pharmacology, University of Catania, Catania, Italy.
Departement of Scienza e Tecnologia del Farmaco, Universita' di Turin, Turin, Italy.
Methods Mol Biol. 2022;2492:333-352. doi: 10.1007/978-1-0716-2289-6_20.
The blood-brain barrier (BBB) is a highly specialized structure, constituted by endothelial cells that together with astrocytes and pericytes provide a functional interface between the central nervous system and the periphery. Several pathological conditions may affect its functions, and lately BBB involvement in the pathogenesis of Alzheimer's disease has been demonstrated. Both endothelial cells and astrocytes can be differentially affected during the course of the disease. In vitro BBB models present a powerful tool in evaluating the effects that β-amyloid (Aβ), or other pathogenic stimuli, play on the BBB at cellular level. In vitro BBB models derived from human cell sources are rare and not easily implemented. We generated two conditionally immortalized human cell lines, brain microvascular endothelial cells (TY10), and astrocytes (hAST), that, when co-cultured under appropriate conditions, exhibit BBB-like characteristics. This model allowed us to evaluate the transmigration of peripheral blood mononuclear cells (PBMCs) through the in vitro barrier exposed to Aβ and the role played by astrocytes in the modulation of this phenomenon. We describe here the methodology used in our lab to set up our in vitro model of the BBB and to carry out a PBMC transmigration assay.
血脑屏障（BBB）是一种高度特化的结构，由内皮细胞与星形胶质细胞和周细胞共同构成，为中枢神经系统与外周之间提供了一个功能界面。多种病理状况可能会影响其功能，最近已证实 BBB 参与了阿尔茨海默病的发病机制。在疾病过程中，内皮细胞和星形胶质细胞可能会受到不同程度的影响。体外 BBB 模型是评估β-淀粉样蛋白（Aβ）或其他致病刺激物在细胞水平上对 BBB 影响的有力工具。源自人类细胞来源的体外 BBB 模型很少见，也不容易实施。我们生成了两种条件永生化的人类细胞系，脑微血管内皮细胞（TY10）和星形胶质细胞（hAST），当在适当的条件下共培养时，它们表现出 BBB 样特征。该模型使我们能够评估外周血单核细胞（PBMC）穿过暴露于 Aβ的体外屏障的迁移情况，以及星形胶质细胞在调节这种现象中所起的作用。我们在此描述了我们实验室用于建立体外 BBB 模型并进行 PBMC 迁移测定的方法。
