Morphological and functional characterization of an in vitro blood–brain barrier model

Kathe A. Stanness、Lesnick E. Westrum、Eleonora Fornaciari、Patrizia Mascagni、Jay A. Nelson、Stephan G. Stenglein、Tim Myers、Damir Janigro

Cell culture models have been extensively used for studies of blood–brain barrier BBB.function. However, several in vitro models fail to reproduce some, if not most, of the physiological and morphological properties of in situ brain microvascular endothelial cells. We have recently developed a dynamic, tridimensional BBB model where endothelial cells exposed to intraluminal flow form a barrier to ions and proteins following prolonged co-culturing with glia. We have further characterized this cell culture model to determine whether these barrier properties were due to expression of a BBB phenotype. Endothelial cells of human, bovine or rodent origin were used. When co-cultured with glia, intraluminally grown endothelial cells developed features similar to in vivo endothelial cells, including tight junctional contacts at interdigitating processes and a high transendothelial resistance. This in vitro BBB was characterized by the expression of an abluminal, ouabain-sensitive NarK pump, and thus favored passage of potassium ions towards the lumen while preventing Kq extravasation. Similarly, the in vitro BBB prevented the passage of blood–brain barrier-impermeant drugs such as morphine, sucrose and mannitol. while allowing extraluminal accumulation of lipophylic substances such as theophylline. Finally, expression of stereo-selective transporters for Aspartate was revealed by tracer studies. We conclude that the in vitro dynamic BBB model may become an useful tool for the studies of BBB-function and for the testing of drug passage across the brain endothelial monolayer. q 1997 Elsevier Science B.V.

细胞培养模型已广泛用于血脑屏障 BBB 功能的研究。然而，一些体外模型无法重现原位脑微血管内皮细胞的某些（如果不是大部分的话）生理和形态学特性。我们最近开发了一种动态的三维 BBB 模型，其中暴露于腔内流动的内皮细胞在与神经胶质细胞长期共培养后形成离子和蛋白质的屏障。我们进一步表征了这种细胞培养模型，以确定这些屏障特性是否是由于 BBB 表型的表达。使用人、牛或啮齿动物来源的内皮细胞。当与胶质细胞共培养时，管腔内生长的内皮细胞会发展出与体内内皮细胞相似的特征，包括交叉过程中的紧密连接接触和高跨内皮阻力。这种体外 BBB 的特点是表达一种对哇巴因敏感的非管腔 NarK 泵，因此有利于钾离子向内腔通过，同时防止 Kq 外渗。同样，体外 BBB 阻止了血脑屏障不透性药物（如吗啡、蔗糖和甘露醇）的通过。同时允许亲脂性物质如茶碱的腔外积累。最后，示踪研究揭示了天冬氨酸立体选择性转运蛋白的表达。我们得出结论，体外动态 BBB 模型可能成为研究 BBB 功能和测试药物通过大脑内皮单层的有用工具。q 1997 Elsevier Science BV

文献原文：https://pubmed.ncbi.nlm.nih.gov/9401753/