Abstract

This study aimed to demonstrate the advantage of a four-cell model of the blood–brain barrier (BBB) in vitro in comparison with the traditional three-cell model, as well as the influence of pericytes on the phenotype of endothelial cells. This paper describes a method for co-culture of primary brain microvascular endothelial cells, pericytes, astrocytes, and neurons in the Transwell BBB model in vitro. A quantitative analysis was carried out between the values of transendothelial electrical resistance (TEER), as well as between the content of endothelial cell tight junction markers in three- and four-cell Transwell models of the BBB. According to the obtained data, the presence of pericytes is accompanied by higher TEER values and a higher content of tight junction proteins. The presented results are consistent with the world scientific literature and confirm the hypothesis that pericytes not only perform a supporting function for endothelial cells, but are also an important metabolic link that regulates the barrier functions of the BBB. Thus, co-culture of cells of the neurovascular unit (NVU) of the brain with pericytes is necessary for the formation of a phenotype in endothelial cells that is close to the conditions in the NVU microenvironment in vivo.

中文翻译：

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周细胞作为 Transwell 体外血脑屏障模型中必需的细胞元素

摘要

本研究旨在论证体外血脑屏障（BBB）四细胞模型与传统三细胞模型相比的优势，以及周细胞对内皮细胞表型的影响。本文介绍了一种在Transwell BBB模型中体外共培养原代脑微血管内皮细胞、周细胞、星形胶质细胞和神经元的方法。在 BBB 的三细胞和四细胞 Transwell 模型中，对跨内皮电阻 (TEER) 值以及内皮细胞紧密连接标记物的含量进行定量分析。根据获得的数据，周细胞的存在伴随着较高的TEER值和较高含量的紧密连接蛋白。所呈现的结果与世界科学文献一致，并证实了周细胞不仅对内皮细胞发挥支持功能，而且还是调节血脑屏障屏障功能的重要代谢环节的假设。因此，