Leeds Institute of Cardiovascular and Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, UK.
Methods Mol Biol. 2022;2492:157-173. doi: 10.1007/978-1-0716-2289-6_9.
Constructing a reliable in vitro blood-brain barrier (BBB) model using human primary cells has been considered a major challenge during the past decades. These systems could provide valuable information regarding the effect of therapeutic compounds on different BBB cell types (endothelial cells, astrocytes, pericytes) and their ability to cross the barrier in order to reach the brain. Several attempts have been made to develop in vitro BBB models, but these studies mainly used rat, bovine, and porcine cells rather than human primary cells. Genetically modified cell lines have also been used, but they do not appear to maintain physiological properties of the BBB. Here, we describe a detailed protocol for co-culturing and maintaining human brain primary endothelial cells, pericytes, and astrocytes under flow to create an in vitro human BBB model, which can be used for toxicity testing and for studying cross-interaction among different cell types involved in the BBB formation.
在过去的几十年中，使用人原代细胞构建可靠的体外血脑屏障（BBB）模型一直是一个重大挑战。这些系统可以提供有关治疗化合物对不同 BBB 细胞类型（内皮细胞、星形胶质细胞和周细胞）的影响及其穿过屏障进入大脑的能力的有价值的信息。已经有几种尝试来开发体外 BBB 模型，但这些研究主要使用大鼠、牛和猪细胞，而不是人原代细胞。遗传修饰的细胞系也已被使用，但它们似乎无法维持 BBB 的生理特性。在这里，我们描述了一种详细的方案，用于共培养和维持在流动条件下的人脑原代内皮细胞、周细胞和星形胶质细胞，以创建体外人 BBB 模型，该模型可用于毒性测试和研究参与 BBB 形成的不同细胞类型之间的相互作用。
