A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes
A new blood-brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes
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DOI：
10.1016/j.neuint.2008.12.002
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发表时间：
2009-03-01
影响因子：
4.2
通讯作者：
Niwa, Masami
中科院分区：
文献类型：
--
作者：
Nakagawa, Shinsuke;Deli, Maria A.;Niwa, Masami
血脑屏障（BBB）的特性是由脑微血管内皮细胞和神经血管单元的相邻元件之间的串扰诱导和维持的。虽然周细胞是形态上最接近脑内皮细胞的细胞，并且共享共同的基底膜，但它们尚未用于共培养BBB模型中以测试药物渗透性。我们已经开发出一种新的同基因血脑屏障模型，其特点是使用三种主要细胞类型的脑微血管的原代培养物。内皮细胞、周细胞和星形胶质细胞的共培养模拟体内解剖学情况。在周细胞和星形胶质细胞的存在下，大鼠脑内皮细胞表达增强水平的紧密连接（TJ）蛋白occludin，claudin-5和ZO-1与典型的定位在细胞边界。通过电子显微镜提供了内皮间TJs存在的进一步形态学证据。在三重共培养中，脑内皮单层的跨内皮电阻（TEER）表明TJ的紧密性平均达到400 Ω cm（2），而荧光素的内皮渗透系数（P-e）在3 x 10（-6）cm/s的范围内。新模型中的脑内皮细胞表达葡萄糖转运蛋白-1、外排转运蛋白P-糖蛋白和多药耐药蛋白-1，并显示P-糖蛋白配体罗丹明123的极化转运。为了进一步表征模型，使用一组具有已知体内BBB渗透性的19种化合物进行药物渗透性测定。在BBB模型和体内BBB渗透性数据测量获得的体外P-e值之间发现良好的相关性（R-2 = 0.89）。新的血脑屏障模型，这是第一个模型，将周细胞在一个三重共培养设置，可以是一个有用的工具，研究血脑屏障生理学和病理学，并测试候选化合物的中枢作用的药物。(C)2008爱思唯尔有限公司版权所有。
Blood-brain barrier (BBB) characteristics are induced and maintained by cross-talk between brain microvessel endothelial cells and neighbouring elements of the neurovascular unit. While pericytes are the cells situated closest to brain endothelial cells morphologically and share a common basement membrane, they have not been used in co-culture BBB models for testing drug permeability. We have developed and characterized a new syngeneic BBB model using primary cultures of the three main cell types of cerebral microvessels. The co-culture of endothelial cells, pericytes and astrocytes mimick the anatomical situation in vivo. In the presence of both pericytes and astrocytes rat brain endothelial cells expressed enhanced levels of tight junction (TJ) proteins occludin, claudin-5 and ZO-1 with a typical localization at the cell borders. Further morphological evidence of the presence of interendothelial TJs was provided by electron microscopy. The transendothelial electrical resistance (TEER) of brain endothelial monolayers in triple co-culture, indicating the tightness of TJs reached 400 Omega cm(2) on average, while the endothelial permeability coefficients (P-e) for fluorescein was in the range of 3 x 10(-6) cm/s. Brain endothelial cells in the new model expressed glucose transporter-1, efflux transporters P-glycoprotein and multidrug resistance protein-1, and showed a polarized transport of rhodamine 123, a ligand for P-glycoprotein. To further characterize the model, drug permeability assays were performed using a set of 19 compounds with known in vivo BBB permeability. Good correlation (R-2 = 0.89) was found between in vitro P-e values obtained from measurements on the BBB model and in vivo BBB permeability data. The new BBB model, which is the first model to incorporate pericytes in a triple co-culture setting, can be a useful tool for research on BBB physiology and pathology and to test candidate compounds for centrally acting drugs. (C) 2008 Elsevier Ltd. All rights reserved.