Isobe I, Watanabe T, Yotsuyanagi T, Hazemoto N, Yamagata K, Ueki T, Nakanishi K, Asai K, Kato T
Department of Bioregulation Research, Nagoya City University Medical School, Japan.
Neurochem Int. 1996 May-Jun;28(5-6):523-33. doi: 10.1016/0197-0186(95)00142-5.
Astrocytic contribution of endothelial cell monolayer permeability was examined in two blood-brain barrier (BBB) models, using the coculture in a double chamber system: rat astrocytes and bovine aortic endothelial cells (BAECs) or bovine brain endothelial cells (BBECs). In system 1, where astrocytes were separated from endothelial cells, a 40% reduction in L-glucose permeability of the BBEC monolayer, but not the BAEC monolayer, was observed by cocultivation with astrocytes. Although several passages of BBEC in culture elicited morphological transformation from spindle-shapes to cobblestone-like features, the passaged BBECs remained responsive to astrocytes in coculture in system 1 (37% reduction of the L-glucose permeability). By contrast, in system 2, where respective endothelial cells and astrocytes layered on the upper and lower surfaces of a membrane, the permeability of both BAEC and BBEC monolayers was reduced by cocultivation with astrocytes (75% reduction for BAEC and 40% reduction for BBEC). BAECs in this contiguous coculture (system 2) with astrocytes showed numerous tight junction-like structures characteristic of the BBB in vivo. These results suggest that primary cultured BBECs, which had been primed by astrocytes in vivo, retain a higher sensitivity to astrocytes possibly through an astrocytic soluble factor (s) to exhibit BBB-specific phenotypes, and that even BAEC from extra-neural tissues, when cultured with astrocytes in close proximity in vitro, may acquire the similar phenotypes and serve for an extensive use of BBB model in vitro.
在两种血脑屏障（BBB）模型中，利用双室系统中的共培养，研究了星形胶质细胞对内皮细胞单层通透性的影响：大鼠星形胶质细胞与牛主动脉内皮细胞（BAECs）或牛脑内皮细胞（BBECs）。在系统1中，星形胶质细胞与内皮细胞分离，通过与星形胶质细胞共培养，观察到BBEC单层的L-葡萄糖通透性降低了40%，而BAEC单层未出现这种情况。尽管培养的BBEC经过几次传代后形态从纺锤形转变为鹅卵石样，但传代后的BBEC在系统1的共培养中对星形胶质细胞仍有反应（L-葡萄糖通透性降低37%）。相比之下，在系统2中，内皮细胞和星形胶质细胞分别位于膜的上表面和下表面，与星形胶质细胞共培养可降低BAEC和BBEC单层的通透性（BAEC降低75%，BBEC降低40%）。在与星形胶质细胞的这种连续共培养（系统2）中，BAEC显示出许多体内BBB特有的紧密连接样结构。这些结果表明，在体内已由星形胶质细胞预处理的原代培养BBEC，可能通过一种星形胶质细胞可溶性因子，对星形胶质细胞保持更高的敏感性，以表现出BBB特异性表型，而且即使是来自神经外组织的BAEC，当在体外与星形胶质细胞近距离共培养时，也可能获得类似的表型，从而广泛应用于体外BBB模型。
