Biegel D, Spencer D D, Pachter J S
Department of Physiology, University of Connecticut Health Center, Farmington 06030, USA.
Brain Res. 1995 Sep 18;692(1-2):183-9. doi: 10.1016/0006-8993(95)00511-n.
A simplified protocol for isolating brain microvessel endothelial cells (BMEC) from human cortex and culturing them on a thick collagen plug is described. This method results in the establishment of monolayers of BMEC that retain numerous properties indicative of the blood-brain barrier (BBB) phenotype, such as elevated transendothelial electrical resistance, attenuated paracellular flux of sucrose, peripheral actin filament distribution and asymmetric localization of the efflux peptide, P-glycoprotein, to the apical (luminal) BMEC surface. The novel 3-dimensional nature of this model system renders it ideally suitable for assaying such varied aspects of BBB physiology as solute transport, pathogen penetrance, leukocyte infiltration and tumor metastasis into the brain. Moreover, the fact that the system is derived from human brain allows for the study of pathogenetic mechanisms that may only be operative in humans.
本文描述了一种从人皮质中分离脑微血管内皮细胞（BMEC）并将其培养在厚胶原塞上的简化方案。该方法可建立BMEC单层，其保留了许多指示血脑屏障（BBB）表型的特性，如跨内皮电阻升高、蔗糖细胞旁通量减弱、外周肌动蛋白丝分布以及外排肽P-糖蛋白在BMEC顶端（管腔）表面的不对称定位。该模型系统的新型三维性质使其非常适合用于分析BBB生理学的各种方面，如溶质转运、病原体穿透、白细胞浸润和肿瘤向脑内转移。此外，该系统源自人脑这一事实使得能够研究可能仅在人类中起作用的致病机制。
