Evaluation of bEnd5 cell line as an in vitro model for the blood-brain barrier under normal and hypoxic/aglycemic conditions.
Evaluation of bEnd5 cell line as an in vitro model for the blood-brain barrier under normal and hypoxic/aglycemic conditions.
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DOI：
10.1002/jps.21002
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发表时间：
2007-12
影响因子：
3.8
通讯作者：
Tianzhi Yang;Karen E. Roder;T. Abbruscato
中科院分区：
文献类型：
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作者：
Tianzhi Yang;Karen E. Roder;T. Abbruscato
本研究的目的是评估小鼠内皮细胞系bEnd 5作为正常或病理（中风）条件下血脑屏障（BBB）模型的适用性。与成熟的牛脑内皮细胞（BBMEC）模型相比，培养的bEnd 5单层在第7天达到最大跨内皮电阻（TEER）121 Ω cm（2），并具有椭圆形和纺锤形形态。在结构上，bEnd 5细胞和BBMEC的汇合单层表现出紧密连接蛋白ZO-1和闭合蛋白的外周带染色。bEnd 5和BBMEC均表达重要的紧密连接蛋白ZO-1、闭合蛋白和封闭蛋白-1，以及转运蛋白P-糖蛋白（P-gp）、NKCC、GLUT 1和大多数PKC亚型。标记物渗透性实验表明，bEnd 5细胞形成了与良好建立的体外BBB模型（如BBMEC）相比的紧密屏障。在短时间缺氧/无糖血症（H/A）后，与BBMEC的较晚时间段相比，在bEnd 5内皮单层中观察到高渗透性，表明bEnd 5细胞比BBMEC对缺氧/无糖血症治疗更敏感。综上所述，bEnd 5细胞培养模型可以为药物递送研究和建模病理状态（例如与中风相关的氧葡萄糖剥夺）提供有用的血脑屏障体外模型。
The purpose of the study was to assess the suitability of the mouse endothelial cell line bEnd5 as a blood-brain barrier (BBB) model under normal or pathologic (stroke) conditions. In comparison to the well-established bovine brain endothelial cell (BBMEC) model, cultured bEnd5 monolayers reached a maximal transendothelial electrical resistance (TEER) of 121 Omega cm(2) on day 7, and possessed oval and spindle shape morphology. Structurally, confluent monolayers of bEnd5 cells and BBMECs exhibit peripheral band staining of the tight junction protein ZO-1 and occludin. Both bEnd5 and BBMECs express important tight junctional proteins, ZO-1, occludin and claudin-1, as well as the transporters P-glycoprotein (P-gp), NKCC, GLUT1, and most PKC isoforms. Marker permeability experiments suggest that bEnd5 cells form a tight barrier that compares to well-established in vitro BBB models, such as the BBMEC. After short durations of hypoxia/aglycemia (H/A), hyperpermeability was seen in the bEnd5 endothelial monolayer compared to later time periods for BBMECs, suggesting that bEnd5 cells are more sensitive to hypoxia/algycemia treatment than BBMECs. Taken together, bEnd5 cell culture model may provide a useful in vitro model of the BBB for drug delivery studies and modeling pathological states such as oxygen glucose deprivation associated with stroke.