大鼠脑微血管内皮细胞与星形胶质细胞共培养血脑屏障体外模型的建立

Establishment of An in vitro BBB Model with Co-cultured Rat Brain Microvessel Endothelial Cells and Astrocytes

关键词:  血脑屏障  共培养技术  体外模型  脑微血管内皮细胞  星形胶质细胞

Key words:  brain microvessel endothelial cells (BMEC)  astrocyte (AC)  co-culture  in vitro blood-brain barrier

摘要:

[目的]建立大鼠脑微血管内皮细胞与星形胶质细胞共培养血脑屏障体外模型。[方法]采用原代培养脑微血管内皮细胞与星形胶质细胞共培养方法建立血脑屏障体外模型。[结果]星形胶质细胞与脑微血管内皮细胞共培养可提高血脑屏障特异性酶——γ-谷胺酰胺转肽酶、碱性磷酸酶的表达,提高脑微血管内皮细胞跨细胞间电阻。[结论]成功建立脑微血管内皮细胞与星形胶质细胞共培养血脑屏障体外模型,而且较单层内皮细胞模型更接近在体状态。

Abstract:

[Objective] To establish an in vitro BBB model with co-cultured rat brain microvessel endothelial cells and astrocytes. [Methods] Rat brain microvessel endothelial cells (BMEC) and astrocytes (AC) were isolated and purified respectively and in vitro BBB model was established with a co-cultured method of BMEC and AC. The BMEC was inoculated on the inferior surface allowing their endfeet to contact with the basal surface of the endothelium or its basal lumina. Transendothelial electrical resistance (TEER) γ-glutamyl transpeptidase (γ-GT) and alkaline hosphatase (ALP) activity of the BBB immunohistochemistry staining of tight junction protein (ZO-1) were served as the evaluating criterion of the BBB model. [Results] Co-culture of astrocytes and brain microvessel endothelial cells could enhance the expression of the BBB-special enzymes. It could also increase the transendothelial electrical resistance. [Conclusion] The in vitro BBB model was established and can be used in the drug research.