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目的分离、培养原代脑微血管内皮细胞,建立体外血脑屏障模型。同时探索高纯度、活性好的脑微血管内皮细胞的分离培养方法。方法取3周大鼠大脑,分离皮质,经过匀浆、葡聚糖离心以及消化后,取纯度较高的脑微血管段种植于胶原蛋白包被过的塑料培养瓶中进行培养。显微镜观察及检测Ⅷ因子相关抗原。结果镜下细胞呈长梭形,7d左右细胞可融合,Ⅷ因子相关抗原免疫组化检测为阳性,且阳性细胞占绝大部分。结论本实验成功分离大鼠脑微血管内皮细胞,并进行原代培养,为脑微血管内皮细胞的进一步研究提供了模型,也为构建更高级的大鼠体外血脑屏障奠定基础。
Objective To isolate and culture primary brain microvascular endothelial cells and establish an in vitro model of blood-brain barrier. At the same time to explore high purity, activity of cerebral microvascular endothelial cell isolation and culture methods. Methods The brain of rats was taken for 3 weeks and the cortex was isolated. After homogenization, dextran centrifugation and digestion, the brain microvessels with high purity were planted in collagen-coated plastic culture flasks for culture. Microscopic examination and detection of factor Ⅷ related antigen. Results The cells were long fusiform, and the cells could be fused on the 7th day. Immunohistochemical staining of factor Ⅷ related antigen was positive, and the positive cells accounted for the vast majority. Conclusion The experiment successfully isolated and cultured rat brain microvascular endothelial cells, which provided a model for the further study of brain microvascular endothelial cells and laid the foundation for the construction of higher blood-brain barrier in vitro.