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目的对脑微血管内皮细胞分离、培养方法进行探索性研究,并对目前常用的纯化方法进行比较。方法首先采用两次酶消化、两次梯度离心法对大鼠脑血管内皮原代细胞进行分离,然后分别采用免疫磁珠和Thy1.1抗体杀伤法分别进行纯化。结果采用两次酶消化、两次梯度离心法可以成功分离到脑微血管内皮细胞,磁珠纯化法可得到纯度高但是数量较少的细胞,Thy1.1抗体杀伤法可得到纯度及得率均较高的内皮细胞。结论我们所摸索的方法能成功进行纯度较高的大鼠脑微血管内皮细胞原代培养。
Objective To explore the methods of isolation and culture of brain microvascular endothelial cells and to compare the purification methods commonly used at present. Methods Two primary enzymatic digestion and gradient centrifugation were used to separate primary cells of rat cerebral vascular endothelium and purified by immunomagnetic beads and Th1.1 antibody respectively. The results of two enzymatic digestion, two gradient centrifugation can be successfully isolated brain microvascular endothelial cells, the purification method of beads can be obtained with high purity but a small number of cells, Thy1.1 antibody killing method available purity and yield were High endothelial cells. Conclusion The method we have explored can successfully perform primary culture of rat brain microvascular endothelial cells with high purity.