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为建立免疫磁珠体外提纯胚胎神经干细胞 (NSC)的方法 ,制备胚胎SD大鼠的大脑皮质组织悬液 ,采用磁珠分选法选择Nestin阳性细胞群 ,台盼蓝染色观察活力 ,扫描电镜下行形态学观察 ,流式细胞仪检测荧光标记细胞的Nestin表达阳性率。结果发现 ,分选后所得细胞纯度达 (87 8± 2 4 ) % ,细胞活力 (94 8± 2 3) % ,流式细胞仪检测细胞Nestin阳性表达率为 (88 3± 1 0 4 ) % ,符合NSC的特点 ,扫描电镜观察可见细胞表面有数目不等的磁珠紧密结合。研究表明 ,磁性分选系统是一种有效的神经干细胞分离提纯方法 ,且对细胞活力无影响
To establish a method to purify embryonic neural stem cells (NSC) by immunomagnetic beads in vitro, the cerebral cortex tissue suspension of embryonic SD rats was prepared. Nestin positive cell population was selected by magnetic bead sorting, and the activity was observed by trypan blue staining. Morphological observation and flow cytometry were used to detect the positive rate of Nestin expression in fluorescent labeled cells. The results showed that the purity of the cells obtained after sorting was (87 8 ± 2 4)%, the cell viability was (94 8 ± 23)%, the positive expression rate of Nestin by flow cytometry was (88 3 ± 1 0 4) , In line with the characteristics of NSC, scanning electron microscopy showed a number of cells on the surface of the beads in close contact with each other. Studies have shown that magnetic separation system is an effective method of neural stem cell isolation and purification, and has no effect on cell viability