免疫磁珠体外提纯胚胎大鼠神经干细胞的实验研究

PURIFICATION AND IDENTIFICATION OF THE NEURAL STEM CELLS FROM FETAL RAT BRAIN

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 。

The objective of the experiment was to separate neural stem cells from fetal rat brain by using immunomagnetic beads, and to study their morphology and marker on cell membrane. Cell suspension of the fetal rat cerebral cortex was incubated with antibody against Nestin and the labeled cells were separated through magnetic field by immunomagnetic beads coated with the second antibody. We determined the purity of the separated cells and the cell activity by light microscopy and detected the Nestin expression ratio of the sorted cells by flow cytomety. The morphology of neural stem cells was observed with a scanning electron microscope. The results showed that the purity of the separated cells was 87 8%%+2 4%, the activity of the cells was 94 8%+2 3%, and the Nestin expression ratio of the sorted cells was 88 3%+1 04%. Binding of the sorted cells to beads was shown under scanning electron microscope.Our conclusions are the magnetic cell sorting system we used can effectively separate neural stem cells from the cerebral cortex cell suspension and maintain their activity. The sorted cells can be used in the subsequent experiments.