免疫磁珠法分离纯化SD大鼠神经干细胞

Immunomagnetic indirect positive sorting of neural stem cells from SD rat brain

目的应用微小免疫磁珠分离提纯SD胎鼠大脑皮层组织中的神经干细胞并进行培养,为研究神经干细胞的特性与神经干细胞的培养和移植研究创造有利条件。方法取SD胎鼠的大脑皮层组织制成单细胞悬液,用微小磁珠(平均直径≤50 nm)分选出神经巢蛋白(nestin)阳性的细胞群,以流式细胞术检测阳性细胞纯度,以台盼蓝染色法检测细胞活性。结果分离的神经干细胞流式细胞仪检测细胞nestin阳性表达率为(88.5±3.6)%,细胞存活率为(96.3±1.8)%。结论微小免疫磁珠法分离胎鼠脑神经干细胞群落简便、有效,可以为神经干细胞的细胞培养和移植提供细胞来源。

Objective To isolate the neural stem cells from the fetal rat brain by immunomagnetic beads technique（d ≤ 50 nm）, and then to culture purified cells for studying the property of these ceils. Methods Cell suspension of the fetal rat cerebral cortex was incubated with antibody of nestin and these labeled ceils were separated from the suspension through the magnetic field by immunomagnetic beads coated with the second antibody. Purity of the sorted cells was determined with flow cytometry. Results Purity of the sorted neural stem cells were determined as （88.5 ± 3.6）%, viable cells account for （96.3 ± 1.8） %. Conclusion The magnetic cell sorting system can separate neural stem cells effectively from brain cell suspension.