新生大鼠雪旺细胞的体外培养和纯化

目的:探索简单有效获取大量高纯度雪旺细胞的新方法。方法:分离出生3d的新生SD大鼠坐骨神经,使用胰蛋白酶及I型胶原酶混合消化,结合差速贴壁法和G418纯化。四氮唑盐(MTT)比色法测定细胞的生长增殖情况,免疫细胞化学方法计算雪旺细胞的纯度。结果:胰酶和胶原酶混合消化后的活细胞率为94.6±3.4%,纯化后的雪旺细胞纯度为95.6±2.5%。结论:胰酶和胶原酶混合消化结合差速贴壁和G418纯化是获取大量高纯度雪旺细胞的简单有效方法。

Culture and Purification of Schwann Cells from Newborn Rats

Objective To explore a novel feasible and effective way to obtain Schwann cells massively and purely.Methods The sciatic nerves were isolated from 3-day-old SD rats and digested with collagenase and trypsin,Schwann cells were purified with 30-min difference adhesion 2 times and G418 selection.The growth and proliferation of the cells was estimated depending on MTT assay.Its purity was calculated by immunocytochemical staining and microphotography techniques.Results The viability was 94.6±3.4%,the purity of Schwann cells was 95.6±2.5%.Conclusion Digested with mixed enzyme and purified with 30-min difference adhesion 2 times and G418 selection is an easy,effective way to obtain massive and highly purified Schwann cells.