乳鼠雪旺氏细胞的纯化培养研究

Purification of newborn rat Schwann cells in vitro

目的：获得高纯度的乳鼠雪旺氏细胞。方法：采用出生后３～５天乳鼠的坐骨、臂丛神经，植块法培养乳鼠雪旺氏细胞；并通过精细剥除神经外膜、组织块反复再植、低浓度胰酶快速消化、差速贴壁法的有机结合对雪旺氏细胞进行纯化；继用抗Ｓ－１００蛋白单抗通过间接免疫荧光法及ＡＢＣ法进行细胞鉴定。结果：首次植块获得的细胞经纯化后雪旺氏细胞可高达８５％以上，反复植块者可达９５％，甚至高达９８％。结论：本方法简单、稳定，所获得的雪旺氏细胞纯度高、数量大。

Objective:To acquire the highly purified population of Schwann cells from the newborn rats.Methods:Three to five day old SD mice were sacrificed by decapitation and their sciatic nerves and brachial plexus nerves were removed aseptically.Then the cultured and purified Schwann cells were carefully stripped off the nerve epineurium and perineurium,and repeated explantation,rapid trypsinization and differential adhesion methods were followed.The cells were identified with anti S100 protein antibody by indirect immunofluoresent methods and ABC methods.Results:The percentage of Schwann cells purified from the first explantation was over 85%,the repeated explantation′s was over 95%,even more than 98%.Conclusions:The methods are stable,simple and convenient,and the Schwann cells are high in purification,large in quantity and little affected by non experimental factors.