不同纯化方法体外培养人胚嗅球嗅鞘细胞

目的 采用不同纯化方法原代培养人胚嗅球嗅鞘细胞(OECs),探讨简单实用可获得高纯度人胚嗅球OECs的体外培养方法.方法 以差速贴壁法为基础结合阿糖胞苷法、无血清饥饿法和问断神经营养因子3(NT3)法纯化培养人胚嗅球OECs.观察不同纯化方法下OECs生长变化,采用免疫细胞化学染色进行纯度鉴定.结果 体外培养的人胚嗅球OECs为双极、三极细胞.细胞突起细长,并可形成细胞突起网络.差速贴壁法培养3～6 d时纯度约为88%,以后成纤维细胞增殖,OECs纯度迅速下降.无血清饥饿法培养3～6 d时OECs纯度约为90%,但细胞状态差.间断NT3法培养3～9 d时OECs纯度约为95%,且细胞状态良好.经统计学处理,第6 d后差速贴壁 间断应用NT2法优于其他方法,细胞纯度的差异有统计学意义(P<0.05).结论 差速贴壁结合同断NT3法可获得高纯度状态良好的OECs,是一种简单实用的OECs纯化培养方法 .

Culture and Purification of Human Fetal OB Olfactory Ensheathing Cells in vitro with Different Methods

Objective To explore simple and useful methods for primary culture of olfactory ensheathing cells(OECs),which could harvest human fetal OB olfactory ensheathing cells(OB-hOECs) with high purity in vitro.Methods OB-hOECs were cultured and purified with different methods.The basic method was dependent on the differing rates of cell attachment,the other methods were combined the basic method with cytarabine method,starvation method without serum,and NT3 method without serum respectively.The growth of OECs was observed and the purity of cultured cells was identified with immunocytochemistry.Results OB-hOECs appears dipolar and tripolar shape,and have slender mess evection forming network.The purity of OECs reached 88% with the basic method in 6 div,then reduced quickly due to fibroblast proliferation.With the starvation method,the OECs reached a higher purity of 91% at 6 div and 86% at 9 div,but cell condition was not very well.For the method of NT3 without serum,the purity could touch the highest score 95% at 9 div,and 83% at 12 div with a good cell condition.The purity of OECs cultured by the method of NT3 without serum is statistical significantly higher than that of other methods(P<0.05).Conclusion Associated with the basic method of different cell attachment rate,the NT3 without serum method could obtain OECs with higher purity and better cell condition.