差速贴壁结合神经营养因子3体外纯化培养人胚嗅鞘细胞

目的:采用差速贴壁结合神经营养因子3的方法对人胚嗅球嗅鞘细胞进行原代纯化培养,探讨简单实用的嗅鞘细胞体外培养方法。方法:将差速贴壁后人胚嗅鞘细胞间隔48h用含100mL/L胎牛血清和含NT3的DF12培养液交替进行体外培养。观察嗅鞘细胞的生长变化,采用形态学和P75免疫细胞化学染色进行细胞及纯度鉴定。结果:体外培养的人胚嗅鞘细胞P75染色呈阳性反应,呈双极、三极细胞,细胞突起细长,并可形成细胞突起网络。在体外培养9d时可以获得95%的嗅鞘细胞,12d时为83%,且细胞状态良好。结论:差速贴壁法结合间断NT3应用是一种简单实用的嗅鞘细胞纯化培养方法。

Culture and purification of olfactory ensheathing cells from human fetal using different attachment rates combined with the technique of using NT3 intermittently

AIM: To explore a simple and pragmatic method to obtain sufficient olfactory ensheathing cells from human fetal by using different attachment rates in harvested cells with the combinating of the technique of using NT3 intermittently. METHODS: DMEM/F12 culture solution including 100 mL/L of fetal bovine serum or including NT3 was used to culture olfactory ensheathing cells intermittently every 48 h. The conditions and growth degree of OECs were observed, and P75 immunocytochemistry was used to estimate the purity of the cells. RESULTS: Human fetal OECs were positive after P75 immunocytochemistry. They appeared to be dipolar or tripolar cells and their processes formed a network in vitro. The purity of OECs in good conditions reached about 95% on 9 d and 83% on 12 d. CONCLUSION: The method of using different attachment rates combined with the thchnique of using NT3 intermittently can culture and purify OECs simply and effectively.