基质细胞衍生因子1趋化神经干细胞迁移的体外效应

In vitro effect of stromal cell derived factor-1 on the migration of neural stem cells

背景:神经干细胞的迁移在神经系统的发育和损伤修复中起着至关重要的作用,近来研究表明趋化因子参与神经干细胞的迁移,但关于其迁移机制目前尚不清楚。目的:观察体外条件下基质细胞衍生因子1对胎鼠海马神经干细胞的趋化迁移作用。方法:通过无血清法体外分离、培养及鉴定胎鼠海马神经干细胞;细胞免疫荧光及RT-PCR检测其CXCR4是否表达;观察不同浓度基质细胞衍生因子1对神经干细胞的趋化迁移作用,中和CXCR4受体以验证基质细胞衍生因子1趋化迁移作用的特异性。结果与结论:胎鼠海马来源神经干细胞表达趋化因子受体CXCR4,呈阳性。RT-PCR琼脂糖凝胶电泳后出现643bp特异性扩增条带。体外条件下基质细胞衍生因子1趋化迁移随浓度而增强,500μg/L为最佳趋化浓度。加入抗CXCR4多克隆抗体中和后,神经干细胞迁移较基质细胞衍生因子1组明显减少,与对照组比较,差异无显著性意义(P>0.05),提示抗CXCR4多克隆抗体可阻断趋化迁移作用。

BACKGROUND：Neural stem cell migration plays important roles in the development and repair of the central nervous system.Although recent studies have shown that the chemokines mediate neural stem cell migration,but the mechanism is unclear.OBJECTIVE：To explore the effects of stromal cell-derived factor-1（SDF-1） on migration of fetal rat hippocampus neural stem cells in vitro.METHODS：The fetal rat hippocampal neural stem cells were isolated,cultured and identified in serum-free medium.The expression of CXCR4 in neural stem cells was detected using immunofluorescence and RT-PCR.The under-agarose cell migration assay was used to observe the effects of SDF-1（50-500 ng/ml） on neural stem cell migration,and blocking CXCR4,the receptor of SDF-1,to identify the specificity of migration.RESULTS AND CONCLUSION：Immunofluorescence results showed that neural stem cells were CXCR4 positive,and RT-PCR findings showed 643 bp specific band in agarose gel electrophoresis.The Under-Agarose cell migration assay showed that：SDF-1（50-500 ng/ml） could accelerate the migration of neural stem cells,the migration increased with the concentration,and 500 μg/L was the best.By adding anti-CXCR4 polyclonal antibodies,the migration of neural stem cells compared with SDF-1 was significantly reduced,no significant difference with the control group（P 〉0.05）,pointing out anti-CXCR4 polyclonal antibodies can block the effect of the migration.