基质细胞衍生因子-1通过促进c-kit表达提高小鼠心肌干细胞增殖和迁移能力的研究-东南大学学报医学版

基质细胞衍生因子-1通过促进c-kit表达提高小鼠心肌干细胞增殖和迁移能力的研究

单位：1. 东南大学医学院, 江苏南京 210009;
2. 东南大学附属中大医院心内科, 江苏南京 210009

分类号：Q7

出版年·卷·期（页码）：2013·32·第六期（674-679）

摘要：

目的：探讨基质细胞衍生因子-1（SDF-1）提高小鼠心肌干细胞（CSCs）增殖和迁移能力的机制。方法：从小鼠心肌中分离、培养出CSCs，并用免疫磁珠法筛选出c-kit（+）CSCs，用流式细胞仪检测CSCs表面标志：干细胞因子受体c-kit、干细胞抗原Sca-1。用SDF-1及SDF-1特异性拮抗剂AMD3100干预c-kit（+）CSCs，qPCR及Western blotting检测c-kit的mRNA和蛋白表达，用CCK-8试剂盒及transwell趋化实验检测细胞增殖和迁移能力。结果：分离、培养出的细胞经免疫磁珠筛选后，流式细胞仪检测c-kit和Sca-1表达均大于80%。SDF-1干预后，qPCR和Western blotting结果显示c-kit mRNA和蛋白表达均增高，CCK-8及transwell趋化实验结果显示SDF-1干预后细胞的增殖及迁移能力明显提高，并且可以被AMD3100拮抗。结论：SDF-1通过促进c-kit表达提高c-kit（+）CSCs的增殖及迁移能力。

Objective: To investigate the mechanism on improving proliferation and migration abilities of cardiac stem cells (CSCs) by stromal cell derived factor-1 (SDF-1). Methods: CSCs were isolated from adult mice hearts purified by magnetic-activated c-kit cell sorting magnetic beads. The markers of c-kit and Sca-1 were measured by FACS. The cells were cultured with SDF-1 and CXCR4-selective antagonist AMD3100, and c-kit expression was measured by qPCR and Western blotting. After the intervention of SDF-1 and AMD3100, proliferation and migration of c-kit(+)CSCs were measured by CCK-8 assay and transwell assay. Results: More than 80% cardiosphere-derived cells which were purified by magnetic-activated c-kit cell sorting magnetic beads were positive for c-kit and Sca-1 by FACS. SDF-1 could enhance the expression of c-kit mRNA and protein, and AMD3100 could inhibit this function. And SDF-1 could enhance the proliferation and migration of c-kit(+)CSCs, and AMD3100 could inhibit these functions. Conclusion: SDF-1 can improve the proliferation and migration abilities of CSCs by SDF-1.

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