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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。
AIM: To investigate the mechanism of stromal cell-derived factor-1 (SDF-1) enhancing the proliferation and migration of mouse cardiac stem cells (CSCs). Methods: CSCs were isolated from mouse myocardium and c-kit (+) CSCs were screened by immunomagnetic beads method. The surface markers of CSCs were detected by flow cytometry: c-kit of stem cell factor, Sca-1 . The c-kit (+) CSCs were treated with SDF-1 and SDF-1 specific antagonist AMD3100, the mRNA and protein expressions of c-kit were detected by qPCR and Western blotting, and the cell proliferation was detected by CCK-8 kit and transwell chemotaxis assay And migration ability. Results: After isolated and cultured cells were screened by immunomagnetic beads, the expression of c-kit and Sca-1 were all detected by flow cytometry in more than 80%. After the intervention of SDF-1, the expression of c-kit mRNA and protein were increased by qPCR and Western blotting. The chemotaxis assay of CCK-8 and transwell showed that the proliferation and migration ability of SDF-1 cells were significantly increased after intervention with SDF-1, Antagonist. Conclusion: SDF-1 enhances the proliferation and migration of c-kit (+) CSCs by promoting c-kit expression.