School of Basic Medical Science, Xinxiang Medical University, Xinxiang, China/Department of Cardiovascular, the Second People's Hospital of Pingdingshan, Pingdingshan, China.
School of Basic Medical Science, Xinxiang Medical University, Xinxiang, China.
Pak J Pharm Sci. 2023 Mar;36(2):515-524.
The aim of this study was to explore the In Vitro effects of stromal-derived factor-1α (SDF-1α) on the migration and proliferation of c-kit+ cardiac stem cells. The lentivirus containing SDF-1α (LV-SDF-1α) was constructed. Primary myocardial fibroblasts were transfected by LV-SDF-1α, followed by primary culture of cardiac tissue cells and separation of c-kit+ cardiac stem cells with a flow cytometer, in order to investigate the effects of SDF-1α on the migration and proliferation of c-kit+ cardiac stem cells using cell co-culture, immunofluorescence and EdU tracing technologies. The results showed that myocardial fibroblasts could secrete SDF-1α after the transfection with LV-SDF-1α. High-purity c-kit+ cardiac stem cells were obtained through flow cytometry sorting and the positive rate was about 40%. The c-kit+ cardiac stem cells cultured In Vitro could be differentiated into cTnT positive cardiomyocyte-like cells. After co-culture of myocardial fibroblasts and c-kit+ cardiac stem cells transfected with lentivirus, SDF-1α might increase the migration of c-kit+ cardiac stem cells, but SDF-1α did not promote the proliferation of c-kit+ cardiac stem cells. In conclusion, the myocardial fibroblasts transfected with lentivirus can highly express SDF-1α, c-kit+ cardiac stem cells can be differentiated into cTnT positive cardiomyocyte-like cells and SDF-1α can effectively enhance the migration of c-kit+ cardiac stem cells but fails to stimulate the proliferation.
本研究旨在探讨基质衍生因子-1α(SDF-1α)对 c-kit+ 心脏干细胞迁移和增殖的体外影响。构建了含有 SDF-1α 的慢病毒(LV-SDF-1α)。通过 LV-SDF-1α 转染原代心肌成纤维细胞,然后进行心脏组织细胞的原代培养和流式细胞仪分离 c-kit+ 心脏干细胞,以细胞共培养、免疫荧光和 EdU 示踪技术研究 SDF-1α 对 c-kit+ 心脏干细胞迁移和增殖的影响。结果表明,LV-SDF-1α 转染后的心肌成纤维细胞可以分泌 SDF-1α。通过流式细胞术分选获得高纯度 c-kit+ 心脏干细胞,阳性率约为 40%。体外培养的 c-kit+ 心脏干细胞可分化为 cTnT 阳性心肌样细胞。心肌成纤维细胞和转染慢病毒的 c-kit+ 心脏干细胞共培养后,SDF-1α 可能增加 c-kit+ 心脏干细胞的迁移,但 SDF-1α 并未促进 c-kit+ 心脏干细胞的增殖。综上所述,转染慢病毒的心肌成纤维细胞可高表达 SDF-1α,c-kit+ 心脏干细胞可分化为 cTnT 阳性心肌样细胞,SDF-1α 可有效增强 c-kit+ 心脏干细胞的迁移,但不能刺激其增殖。