东南大学学报（医学版）
東南大學學報（醫學版）
동남대학학보（의학판）
JOURNAL OF SOUTHEAST UNIVERSITY(MEDICAL SCIENCE EDITION)
2013年
6期
674-679
,共6页
陈中璞%潘啸东%姚玉宇%马根山
陳中璞%潘嘯東%姚玉宇%馬根山
진중박%반소동%요옥우%마근산
c-kit%心肌干细胞%基质细胞衍生因子-1%CXCR4%小鼠
c-kit%心肌榦細胞%基質細胞衍生因子-1%CXCR4%小鼠
c-kit%심기간세포%기질세포연생인자-1%CXCR4%소서
c-kit%cardiac stem cells%stromal cell derived factor-1%CXC chemokine receptor 4%mice
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。
目的：探討基質細胞衍生因子-1（ SDF-1）提高小鼠心肌榦細胞（ CSCs ）增殖和遷移能力的機製。方法：從小鼠心肌中分離、培養齣CSCs ，併用免疫磁珠法篩選齣c-kit （＋） CSCs ，用流式細胞儀檢測CSCs錶麵標誌：榦細胞因子受體c-kit、榦細胞抗原Sca-1。用SDF-1及SDF-1特異性拮抗劑AMD3100榦預c-kit（＋） CSCs，qPCR及Western blotting檢測c-kit的mRNA和蛋白錶達，用CCK-8試劑盒及transwell趨化實驗檢測細胞增殖和遷移能力。結果：分離、培養齣的細胞經免疫磁珠篩選後，流式細胞儀檢測c-kit和Sca-1錶達均大于80％。 SDF-1榦預後， qPCR和Western blotting 結果顯示c-kit mRNA 和蛋白錶達均增高， CCK-8及transwell趨化實驗結果顯示SDF-1榦預後細胞的增殖及遷移能力明顯提高，併且可以被AMD3100拮抗。結論：SDF-1通過促進c-kit錶達提高c-kit（＋） CSCs的增殖及遷移能力。
목적：탐토기질세포연생인자-1（ SDF-1）제고소서심기간세포（ CSCs ）증식화천이능력적궤제。방법：종소서심기중분리、배양출CSCs ，병용면역자주법사선출c-kit （＋） CSCs ，용류식세포의검측CSCs표면표지：간세포인자수체c-kit、간세포항원Sca-1。용SDF-1급SDF-1특이성길항제AMD3100간예c-kit（＋） CSCs，qPCR급Western blotting검측c-kit적mRNA화단백표체，용CCK-8시제합급transwell추화실험검측세포증식화천이능력。결과：분리、배양출적세포경면역자주사선후，류식세포의검측c-kit화Sca-1표체균대우80％。 SDF-1간예후， qPCR화Western blotting 결과현시c-kit mRNA 화단백표체균증고， CCK-8급transwell추화실험결과현시SDF-1간예후세포적증식급천이능력명현제고，병차가이피AMD3100길항。결론：SDF-1통과촉진c-kit표체제고c-kit（＋） CSCs적증식급천이능력。
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 .
