骨髓干细胞体外培养诱导为心肌样细胞的研究  

Differentiation of human mesenchymal stem cells into cardiomyocytes

作　　者：佟铁壁[1] 孙旗[1] 胡煜[2] 

机构地区：[1]解放军第202医院,沈阳市110003 [2]东方医疗集团箐华医院,沈阳市110001

出　　处：《实用医学杂志》2006年第19期2219-2222,共4页The Journal of Practical Medicine

基　　金：辽宁省科学技术计划立项课题(编号:2002225002-3)

摘　　要：目的:探讨应用5-氮胞苷(5-aza)诱导体外培养人骨髓干细胞向心肌细胞分化的作用。方法:采用正常志愿者髂后上棘为穿刺点获得骨髓干细胞,体外培养传3代后应用5×10-6mmol/L5-aza(Sigma公司)诱导20h后继续原条件培养,相差显微镜下观察细胞形态并照相记录。诱导后细胞行a-actionTrophoninT免疫组化染色。行透射电镜观察,进一步行RT-PCR分析诱导后细胞表达a-cardiacaction,CardiactropponinT等mRNA。结果:诱导组人a-actionTrophoninT免疫组化染色阳性,电镜下观察到细胞与细胞间见类心肌闰盘样结构,见线粒体空泡及微丝样结构,PCR结果发现有a-cardiacaction、CardiactropponinTmRNA的高表达。结论:初步证实人骨髓干细胞在体外被5-aza诱导时,能够向心肌细胞分化。Objective To explore 5-azacitidine-induced differentiation of human bonemesenchymal stem cells in vitro into cardiomyocytes. Methods Mesenchymal stem cells(MSCs) from healthy volunteers were cultured in vitro and then subcultured for three generations. They were then induced by 5-azacytidine 5×10-6 mmol/L for 20 hours and continued to be cultured under the previous condition. The cellular morphology was observed by phasecontrast microscopy.The differentiated cells in immunohistochemical staining with a-action Trophonin T were observed under atransmission electron microscope. mRNA expressions of a-cardiac action and Cardiac tropponin T in those cells were further detected by RT-PCR. Results Positive staining with a-action Trophonin T was found in the differentiated cells.Intercalated disk-like structure between the cells together with mitochondria vacuole and microfilament-like structure in the cytoplasm was found by transmission electron microscopy. High mRNA expression of a-cardiac action and Cardiac tropponin T was found by PCR. Conclusion It is preliminarily confirmed that human MSCs in vitro can be induced by 5-azacytidine into cardiomyocytes.

关 键 词：干细胞 诱导分化 心肌样细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R542.22[医药卫生—基础医学]