大鼠骨髓间充质干细胞体外诱导分化为心肌样细胞的研究  

Study of in Vitro Induction of Rat Bone Marrow Mesenchymal Stem Cells into Cardiomyocyte-like Cells

作　　者：顾健[1] 王爱玲[1] 郝玉瑜[1] 张梦达[1] 孙继敏[1] 

机构地区：[1]安徽医科大学第一附属医院心血管内科,合肥230022

出　　处：《医学综述》2014年第6期1109-1111,1116,F0003,共5页Medical Recapitulate

摘　　要：目的 观察体外不同诱导条件下大鼠骨髓间充质干细胞（BMSCs）在不同时间段分化为心肌样细胞的能力及表达心肌细胞特异性标志物的差别.方法 分离培养SD大鼠BMSCs和SD乳鼠心肌细胞并鉴定,取第3代BMSCs分别进行5-氮胞苷（5-Aza）诱导、与心肌细胞共培养及单独培养,分别于第2、4、6、8、10周使用倒置显徼镜观察细胞生长情况和形态变化,反转录-聚合酶链反应检测每组细胞心肌特异性基因肌钙蛋白I（cTnI）、缝隙连接蛋白43（Cx43）和心肌特异性转录因子4（GATA-4）的表达.结果 对照组未见有搏动细胞,cTnI、Cx43和GATA-4基因表达阴性;5-氮胞苷诱导组于第2周时细胞呈梭形且排列方向趋于一致,未见有搏动细胞,cTnI、Cx43基因表达第8周达到高峰,GATA-4基因表达第4周达高峰;共培养组细胞多为梭形,呈肌性排列,可见搏动频率不一致的搏动细胞,cTnI、Cx43基因表达于第2～8周表达逐渐增加,GATA-4基因表达第6周达高峰;相同时段内,共培养组的cTnI、Cx43和GATA-4基因表达高于5-氮胞苷诱导组（P＜0.05）.结论 BMSCs经5-氮胞苷诱导或与心肌细胞共培养可转化为心肌样细胞并表达基因cTnI、Cx43和GATA-4,且共培养组BMSCs分化为心肌样细胞的能力比5-氮胞苷诱导组强,推测心肌微环境可促进BMSCs定向分化为心肌样细胞.Objective To study the ability of rat bone marrow mesenchymal stem cells（BMSCs） differentiating into cardiomyocyte-like cells and differences in expression of specific markets of myocardial cells under different in vitro inducing conditions in different periods.Methods SD rat BMSCs,SD neonatal rat cardiomyocytes were isolated,cultured and identified.The 3rd generation of BMSCs was induced respectively by 5-azacytidine （5-Aza）,and co-cultured with neonatal rat cardiomyocytes and cultured alone.An inverted microscope was used to observe cell growth and morphological changes at 2,4,6,8,10 weeks,and each set of cardiac-specific genes of cTnI,Cx43 and cardiac-specific GATA-4 was detected by RT-PCR.Results In the control group,there was no beating cells,cTnI,Cx43 and GATA-4 gene expression were negative;in 5-Aza induced cell group,cells were fusiform and arranged in the direction consistent in the first two weeks,and no beating cells.cTnI,Cx43 gene expression reached the peak at the eighty weeksGATA-4 gene expression reached the peak at the fourth week ; the co-cultured cells were mostly fusiform,in muscular arrangement,the inconsistent frequency pulsating throb cells were observed,cTnI,Cx43 gene expression gradually increased during the 2-8 weeks,GATA-4 gene expression reached peak at the sixth week.The expression of GATA-4,cTnI and Cx43 gene were higher in the co-culture group than the 5-Aza induced group during the same time period （P 〈 0.05）.Conclusion BMSCs induced by 5-Aza or co-cultured with neonatal rat cardiomyocytes can be transformed into cardiomyocyte-like cells and express cTnl,Cx43 and GATA-4 gene,and the BMSCs co-cultured cells group has more ability to differentiate into cardiomyocyte-like cells than 5-Aza induced group.It＇s speculated that myocardial microenvironment may promote BMSCs to differentiate into cardiomyocyte-like cells.

关 键 词：大鼠骨髓间充质干细胞 乳鼠心肌细胞 5-氮胞苷 共培养 诱导分化 

分 类 号：R541.4[医药卫生—心血管疾病]