刘通, 卜庆婷, 马珂, 金岩, 孙冬冬, 李秀娟, 曹丰. 心肌营养素-1诱导小鼠诱导性多能干细胞心肌细胞定向分化的实验研究[J]. 心脏杂志, 2013, 25(3): 307-310+322. DOI: 10.13191/j.chj.2013.03.61.liut.002
    引用本文: 刘通, 卜庆婷, 马珂, 金岩, 孙冬冬, 李秀娟, 曹丰. 心肌营养素-1诱导小鼠诱导性多能干细胞心肌细胞定向分化的实验研究[J]. 心脏杂志, 2013, 25(3): 307-310+322. DOI: 10.13191/j.chj.2013.03.61.liut.002
    LIU Tong, BU Qing-ting, MA Ke, JIN Yan, SUN Dong-dong, LI Xiu-juan, CAO Feng. Effects of cardiotrophin-1 on cardiac differentiation of mouse-induced pluripotent stem cells[J]. Chinese Heart Journal, 2013, 25(3): 307-310+322. DOI: 10.13191/j.chj.2013.03.61.liut.002
    Citation: LIU Tong, BU Qing-ting, MA Ke, JIN Yan, SUN Dong-dong, LI Xiu-juan, CAO Feng. Effects of cardiotrophin-1 on cardiac differentiation of mouse-induced pluripotent stem cells[J]. Chinese Heart Journal, 2013, 25(3): 307-310+322. DOI: 10.13191/j.chj.2013.03.61.liut.002

    心肌营养素-1诱导小鼠诱导性多能干细胞心肌细胞定向分化的实验研究

    Effects of cardiotrophin-1 on cardiac differentiation of mouse-induced pluripotent stem cells

    • 摘要: 目的:观察心肌营养素-1(CT-1)对小鼠诱导性多能干细胞(miPSCs)心肌细胞定向分化的诱导作用。方法:实验分为对照组与CT-1处理组。miPSCs经悬滴法诱导形成拟胚体,第3天培养基中加入1μmol/ml CT-1(CT-1组),对照组采用普通培养基。于第4、7、10和14天,收取细胞样本,采用实时PCR检测心脏特异标志物基因的表达。用免疫荧光染色及流式细胞术检测干细胞标志物Oct4和心肌特异性分化标志物肌钙蛋白I(cTnI)的表达。用透射电镜观察心肌样细胞的超微结构。结果:CT-1组第10天,样本中心肌特异性肌钙蛋白T(cTnT)基因表达的水平为同期对照组的1.75倍,有显著性差异(P<0.05)。免疫荧光染色法检测显示,两组均有cTnI表达。流式细胞术鉴定的结果显示,对照组与CT-1组cTnI的阳性率分别为28.5%和56.4%,有显著性差异(P<0.05)。电镜观察显示,CT-1组肌原纤维及胞内线粒体明显增多,肌纤维排列规则,可见细胞间桥粒连接和缝隙连接。结论:CT-1可显著提高miPSC向心肌细胞定向分化的效率。

       

      Abstract: AIM: To observe the effect of cardiotrophin-1 on cardiac differentiation of mouse-induced pluripotent stem cells(miPSCs).METHODS: Differentiation of miPS cells was initiated by embryoid body formation with the hanging drop method.In the experimental group,CT-1 application started at day 3 of the differentiation process(1 μmol/ml,CT-1 group) and normal medium was used in control group.Real-time PCR analysis,immunofluorescence staining,flow cytometry analysis and transmission electron microscope were used to examine the expression of embryonic and cardiac-specific cell markers and the cellular ultramicrostructure at days 4,7,10 and 14.RESULTS: Compared with the control group,expression of cardiac-specific marker cTnT in CT-1 group at day 10 increased 1.75 times(P<0.05).Immunofluorescence staining showed cTnI positive cells in both groups and flow cytometry analysis showed that the rates of cTnI positive cells were 28.5% and 56.4%,respectively(P<0.05).Compared with those in control group,more organized myofibril,mitochondria and cell junction formation were observed in CT-1 group.CONCULSION: CT-1 significantly promotes cardiac differentiation of miPS cells.

       

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