PMA体外诱导小鼠多能干细胞向心肌细胞的分化

发布时间：2018-07-04 23:33

本文选题：多能干细胞 + 心肌细胞　；参考：《基础医学与临床》2015年04期

【摘要】：目的研究丙二醇甲醚醋酸酯(PMA)对小鼠诱导多能干细胞体外分化为心肌细胞的影响,以建立一种高效安全的体外诱导i PSC分化为心肌细胞的实验方法。方法用悬滴法形成拟胚体(EBs),PMA诱导其向心肌细胞定向分化。免疫细胞学标记检测心肌肌钙蛋白T(c Tn T)和α横纹肌辅肌动蛋白(α-actinin)的表达;RT-PCR和q-PCR检测Brachyury,c Tn T,MLC2a,NKX2.5和GATA4等mRNA的表达。以添加相应DMSO作为对照组,观察各组出现搏动拟胚体的数量,计算分化比率。结果 PMA诱导小鼠诱导多能干细胞分化为心肌细胞的最佳浓度为100 nmol/L,此时拟胚体搏动率可达53%,显著高于对照组(12%),且PMA诱导产生的心肌细胞表达多种心肌蛋白及基因,具有心肌细胞的结构特征。结论 PMA能够促进mi PSC在体外定向分化为心肌样细胞。
[Abstract]:Objective to study the effect of propanediol methyl ether acetate (PMA) on the differentiation of mouse pluripotent stem cells into cardiomyocytes in vitro, and to establish an efficient and safe method for inducing iPSC to differentiate into cardiomyocytes in vitro. Methods the embryoid body (EBs) PMA was induced to differentiate into cardiomyocytes by suspension drop method. The expressions of cardiac troponin T (c TnT) and 伪 -striated actin (伪 -actinin) were detected by immunocytochemistry. The mRNA expressions of BrachyuryytnTnTnTnTnTnTnTnTMLC2aNKX2.5 and GATA4 were detected by RT-PCR and q-PCR respectively. The number of pulsatile embryoid bodies in each group was observed and the differentiation ratio was calculated by adding corresponding DMSO as control group. Results the optimal concentration of PMA induced pluripotent stem cells to differentiate into cardiomyocytes was 100 nmol / L, and the pulsatile rate of embryoid body was 53, which was significantly higher than that of control group (12%), and the cardiomyocytes induced by PMA expressed many kinds of myocardial proteins and genes. It has the structural characteristics of cardiomyocytes. Conclusion PMA can promote the differentiation of mi PSC into cardiomyocytes in vitro.
【作者单位】：第四军医大学唐都医院心血管内科;
【基金】：国家自然科学基金(31271039;31400832)
【分类号】：R329.2

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