Cardiomyocytes derived from embryonic stem cells resemble cardiomyocytes of the embryonic heart tube
Cardiomyocytes derived from embryonic stem cells resemble cardiomyocytes of the embryonic heart tube
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DOI：
10.1016/s0008-6363(03)00282-7
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发表时间：
2003-05-01
影响因子：
10.8
通讯作者：
Deprez, RHL
中科院分区：
文献类型：
--
作者：
Fijnvandraat, AC;van Ginneken, ACG;Deprez, RHL
目标：线性心管形成后，腔室特异性基因表达程序变得活跃，成为腔室心肌形成的基础。为了评估这一程序是否在体外分化的胚胎干细胞中重演，我们对体内和体外心脏发生进行了定性和定量分析。研究方法：基因表达谱由原位杂交和实时PCR和膜片钳分析心肌细胞分化HMI小鼠胚胎干细胞的时间序列和胚胎和成年小鼠心脏的电生理配置文件。结果如下：在拟胚体中，α-肌球蛋白重链、肌球蛋白轻链2a和肌内质网钙ATP酶2a的原位表达模式与心肌特异性标记基因心肌肌钙蛋白I的原位表达模式相似。肌球蛋白轻链2 v在部分心肌肌钙蛋白I表达区表达，表明心肌细胞群内存在异质性。心房利钠因子的表达，室型程序的指示，只能偶尔被检测到原位杂交。定量逆转录PCR显示，所有的心脏基因，最显着的心钠素，表达水平相对较低，类似于胚胎心脏在胚胎天8.75-9。胚胎干细胞来源的心肌细胞的电生理特性的分析表明，在体外延长分化过程中，动作电位的上升速度增加，持续时间缩短。当胚胎12.5天的胚胎小鼠心脏隔室被用作参考时，胚胎干细胞衍生的心肌细胞的相当大一部分的电生理特性最令人联想到在胚胎流出道中观察到的那些。结论：总之，这些数据表明，通过胚胎干细胞分化获得的大多数心肌细胞保持着与原代心管心肌细胞相似的表型，几乎没有任何心肌细胞形成室心肌表型。(C)2003年欧洲心脏病学会。由Elsevier Science B. V.出版，版权所有。
Objective: After formation of the linear heart tube a chamber-specific program of gene expression becomes active that underlies the formation of the chamber myocardium. To assess whether this program is recapitulated in in vitro differentiated embryonic stem cells, we performed qualitative and quantitative analyses of cardiogenesis in vivo and in vitro. Methods: Gene expression profiles were made by in situ hybridisation and real-time PCR and electrophysiological profiles by patch clamp analyses of cardiomyocytes derived from time series of differentiating HMI mouse embryonic stem cells and from embryonic and adult mouse hearts. Results: In embryoid bodies the in situ patterns of expression of alpha-myosin heavy chain, myosin light chain 2a and sarcoendoplasmic reticulum calcium ATPase 2a were similar to that of the heart muscle-specific marker gene cardiac troponin I. Myosin light chain 2v was expressed in part of the cardiac troponin I-expressing area, indicating heterogeneity within the cardiac cell population. Atrial natriuretic factor expression, indicative of the chamber-type program, could only very occasionally be detected by in situ hybridisation. Quantitative reverse transcriptase PCR showed that all cardiac genes, most notably atrial natriuretic factor, were expressed at relatively low levels, similar to those in embryonic hearts at embryonic day 8.75-9. Analysis of the electrophysiological characteristics of embryonic stem cell-derived cardiomyocytes showed an increase of the upstroke velocity and a shorter duration of the action potential during prolonged differentiation in vitro. When embryonic mouse heart compartments of embryonic day 12.5 were used as a reference, the electrophysiological characteristics of a substantial part of the embryonic stem cell-derived cardiomyocytes were most reminiscent to those observed in the embryonic outflow tract. Conclusion: Together, these data suggest that most cardiomyocytes acquired by differentiation of embryonic stem cells maintain a phenotype reminiscent of that of the cardiomyocytes of the primary heart tube, and hardly any myocytes develop a chamber myocardial phenotype. (C) 2003 European Society of Cardiology. Published by Elsevier Science B.V. All rights reserved.