Functional cardiomyocytes derived from human induced pluripotent stem cells.
Functional cardiomyocytes derived from human induced pluripotent stem cells.
复制标题
DOI：
10.1161/circresaha.108.192237
复制
发表时间：
2009-02-27
影响因子：
20.1
通讯作者：
Kamp TJ
中科院分区：
文献类型：
--
作者：
Zhang J;Wilson GF;Soerens AG;Koonce CH;Yu J;Palecek SP;Thomson JA;Kamp TJ
人类诱导多能干细胞（iPS）在心血管研究和治疗应用方面具有巨大的前景，但人类iPS细胞分化为功能性心肌细胞的能力尚未得到证实。本研究的目的是表征使用OCT 4、SOX 2、NANOG和LIN 28转基因产生的人iPS细胞与人胚胎干（ES）细胞相比的心脏分化潜力。采用胚状体（EB）法对iPS和ES细胞进行分化。虽然收缩EB的绝对百分比不同，但iPS和ES细胞系形成收缩EB的时间过程相当。iPS和ES细胞衍生的心肌细胞的RT-PCR分析显示了相似的心脏基因表达模式。多能性基因OCT 4和NANOG随着心脏分化而下调，但由于残余转基因表达，iPS细胞系中的下调被钝化。BrdU标记的iPS和ES细胞衍生的心肌细胞的增殖是相似的，分离的心肌细胞的免疫细胞化学显示难以区分的肌节组织。电生理学研究表明，iPS细胞具有像ES细胞一样的分化能力，可以根据动作电位特征分化为结节样、心房样和心室样表型。iPS和ES细胞来源的心肌细胞均表现出对β-肾上腺素能刺激的反应性，表现为自发频率增加和动作电位时程缩短。我们的结论是，人iPS细胞可以分化为功能性心肌细胞，因此iPS细胞是一个可行的选择，作为心脏修复的自体细胞来源和心血管研究的有力工具。
Human induced pluripotent stem (iPS) cells hold great promise for cardiovascular research and therapeutic applications, but the ability of human iPS cells to differentiate into functional cardiomyocytes has not yet been demonstrated. The aim of this study was to characterize the cardiac differentiation potential of human iPS cells generated using OCT4, SOX2, NANOG, and LIN28 transgenes compared to human embryonic stem (ES) cells. The iPS and ES cells were differentiated using the embryoid body (EB) method. The time course of developing contracting EBs was comparable for the iPS and ES cell lines, although the absolute percentages of contracting EBs differed. RT-PCR analyses of iPS and ES cell-derived cardiomyocytes demonstrated similar cardiac gene expression patterns. The pluripotency genes OCT4 and NANOG were downregulated with cardiac differentiation, but the downregulation was blunted in the iPS cell lines due to residual transgene expression. Proliferation of iPS and ES cell derived-cardiomyocytes based on BrdU labeling was similar, and immunocytochemistry of isolated cardiomyocytes revealed indistinguishable sarcomeric organizations. Electrophysiology studies indicated that iPS cells have a capacity like ES cells for differentiation into nodal-, atrial-, and ventricular-like phenotypes based on action potential characteristics. Both iPS and ES cell-derived cardiomyocytes exhibited responsiveness to β-adrenergic stimulation manifest by an increase in spontaneous rate and a decrease in action potential duration. We conclude that human iPS cells can differentiate into functional cardiomyocytes, and thus iPS cells are a viable option as an autologous cell source for cardiac repair and a powerful tool for cardiovascular research.