Differentiation of human embryonic stem cells to cardiomyocytes - Role of coculture with visceral endoderm-like cells
Differentiation of human embryonic stem cells to cardiomyocytes - Role of coculture with visceral endoderm-like cells
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DOI：
10.1161/01.cir.0000068356.38592.68
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发表时间：
2003-06-03
期刊：
影响因子：
37.8
通讯作者：
Tertoolen, L
中科院分区：
文献类型：
--
作者：
Mummery, C;Ward-van Oostwaard, D;Tertoolen, L
背景-来源于人胚胎干细胞的心肌细胞可用于心肌梗死或心力衰竭后心脏功能的恢复。在这里，我们诱导心肌细胞分化的hES细胞的一种新的方法，并比较其电生理特性和耦合与原代人胎儿myocardialcytes.Methods和结果- hES细胞与内脏内胚层（VE）样细胞从小鼠共培养。这引发了向搏动肌的分化。肌节标记蛋白、变时性反应和离子通道表达和功能是心肌细胞的典型特征。电生理结果表明，大多数细胞类似于人胎儿心室细胞。实时细胞内钙测量，荧光黄注射，和连接蛋白43表达表明，胎儿和hES衍生的心肌细胞在培养中通过间隙连接偶联。维拉帕米的电反应的抑制证明了功能性α（1C）-钙离子channels.Conclusions的存在-这是第一次证明诱导心肌细胞分化的hES细胞，不进行自发性心脏发生。它为研究培养的人类心肌细胞提供了一个模型，并可能在心肌细胞移植治疗的发展中向前迈出一步。
Background - Cardiomyocytes derived from human embryonic stem (hES) cells could be useful in restoring heart function after myocardial infarction or in heart failure. Here, we induced cardiomyocyte differentiation of hES cells by a novel method and compared their electrophysiological properties and coupling with those of primary human fetal cardiomyocytes.Methods and Results - hES cells were cocultured with visceral-endoderm (VE) - like cells from the mouse. This initiated differentiation to beating muscle. Sarcomeric marker proteins, chronotropic responses, and ion channel expression and function were typical of cardiomyocytes. Electrophysiology demonstrated that most cells resembled human fetal ventricular cells. Real-time intracellular calcium measurements, Lucifer yellow injection, and connexin 43 expression demonstrated that fetal and hES-derived cardiomyocytes are coupled by gap junctions in culture. Inhibition of electrical responses by verapamil demonstrated the presence of functional alpha(1c)-calcium ion channels.Conclusions - This is the first demonstration of induction of cardiomyocyte differentiation in hES cells that do not undergo spontaneous cardiogenesis. It provides a model for the study of human cardiomyocytes in culture and could be a step forward in the development of cardiomyocyte transplantation therapies.