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 alpha1c-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.
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 alpha1c-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.
Authors: David A Elliott; Stefan R Braam; Katerina Koutsis; Elizabeth S Ng; Robert Jenny; Ebba L Lagerqvist; Christine Biben; Tanya Hatzistavrou; Claire E Hirst; Qing C Yu; Rhys J P Skelton; Dorien Ward-van Oostwaard; Sue Mei Lim; Ouda Khammy; Xueling Li; Susan M Hawes; Richard P Davis; Adam L Goulburn; Robert Passier; Owen W J Prall; John M Haynes; Colin W Pouton; David M Kaye; Christine L Mummery; Andrew G Elefanty; Edouard G Stanley Journal: Nat Methods Date: 2011-10-23 Impact factor: 28.547
Authors: Huansheng Xu; B Alexander Yi; Hao Wu; Christoph Bock; Hongcang Gu; Kathy O Lui; Joo-Hye C Park; Ying Shao; Alyssa K Riley; Ibrahim J Domian; Erding Hu; Robert Willette; John Lepore; Alexander Meissner; Zhong Wang; Kenneth R Chien Journal: Cell Res Date: 2011-11-08 Impact factor: 25.617
Authors: Luda Khait; Louise Hecker; Nicole R Blan; Garrett Coyan; Francesco Migneco; Yen-Chih Huang; Ravi K Birla Journal: J Cardiovasc Transl Res Date: 2008-01-29 Impact factor: 4.132
Authors: Ana Armiñán; Carolina Gandía; José Manuel García-Verdugo; Elisa Lledó; José Luis Mullor; José Anastasio Montero; Pilar Sepúlveda Journal: J Cardiovasc Transl Res Date: 2009-10-21 Impact factor: 4.132