title: Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals.
abstract: Although myocyte cell transplantation studies have suggested a promising therapeutic potential for myocardial infarction, a major obstacle to the development of clinical therapies for myocardial repair is the difficulties associated with obtaining relatively homogeneous ventricular myocytes for transplantation. Human embryonic stem cells (hESCs) are a promising source of cardiomyocytes. Here we report that retinoid signaling regulates the fate specification of atrial versus ventricular myocytes during cardiac differentiation of hESCs. We found that both Noggin and the pan-retinoic acid receptor antagonist BMS-189453 (RAi) significantly increased the cardiac differentiation efficiency of hESCs. To investigate retinoid functions, we compared Noggin+RAi-treated cultures with Noggin+RA-treated cultures. Our results showed that the expression levels of the ventricular-specific gene IRX-4 were radically elevated in Noggin+RAi-treated cultures. MLC-2V, another ventricular-specific marker, was expressed in the majority of the cardiomyocytes in Noggin+RAi-treated cultures, but not in the cardiomyocytes of Noggin+RA-treated cultures. Flow cytometry analysis and electrophysiological studies indicated that with 64.7 ± 0.88% (mean ±s.e.m) cardiac differentiation efficiency, 83% of the cardiomyocytes in Noggin+RAi-treated cultures had embryonic ventricular-like action potentials (APs). With 50.7 ± 1.76% cardiac differentiation efficiency, 94% of the cardiomyocytes in Noggin+RA-treated cultures had embryonic atrial-like APs. These results were further confirmed by imaging studies that assessed the patterns and properties of the Ca(2+) sparks of the cardiomyocytes from the two cultures. These findings demonstrate that retinoid signaling specifies the atrial versus ventricular differentiation of hESCs. This study also shows that relatively homogeneous embryonic atrial- and ventricular-like myocyte populations can be efficiently derived from hESCs by specifically regulating Noggin and retinoid signals.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, China.
authors.forename: Qiangzhe
authors.initials: Q
authors.lastname: Zhang
authors.forename: Junjie
authors.initials: J
authors.lastname: Jiang
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Han
authors.forename: Qi
authors.initials: Q
authors.lastname: Yuan
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Zhang
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Xu
authors.forename: Henghua
authors.initials: H
authors.lastname: Cao
authors.forename: Qingzhang
authors.initials: Q
authors.lastname: Meng
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.forename: Tian
authors.initials: T
authors.lastname: Tian
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Pu
authors.initials: P
authors.lastname: Li
authors.forename: Jurgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Guangju
authors.initials: G
authors.lastname: Ji
authors.forename: Yue
authors.initials: Y
authors.lastname: Ma
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D001483:Base Sequence; D015153:Blotting, Western; D024722:Cardiac Myosins / Q000096:biosynthesis / Q000235:genetics; D002352:Carrier Proteins / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006325:Heart Atria / Q000196:embryology / Q000378:metabolism*; D006352:Heart Ventricles / Q000196:embryology / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism*; D018994:Myosin Light Chains / Q000096:biosynthesis / Q000235:genetics; D016133:Polymerase Chain Reaction; D012176:Retinoids / Q000378:metabolism* / Q000494:pharmacology; D015398:Signal Transduction; D000099176:Noggin Protein
chemical_list: D024722:Cardiac Myosins; D002352:Carrier Proteins; D018398:Homeodomain Proteins; D018994:Myosin Light Chains; D012176:Retinoids; D000099176:Noggin Protein; C423184:BMS 189453; C485533:IRX4 protein, human; C107184:myosin light chain 2