title: A Simple Protocol for the Myocardial Differentiation of Human iPS Cells.
abstract: We have developed a simple protocol for inducing the myocardial differentiation of human induced pluripotent stem (iPS) cells. Human iPS cell-derived embryonic bodies (EBs) were treated with a combination of activin-A, bone morphogenetic protein-4 and wnt-3a for one day in serum-free suspension culture, and were subsequently treated with noggin for three days. Thereafter, the EBs were subjected to adherent culture in media with 5% serum. All EBs were differentiated into spontaneously beating EBs, which were identified by the presence of striated muscles in transmission electron microscopy and the expression of the specific cardiomyocyte markers, NKX2-5 and TNNT2. The beating rate of the beating EBs was decreased by treatment with a rapidly activating delayed rectifier potassium current (Ikr) channel blocker, E-4031, an Ikr trafficking inhibitor, pentamidin, and a slowly activating delayed rectifier potassium current (Iks) channel blocker, chromanol 293B, and was increased by treatment with a beta-receptor agonist, isoproterenol. At a low concentration, verapamil, a calcium channel blocker, increased the beating rate of the beating EBs, while a high concentration decreased this rate. These findings suggest that the spontaneously beating EBs were myocardial cell clusters. This simple protocol for myocardial differentiation would be useful in providing a sufficient number of the beating myocardial cell clusters for studies requiring human myocardium.
authors.affiliation: Translational Research Unit, R&D Division, Kyowa Hakko Kirin Co., Ltd.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Aikawa
authors.forename: Yui
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Katsumi
authors.initials: K
authors.lastname: Takaba
mesh_terms: D028341:Activins / Q000494:pharmacology; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002352:Carrier Proteins / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology; D058732:Embryoid Bodies / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D009206:Myocardium; D010419:Pentamidine / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D026902:Potassium Channel Blockers / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D020107:Troponin T / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology; D060509:Wnt3A Protein / Q000494:pharmacology; D000099176:Noggin Protein
chemical_list: D028341:Activins; D055415:Bone Morphogenetic Protein 4; D002121:Calcium Channel Blockers; D002352:Carrier Proteins; D002839:Chromans; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; D010419:Pentamidine; D010880:Piperidines; D026902:Potassium Channel Blockers; D011725:Pyridines; D013449:Sulfonamides; D014157:Transcription Factors; D020107:Troponin T; D014700:Verapamil; D060509:Wnt3A Protein; D000099176:Noggin Protein; C525809:BMP4 protein, human; C095552:NKX2-5 protein, human; C545568:TNNT2 protein, human; C422405:activin A; C063968:E 4031; C103474:6-cyano-4-(N-ethylsulfonyl-N-methylamino)-3-hydroxy-2,2-dimethylchromane