Electrophysiological characterization of cardiomyocytes derived from human induced pluripotent stem cells.
Cardiomyocytes derived from human induced pluripotent stem cells (hiPS-CMs) hold great promise for development of in vitro research tools to assess cardiotoxicity, including QT prolongation. In the present study, we aimed to clarify the electrophysiological/pharmacological characteristics of hiPS-CMs using the patch-clamp technique. The hiPS cells were differentiated into beating cardiomyocytes by the embryoid body method. The expression of genes related to cardiac ion channels and differentiation markers in cardiomyocytes were detected by RT-PCR. Whole-cell patch-clamp recordings were performed using single hiPS-CMs dispersed from beating colonies. We confirmed voltage-dependence of major cardiac ion currents (I(Na), I(Ca), I(Kr), and I(Ks)) and pharmacological responses to ion-channel blockers. Action potential duration (APD) was prolonged by both I(Kr)/hERG and I(Ks) blockers, whereas it was shortened by an I(Ca) blocker, indicating that these ion current components contribute to action potential generation in hiPS-CMs. As for multiple ion channel blockers, terfenadine prolonged APD, but verapamil did not, results which were identical to clinically relevant pharmacological responses. These data suggest that patch-clamp assay using hiPS-CMs could be an accurate method of predicting the human cardiac responses to drug candidates. This study would be helpful in establishing an electrophysiological assay to assess the risk of drug-induced arrhythmia using hiPS-CMs.
Safety Assessment Department, Chugai Pharmaceutical Co., Ltd., 1-135 Komakado, Gotemba, Shizuoka 412-8513, Japan. hondamsk@chugai-pharm.co.jp
Masaki
M
Honda
Jumpei
J
Kiyokawa
Mitsuyasu
M
Tabo
Tomoaki
T
Inoue
D000200:Action Potentials; D002454:Cell Differentiation; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007473:Ion Channels / Q000037:antagonists & inhibitors / Q000235:genetics*; D049990:Membrane Transport Modulators / Q000494:pharmacology; D032383:Myocytes, Cardiac / Q000502:physiology*; D018408:Patch-Clamp Techniques
D007473:Ion Channels; D049990:Membrane Transport Modulators