title: Improvement of acquisition and analysis methods in multi-electrode array experiments with iPS cell-derived cardiomyocytes.
abstract: BACKGROUND
  Multi-electrode array (MEA) systems and human induced pluripotent stem (iPS) cell-derived cardiomyocytes are frequently used to characterize the electrophysiological effects of drug candidates for the prediction of QT prolongation and proarrhythmic potential. However, the optimal experimental conditions for obtaining reliable experimental data, such as high-pass filter (HPF) frequency and cell plating density, remain to be determined.
  
  
  METHODS
  Extracellular field potentials (FPs) were recorded from iPS cell-derived cardiomyocyte sheets by using the MED64 and MEA2100 multi-electrode array systems. Effects of HPF frequency (0.1 or 1Hz) on FP duration (FPD) were assessed in the presence and absence of moxifloxacin, terfenadine, and aspirin. The influence of cell density on FP characteristics recorded through a 0.1-Hz HPF was examined. The relationship between FP and action potential (AP) was elucidated by simultaneous recording of FP and AP using a membrane potential dye.
  
  
  RESULTS
  Many of the FP waveforms recorded through a 1-Hz HPF were markedly deformed and appeared differentiated compared with those recorded through a 0.1-Hz HPF. The concentration-response curves for FPD in the presence of terfenadine reached a steady state at concentrations of 0.1 and 0.3μM when a 0.1-Hz HPF was used. In contrast, FPD decreased at a concentration of 0.3μM with a characteristic bell-shaped concentration-response curve when a 1-Hz HPF was used. The amplitude of the first and second peaks in the FP waveform increased with increasing cell plating density. The second peak of the FP waveform roughly coincided with AP signal at 50% repolarization, and the negative deflection at the second peak of the FP waveform in the presence of E-4031 corresponded to early afterdepolarization and triggered activity.
  
  
  CONCLUSIONS
  FP can be used to assess the QT prolongation and proarrhythmic potential of drug candidates; however, experimental conditions such as HPF frequency are important for obtaining reliable data.
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Non-Clinical Evaluation Expert Committee, Drug Evaluation Committee, Japan Pharmaceutical Manufacturers Association (JPMA), 2-3-11 Nihonbashi-Honcho, Chuo-ku, Tokyo 103-0023, Japan; Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Nippon Shinyaku Co., Ltd., 14, Nishinosho-Monguchi-cho, Kisshoin, Minami-ku, Kyoto 601-8550, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Keiichi
authors.initials: K
authors.lastname: Asakura
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Nippon Shinyaku Co., Ltd., 14, Nishinosho-Monguchi-cho, Kisshoin, Minami-ku, Kyoto 601-8550, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Seiji
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan. Electronic address: http://jicsa.org/
authors.forename: Atsuko
authors.initials: A
authors.lastname: Ojima
authors.affiliation: Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Non-Clinical Evaluation Expert Committee, Drug Evaluation Committee, Japan Pharmaceutical Manufacturers Association (JPMA), 2-3-11 Nihonbashi-Honcho, Chuo-ku, Tokyo 103-0023, Japan; Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan. Electronic address: t2-taniguchi@hhc.eisai.co.jp.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Taniguchi
authors.affiliation: Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Non-Clinical Evaluation Expert Committee, Drug Evaluation Committee, Japan Pharmaceutical Manufacturers Association (JPMA), 2-3-11 Nihonbashi-Honcho, Chuo-ku, Tokyo 103-0023, Japan; Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan. Electronic address: http://jicsa.org/
authors.forename: Norimasa
authors.initials: N
authors.lastname: Miyamoto
authors.affiliation: Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Taisho Pharmaceutical Co., Ltd., 1-403 Yoshino-cho, Kita-ku, Saitama, Saitama 331-9530, Japan. Electronic address: http://csahi.org/en/
authors.forename: Chiaki
authors.initials: C
authors.lastname: Nakamori
authors.affiliation: Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Taisho Pharmaceutical Co., Ltd., 1-403 Yoshino-cho, Kita-ku, Saitama, Saitama 331-9530, Japan. Electronic address: http://csahi.org/en/
authors.forename: Chiho
authors.initials: C
authors.lastname: Nagasawa
authors.affiliation: Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; LSI Medience Corporation, 13-4 Uchikanda 1-chome, Chiyoda-ku, Tokyo 101-8517, Japan. Electronic address: http://csahi.org/en/
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Kitamura
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; LSI Medience Corporation, 13-4 Uchikanda 1-chome, Chiyoda-ku, Tokyo 101-8517, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Osada
authors.affiliation: Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Sumitomo Dainippon Pharma Co., Ltd., 3-1-98 Kasugade-naka, Konohana-Ku, Osaka 554-0022, Japan. Electronic address: http://csahi.org/en/
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Honda
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Astellas Pharma Inc., 2-1-6 Kashima, Yodogawa-ku, Osaka 532-8514, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Chieko
authors.initials: C
authors.lastname: Kasai
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Consortium for Safety Assessment using Human iPS Cells (CSAHi), Japan; Ono Pharmaceutical Co., Ltd., 50-10 Yamagishi Mikuni-cho, Sakaishi, Fukui 913-8538, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ando
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; National Institute of Health Sciences (NIHS), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Yasunari
authors.initials: Y
authors.lastname: Kanda
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; National Institute of Health Sciences (NIHS), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Yuko
authors.initials: Y
authors.lastname: Sekino
authors.affiliation: Japanese Safety Pharmacology Society (JSPS), 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan; Japan iPS Cardiac Safety Assessment (JiCSA), 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; Eisai Co., Ltd., 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan. Electronic address: http://www.j-sps.org/
authors.forename: Kohei
authors.initials: K
authors.lastname: Sawada
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D001145:Arrhythmias, Cardiac / Q000139:chemically induced* / Q000175:diagnosis; D001241:Aspirin / Q000008:administration & dosage / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D024841:Fluoroquinolones / Q000008:administration & dosage / Q000633:toxicity; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008133:Long QT Syndrome / Q000139:chemically induced* / Q000175:diagnosis; D000077266:Moxifloxacin; D032383:Myocytes, Cardiac / Q000187:drug effects*; D010880:Piperidines / Q000008:administration & dosage / Q000633:toxicity; D011725:Pyridines / Q000008:administration & dosage / Q000633:toxicity; D016593:Terfenadine / Q000008:administration & dosage / Q000633:toxicity
chemical_list: D024841:Fluoroquinolones; D010880:Piperidines; D011725:Pyridines; C063968:E 4031; D016593:Terfenadine; D001241:Aspirin; D000077266:Moxifloxacin
keywords: Action potential; Field potential; High-pass filter; Human induced pluripotent stem cell-derived cardiomyocytes; Membrane potential dye; Multi-electrode array