Comparative analysis of media effects on human induced pluripotent stem cell-derived cardiomyocytes in proarrhythmia risk assessment

Derek Schocken; Jayna Stohlman; Jose Vicente; Dulciana Chan; Dakshesh Patel; Murali Krishna Matta; Vikram Patel; Mathew Brock; Daniel Millard; James Ross; David G Strauss; Ksenia Blinova

doi:10.1016/j.vascn.2017.11.002

Comparative analysis of media effects on human induced pluripotent stem cell-derived cardiomyocytes in proarrhythmia risk assessment

J Pharmacol Toxicol Methods. 2018 Mar-Apr:90:39-47. doi: 10.1016/j.vascn.2017.11.002. Epub 2017 Nov 16.

Authors

Affiliations

¹ US Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Biomedical Physics, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Derek.schocken@fda.hhs.gov.
² US Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Biomedical Physics, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Jayna.stohlman@fda.hhs.gov.
³ US Food and Drug Administration, Center for Drug Evaluation and Research, Office of New Drugs, Division of Cardiovascular and Renal Products, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Jose.vicenteruiz@fda.hhs.gov.
⁴ US Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Biomedical Physics, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Dulciana.chan@fda.hhs.gov.
⁵ US Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Biomedical Physics, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Dakshsesh.patel1@fda.hhs.gov.
⁶ US Food and Drug Administration, Center for Drug Evaluation and Research, Office of Clinical Pharmacology, Division of Applied Regulatory Science, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Murali.matta@fda.hhs.gov.
⁷ US Food and Drug Administration, Center for Drug Evaluation and Research, Office of Clinical Pharmacology, Division of Applied Regulatory Science, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Vikram.Patel@fda.hhs.gov.
⁸ Axion BioSystems, Inc., 1819 Peachtree Road NE, Suite 350, Atlanta, GA 30309, USA. Electronic address: Brockm3@gene.com.
⁹ Axion BioSystems, Inc., 1819 Peachtree Road NE, Suite 350, Atlanta, GA 30309, USA. Electronic address: dmillard@axion-biosystems.com.
¹⁰ Axion BioSystems, Inc., 1819 Peachtree Road NE, Suite 350, Atlanta, GA 30309, USA. Electronic address: jross@axionbio.com.
¹¹ US Food and Drug Administration, Center for Drug Evaluation and Research, Office of Clinical Pharmacology, Division of Applied Regulatory Science, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: David.strauss@fda.hhs.gov.
¹² US Food and Drug Administration, Center for Devices and Radiological Health, Office of Science and Engineering Laboratories, Division of Biomedical Physics, 10903 New Hampshire Ave, Silver Spring, MD 20993, USA. Electronic address: Ksenia.blinova@fda.hhs.gov.

PMID: 29155283
DOI: 10.1016/j.vascn.2017.11.002

Abstract

Introduction: Cardiotoxicity assessment using human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) forms a key component of the Comprehensive in Vitro Proarrhythmia Assay (CiPA). A potentially impactful factor on iPSC-CM testing is the presence of serum in the experimental media. Generally, serum-free media is used to most accurately reproduce "free" drug concentration. However, caution is needed; drug solubility and cardiomyocyte electrophysiology could be affected by media formulation, potentially impacting interpretation of drug-induced effects.

Methods: Effects of 25 drugs on properties of spontaneous field potentials in iPSC-CMs were assayed using a high-throughput microelectrode array (MEA) in two media formulations: serum-containing and serum-free. Comparative analysis was conducted on rate-corrected field potential duration (FPDc) and prevalence of arrhythmic events. Further MEA experiments were conducted, varying percentages of serum as well as carbon substrate components. Comparative LC-MS/MS analysis was done on two compounds to evaluate drug concentrations.

Results: In serum-free media, 9 drugs prolonged FPDc. In serum-containing, 11 drugs prolonged FPDc. Eighteen drugs induced arrhythmias, 8 of these induced arrhythmias at lower concentrations in serum-containing media. At the highest non-arrhythmic concentrations, 13 of 25 drugs exhibited significant differences in FPDc prolongation/shortening between the media. Increasing fractions of serum in media yielded higher FPDc measurements. LC-MS/MS analysis of moxifloxacin and quinidine showed higher concentrations in serum-containing media.

Discussion: The present study highlights media formulation as an important consideration for cardiac safety testing with iPSC-CMs. Results described here suggest that media formulation influences both compound availability and baseline electrophysiological properties. Special attention should be paid to media for future iPSC-CM assays.

Published by Elsevier Inc.

Publication types

Comparative Study

MeSH terms

Arrhythmias, Cardiac / chemically induced*
Arrhythmias, Cardiac / metabolism
Cardiotoxicity / etiology*
Cardiotoxicity / metabolism
Cells, Cultured
Culture Media / adverse effects*
Culture Media / pharmacology*
Electrophysiological Phenomena / drug effects
Humans
Induced Pluripotent Stem Cells / drug effects*
Long QT Syndrome / chemically induced
Long QT Syndrome / metabolism
Myocytes, Cardiac / drug effects*
Myocytes, Cardiac / metabolism
Risk Assessment
Serum / metabolism*

Substances

Culture Media