A Human Induced Pluripotent Stem Cell−Derived Cardiomyocyte (hiPSC‐CM) Multielectrode Array Assay for Preclinical Cardiac Electrophysiology Safety Screening

Kate Harris1

1 Safety Pharmacology, GlaxoSmithKline, David Jack Centre for R&D, Ware, Hertfordshire, United Kingdom
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 11.18
DOI:  10.1002/0471141755.ph1118s71
Online Posting Date:  December, 2015
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Abstract

Cardiotoxicity is a leading cause of compound attrition during drug development. Preclinical models used to assess the risk for compound‐induced effects on cardiac electrophysiology largely rely on animals that can differ in terms of sensitivity and specificity to the targeted clinical response. There is currently no in vitro human cardiomyocyte model for routine preclinical compound screening, as adult human cardiac tissue is unsuitable for such screening. The commercial availability of human induced pluripotent stem cell−derived cardiomyocytes (hiPSC‐CMs) makes possible the development of assays for assessing compound‐induced effects on cardiac function in a human cardiomyocyte‐like model. Using multielectrode array (MEA) technology with hiPSC‐CMs provides a facile screen for compound‐induced effects on cardiac electrophysiology. The MEA data generated from hiPSC‐CMs correlate well with the results of conventional preclinical assays and clinical findings. Described in this unit is a technique for measuring extracellular field potentials from hiPSC‐CMs using MEA technology to screen for compound‐induced effects on cardiac electrophysiology. © 2015 by John Wiley & Sons, Inc.

Keywords: human induced pluripotent stem cell−derived cardiomyocytes; multielectrode array; preclinical; cardiac electrophysiology; cardiotoxicity; arrhythmia

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Measurement of Field Potentials from hiPSC‐CM Monolayers Using Mea Technology
  • Support Protocol 1: Preparation of iCell Cardiomyocyte Monolayers on Mea Plates
  • Support Protocol 2: Analysis of MEA Recording Data
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Measurement of Field Potentials from hiPSC‐CM Monolayers Using Mea Technology

  Materials
  • iCell cardiomyocyte monolayers on MEA plates (see protocol 2)
  • Serum‐free medium (see recipe)
  • Test compound(s) of interest
  • Control compounds:
    • Captopril (Sigma‐Aldrich, cat. no. C4042)
    • E4031 (Enzo Life Sciences, cat. no. BML‐KC158‐0025)
    • Nifedipine (Sigma‐Aldrich, cat. no. N7634)
  • Dimethyl sulfoxide (DMSO)
  • MEA2100‐System (Multi Channel Systems)
  • Computer running MC_Rack software (Multi Channel Systems)

Support Protocol 1: Preparation of iCell Cardiomyocyte Monolayers on Mea Plates

  Materials
  • Sterile water (e.g., Sigma‐Aldrich, cat. no. W3500)
  • EmbryoMax 0.1% (w/v) gelatin solution (Millipore, cat. no. ES‐006‐B)
  • iCell Cardiomyocytes Plating Medium (Cellular Dynamics International), room temperature
  • iCell Cardiomyocytes (Cellular Dynamics International, cat. no. CMC‐100‐010‐001, >4.0 × 106 viable cells), stored in liquid nitrogen
  • Trypan blue (Gibco, cat. no. 15250)
  • iCell Cardiomyocytes Maintenance Medium (Cellular Dynamics International), 37°C
  • 50 mg/ml gentamicin (Gibco, cat. no. 15750)
  • Single‐well EcoMEA plates (Multi Channel Systems, cat. no. 60EcoMEA‐Glass‐gr)
  • 100‐mm sterile Petri dishes (e.g., Grenier Bio‐One, cat. no. 632161)
  • Laboratory tissues (e.g., Kimwipes)
  • Hemocytometer or automated cell counter
  • 50‐ml sterile centrifuge tube (e.g., Corning Life Sciences)
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Figures

Videos

Literature Cited

Literature Cited
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