Generation of ventricular-like hipsc-derived cardiomyocytes and high-quality cell preparations for calcium handling characterization

Jae Gyun Oh, Jaydev Dave, Changwon Kho, Francesca Stillitano

Research output: Contribution to journalReview articlepeer-review

Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) provide a valuable human source for studying the basic science of calcium (Ca2+) handling and signaling pathways as well as high-throughput drug screening and toxicity assays. Herein, we provide a detailed description of the methodologies used to generate high-quality iPSC-CMs that can consistently reproduce molecular and functional characteristics across different cell lines. Additionally, a method is described to reliably assess their functional characterization through the evaluation of Ca2+ handling properties. Low oxygen (O2) conditions, lactate selection, and prolonged time in culture produce high-purity and high-quality ventricular-like cardiomyocytes. Similar to isolated adult rat cardiomyocytes (ARCMs), 3-month-old iPSC-CMs exhibit higher Ca2+ amplitude, faster rate of Ca2+ reuptake (decay-tau), and a positive lusitropic response to β-adrenergic stimulation compared to day 30 iPSC-CMs. The strategy is technically simple, cost-effective, and reproducible. It provides a robust platform to model cardiac disease and for the large-scale drug screening to target Ca2+ handling proteins.

Original languageEnglish
Article numbere60135
JournalJournal of Visualized Experiments
Volume2020
Issue number155
DOIs
StatePublished - Jan 2019

Keywords

  • Calcium handling
  • Calcium transient
  • Cardiac disease modeling
  • Cardiomyocyte maturation
  • IPSCs
  • Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs)
  • Issue 155
  • Medicine

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