Influence of Electromechanical Activity on Cardiac Differentiation of Mouse Embryonic Stem Cells

Worawan Limpitikul, Nicolas Christoforou, Susan A. Thompson, John D. Gearhart, Leslie Tung, Elizabeth A. Lipke

Research output: Contribution to journalArticle

Abstract

opment. Electrical pacing was applied to confluent monolayers of mESC-CMs during late-stage differentiation (days 16-18). Alternatively, spontaneous contraction was suppressed by (a) blocking ion currents with CsCl (HCN channel), trazodone (T-type Ca2+ channel), or both CsCl and trazodone on days 11-18; or (b) applying blebbistatin (excitation-contraction uncoupler) on days 11-14. Electrophysiological properties and gene expression were examined on day 19 and 18, respectively. Optical mapping revealed no significant difference in conduction velocity (CV) in paced vs. non-paced monolayers, nor were there significant changes in gene expression of connexin-43, Na-Ca exchanger (NCX), or myosin heavy chain (MHC). However, CV variability among differentiation batches and CV heterogeneity within individual monolayers were significantly lower in paced mESC-CMs. Alternatively, while the four drug treatments suppressed contraction with varying degrees (up to complete inhibition), there was no significant difference in CV for any of the treatments compared with controls. Trazodone treatment significantly reduced CV variability as compared to controls, whereas CsCl treatment significantly reduced CV heterogeneity. Distinct changes in gene expression of connexin-43, MHC, HCNl, Cav3. 1/3.2 were not observed. Electrical pacing, but not suppression of spontaneous contraction, during late-stage differentiation reduces the intrinsic variability of CV among differentiation batches and across individual monolayers, which can be beneficial in the application of ESCs for myocardial tissue repair.

Original languageEnglish (US)
Pages (from-to)179-193
Number of pages15
JournalCardiovascular Engineering and Technology
Volume1
Issue number3
DOIs
Publication statusPublished - Sep 2010

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Keywords

  • Cardiac regeneration
  • Cell culture
  • Electrical stimulation
  • Electrophysiology
  • Optical mapping

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cardiology and Cardiovascular Medicine

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