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Advanced Single-Cell Mapping Reveals that in hESC Cardiomyocytes Contraction Kinetics and Action Potential Are

Natalie Weber1, Kathrin Kowalski1, Tim Holler1

  • 1Institute of Molecular and Cell Physiology, Hannover Medical School, Carl-Neuberg Straße 1, 30625 Hannover, Germany.

Stem Cell Reports
|April 18, 2020
PubMed

Insights

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) show independent regulation of genes. Myosin heavy chain (MyHC) expression does not influence their contraction kinetics or electrical activity.

Area of Science:

  • Cardiology
  • Stem Cell Biology
  • Molecular Biology

Background:

  • Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are valuable models for studying cardiomyocyte (CM) function and disease.
  • Ventricular CMs are characterized by slow β-myosin heavy chain (MyHC) expression, distinct from atrial CMs' fast α-MyHC.
  • Understanding the relationship between MyHC isoform expression and CM function is crucial.

Purpose of the Study:

  • To investigate the influence of α-MyHC versus β-MyHC expression on the contractile and electrical properties of hPSC-CMs at the single-cell level.
  • To determine if MyHC isoform ratios correlate with specific functional characteristics in individual hPSC-CMs.

Main Methods:

  • Development of a novel single-cell mapping technique to simultaneously assess functional parameters and MyHC isoform expression (protein and mRNA) in the same hPSC-CMs.
  • Multiparametric, cell-by-cell analysis of contractile kinetics and electrical activity.
  • Quantitative analysis of α-MyHC and β-MyHC expression at both mRNA and protein levels.

Main Results:

  • Surprisingly, the relative expression of α-MyHC versus β-MyHC was not associated with specific contractile or electrophysiological properties within individual hPSC-CMs.
  • The study revealed that genes related to electrical activity, contraction, calcium handling, and MyHC expression are independently regulated in hPSC-CMs.
  • No direct correlation was found between MyHC isoform ratios and functional phenotypes at the single-cell level.

Conclusions:

  • The relative expression of α-MyHC and β-MyHC does not dictate the contractile or electrophysiological characteristics of individual hPSC-CMs.
  • Gene expression related to key cardiomyocyte functions, including MyHC isoforms, appears to be independently regulated in hPSC-CMs.
  • These findings suggest a complex regulatory network governing hPSC-CM function, separate from simple MyHC isoform ratios.

Keywords:
MYH6MYH7action potentialcardiac myosin heavy chainhuman embryonic stem cell-derived cardiomyocytesmaturationsingle-cell mapping techniquetwitch contractions

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