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Myosin expression and contractile function are altered by replating stem cell-derived cardiomyocytes.

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Summary
This summary is machine-generated.

Detaching and replating human stem cell-derived cardiomyocytes (hESC-CMs and hiPSC-CMs) alters their myosin heavy chain (MyHC) expression and function. These changes in mechanosensation and gene expression are reversible and important for in vitro assays.

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Area of Science:

  • Cardiovascular Biology
  • Stem Cell Biology
  • Cellular Mechanics

Background:

  • Human ventricular cardiomyocytes (CMs) primarily express the β-myosin heavy chain (β-MyHC) isoform, crucial for contractile function.
  • Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) typically express β-MyHC after long-term culture on specific substrates.
  • The impact of mechanical manipulation on cardiomyocyte phenotype is not fully understood.

Purpose of the Study:

  • To investigate how detaching and replating hESC-CMs affects their cellular function and myosin heavy chain (MyHC) isoform expression.
  • To assess the reversibility of these changes and identify underlying molecular mechanisms.
  • To evaluate similar effects in human induced pluripotent stem cell-derived CMs (hiPSC-CMs).

Main Methods:

  • Long-term cultured hESC-CMs were enzymatically detached and replated.
  • Micromechanical assays were used to measure force-related kinetic parameters.
  • Single-cell immunofluorescence analyzed MyHC isoform expression (β-MyHC and α-MyHC).
  • RNA-sequencing and gene enrichment analysis identified affected pathways.
  • Focal adhesion kinase (FAK) role was investigated.

Main Results:

  • Replating induced significant changes in force-related kinetics and calcium transients, resembling α-MyHC myofibrils.
  • α-MyHC expression rapidly upregulated in hESC-CMs and heterogeneity increased in both hESC-CMs and hiPSC-CMs.
  • These functional and molecular changes were reversible within two weeks.
  • Gene enrichment analysis highlighted alterations in mechanosensation/transduction pathways, including integrin signaling and FAK.

Conclusions:

  • Detachment and replating substantially alter gene expression, MyHC composition, and function of stem cell-derived CMs.
  • Mechanosensation and mechanotransduction pathways are significantly affected by these procedures.
  • These induced changes must be considered for accurate interpretation of downstream in vitro assays using stem cell-derived CMs.