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Related Experiment Video

Updated: Aug 5, 2025

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts
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Optimized protocol for direct cardiac reprogramming in mice using Ascl1 and Mef2c.

Haofei Wang1, Benjamin Keepers1, Jiandong Liu1

  • 1The McAllister Heart Institute, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Pathology and Laboratory Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

STAR Protocols
|March 29, 2023
PubMed
Summary
This summary is machine-generated.

This study introduces a new protocol for direct cardiac reprogramming, converting fibroblasts into cardiomyocyte-like cells (iCMs) using Ascl1 and Mef2c. The method efficiently generates iCMs with key cardiac characteristics in mice.

Keywords:
Cell BiologyCell CultureCell IsolationDevelopmental BiologyModel Organisms

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Area of Science:

  • Cardiovascular Research
  • Stem Cell Biology
  • Molecular Cardiology

Background:

  • Direct cardiac reprogramming offers a potential strategy for cardiac repair by generating cardiomyocyte-like cells (iCMs) without intermediate progenitor stages.
  • Fibroblast-to-cardiomyocyte conversion is a key focus in regenerative medicine for treating heart disease.

Purpose of the Study:

  • To present a detailed protocol for direct cardiac reprogramming of fibroblasts into iCMs in mice.
  • To demonstrate the efficiency and characteristics of iCMs generated using the Ascl1 and Mef2c factors.

Main Methods:

  • Isolation of primary neonatal mouse cardiac fibroblasts.
  • Preparation of retroviruses encoding the reprogramming factors Ascl1 and Mef2c.
  • Execution of direct cardiac reprogramming protocol in vitro.

Main Results:

  • Successful generation of cardiomyocyte-like cells (iCMs) from cardiac fibroblasts.
  • Characterization of iCMs showing cardiomyocyte-like sarcomere structure.
  • Confirmation of iCMs exhibiting cardiomyocyte-specific gene expression and functional calcium flux.

Conclusions:

  • The presented protocol enables efficient direct cardiac reprogramming using Ascl1 and Mef2c.
  • The generated iCMs possess key functional and structural properties of native cardiomyocytes.
  • This method provides a valuable tool for studying cardiac reprogramming and potential therapeutic applications.