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Updated: Sep 24, 2025
In Vitro Generation of Heart Field-specific Cardiac Progenitor Cells
Published on: July 3, 2019
In Vitro Generation of Heart Field Specific Cardiomyocytes
Arash Pezhouman1,2, Ngoc B Nguyen1,2,3, Allison Shevtsov1
1Division of Cardiology, Department of Internal Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
Insights
This study efficiently generates first heart field (FHF) and second heart field (SHF) cardiomyocytes from human embryonic stem cells. This method aids in developing cell therapies for heart damage, particularly after myocardial infarction (MI).
Area of Science:
- Cardiovascular Biology
- Stem Cell Biology
- Developmental Biology
Background:
- Myocardial infarction (MI) causes irreversible loss of cardiomyocytes (CMs), especially in the left ventricle (LV).
- Left ventricle CMs originate from first heart field (FHF) progenitors, while right ventricle CMs derive from the second heart field (SHF).
- Human embryonic stem cells (hESCs) are crucial for studying cardiac development and generating cell therapy candidates.
Purpose of the Study:
- To develop an efficient method for generating FHF and SHF CMs from hESCs.
- To utilize a double reporter hESC line (TBX5/NKX2-5) for CM lineage tracking.
- To provide a platform for comparative analyses and in vitro studies for cardiovascular disease research.
Main Methods:
- Generation of FHF and SHF CMs using a TBX5/NKX2-5 double reporter hESC line.
- Step-by-step protocol for efficient CM subtype generation.
- Application of the method to non-genetically modified hESC lines for enrichment.
Main Results:
- Efficient generation of distinct FHF and SHF cardiomyocyte populations.
- Demonstration of a protocol applicable to various hESC lines.
- Establishment of a method to enrich specific CM subtypes.
Conclusions:
- The described method enables efficient isolation of FHF and SHF CMs.
- This approach supports research into cardiac lineage commitment and cell therapy development.
- Enriched CM subtypes facilitate understanding of heart development and treatment strategies for MI.
Abstract:
Myocardial infarction (MI) can lead to irreversible loss of cardiomyocytes (CMs), primarily localized to the left ventricle (LV) of the heart. The CMs of the LV are predominantly derived from first heart field (FHF) progenitors, whereas the majority of CMs within the right ventricle originate from the second heart field (SHF) during early cardiogenesis. Human embryonic stem cells (hESCs) serve as a valuable source of CMs for understanding early cardiac development and lineage commitment of CMs within these two heart fields that ultimately enable the development of more effective candidates for cell therapy. An ideal candidate may be FHF CMs that share the same ontogeny with the LV CMs that die after MI. We previously generated a double reporter hESC line that utilizes two important cardiac transcription factors, TBX5 and NKX2-5. TBX5 marks FHF progenitors and CMs, while NKX2-5 is expressed in nearly all myocytes of the developing heart. Here, we describe a step-by-step approach to efficiently generate FHF and SHF CMs using this double reporter hESC line. In addition, this approach can be applied to any non-genetically modified hESC lines to enrich FHF and SHF CMs. Obtaining enriched populations of these two CM subtypes provides a platform for downstream comparative analyses and in vitro studies to facilitate a deeper understanding of cardiovascular lineage commitment and the development of more effective candidates for cell therapy to treat diseases or defects that affect specific regions of the heart.
