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In Vitro Generation of Heart Field-specific Cardiac Progenitor Cells
09:29

In Vitro Generation of Heart Field-specific Cardiac Progenitor Cells

Published on: July 3, 2019

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In Vitro Generation of Heart Field-specific Cardiac Progenitor Cells

Emmanouil Tampakakis1, Matthew Miyamoto1, Chulan Kwon2

  • 1Division of Cardiology, Department Medicine, Johns Hopkins School of Medicine.

Journal of Visualized Experiments : Jove
|July 23, 2019
PubMed

Insights

Researchers developed a new protocol to isolate cardiac progenitor cells from pluripotent stem cells, distinguishing between the first and second heart fields. This method aids in studying heart development and generating cells for disease modeling.

Area of Science:

  • Cardiology
  • Developmental Biology
  • Stem Cell Biology

Background:

  • Pluripotent stem cells (PSCs) are crucial for understanding heart development and disease.
  • Generating specific cardiac cell types from PSCs is vital for regenerative medicine.
  • Distinguishing between the first and second heart fields (FHF and SHF) in PSC systems remains a challenge.

Purpose of the Study:

  • To develop a protocol for in vitro specification and isolation of FHF and SHF cardiac progenitor cells from PSCs.
  • To validate if PSC-derived progenitor cells mimic in vivo FHF and SHF characteristics.
  • To enable studies on early heart field segregation and disease modeling.

Main Methods:

  • Utilized embryonic stem cell lines with specific reporter genes (Hcn4-GFP for FHF, Tbx1-Cre; Rosa-RFP for SHF).
  • Employed live cell immunostaining for Cxcr4, a Second Heart Field marker.
  • Characterized generated progenitor cells for functional properties and transcriptome.

Main Results:

  • Successfully generated cardiac progenitor cells from PSCs that recapitulate FHF and SHF properties.
  • The protocol allows for the isolation of distinct FHF and SHF progenitor populations.
  • Generated cells exhibit functional properties and transcriptomes similar to their in vivo counterparts.

Conclusions:

  • The developed protocol effectively specifies and isolates FHF and SHF cardiac progenitor cells from PSCs.
  • This system provides a valuable tool for studying early heart field development and segregation.
  • The protocol facilitates the generation of chamber-specific cardiac cells for disease modeling and high-throughput screening.

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