In vitro generation of heart field-specific cardiac progenitor cells

Research output: Contribution to journalArticle

Abstract

Pluripotent stem cells offer great potential for understanding heart development and disease and for regenerative medicine. While recent advances in developmental cardiology have led to generating cardiac cells from pluripotent stem cells, it is unclear if the two cardiac fields-the first and second heart fields (FHF and SHF) — are induced in pluripotent stem cells systems. To address this, we generated a protocol for in vitro specification and isolation of heart field-specific cardiac progenitor cells. We used embryonic stem cells lines carrying Hcn4-GFP and Tbx1-Cre; Rosa-RFP reporters of the FHF and the SHF, respectively, and live cell immunostaining of the cell membrane protein Cxcr4, a SHF marker. With this approach, we generated progenitor cells which recapitulate the functional properties and transcriptome of their in vivo counterparts. Our protocol can be utilized to study early specification and segregation of the two heart fields and to generate chamber-specific cardiac cells for heart disease modelling. Since this is an in vitro organoid system, it may not provide precise anatomical information. However, this system overcomes the poor accessibility of gastrulation-stage embryos and can be upscaled for high-throughput screens.

Original languageEnglish (US)
Article numbere59826
JournalJournal of Visualized Experiments
Volume2019
Issue number149
DOIs
StatePublished - Jul 1 2019

Fingerprint

Pluripotent Stem Cells
Stem cells
Stem Cells
Heart Diseases
Organoids
Induced Pluripotent Stem Cells
Gastrulation
Regenerative Medicine
Embryonic Stem Cells
Cardiology
Transcriptome
Membrane Proteins
Specifications
Embryonic Structures
Cell Line
Cell membranes
Throughput
In Vitro Techniques
Proteins
caN protocol

Keywords

  • Cardiac progenitors
  • Cardiac spheroids
  • Cxcr4
  • Developmental Biology
  • Hcn4
  • Heart fields
  • Issue 149
  • Stem cells
  • Tbx1

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

In vitro generation of heart field-specific cardiac progenitor cells. / Tampakakis, Emmanouil; Miyamoto, Matthew; Kwon, Chulan.

In: Journal of Visualized Experiments, Vol. 2019, No. 149, e59826, 01.07.2019.

Research output: Contribution to journalArticle

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