A Human iPSC Double-Reporter System Enables Purification of Cardiac Lineage Subpopulations with Distinct Function and Drug Response Profiles

Cell Stem Cell. 2019 May 2;24(5):802-811.e5. doi: 10.1016/j.stem.2019.02.015. Epub 2019 Mar 14.


The diversity of cardiac lineages contributes to the heterogeneity of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs). Here, we report the generation of a hiPSC TBX5Clover2 and NKX2-5TagRFP double reporter to delineate cardiac lineages and isolate lineage-specific subpopulations. Molecular analyses reveal that four different subpopulations can be isolated based on the differential expression of TBX5 and NKX2-5, TBX5+NKX2-5+, TBX5+NKX2-5-, TBX5-NKX2-5+, and TBX5-NKX2-5-, mimicking the first heart field, epicardial, second heart field, and endothelial lineages, respectively. Genetic and functional characterization indicates that each subpopulation differentiates into specific cardiac cells. We further identify CORIN as a cell-surface marker for isolating the TBX5+NKX2-5+ subpopulation and demonstrate the use of lineage-specific CMs for precise drug testing. We anticipate that this tool will facilitate the investigation of cardiac lineage specification and isolation of specific cardiac subpopulations for drug screening, tissue engineering, and disease modeling.

Keywords: CORIN; NKX2-5; TBX5; cardiomyocyte subtypes; endothelial cell lineage; epicardial lineage; hiPSC double reporter; human first and second heart field; precise drug testing; purification.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism*
  • Biomarkers, Pharmacological
  • Cell Differentiation
  • Cell Lineage
  • Cell Separation / methods*
  • Cells, Cultured
  • Genes, Reporter
  • Homeobox Protein Nkx-2.5 / genetics
  • Homeobox Protein Nkx-2.5 / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / physiology*
  • Myocardium / cytology*
  • Myocytes, Cardiac / physiology*
  • Serine Endopeptidases / metabolism*
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism
  • Tissue Engineering


  • Biomarkers
  • Biomarkers, Pharmacological
  • Homeobox Protein Nkx-2.5
  • NKX2-5 protein, human
  • T-Box Domain Proteins
  • T-box transcription factor 5
  • CORIN protein, human
  • Serine Endopeptidases