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

Joe Z Zhang1, Vittavat Termglinchan1, Ning-Yi Shao1

  • 1Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA; Division of Cardiology, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.

Cell Stem Cell
|March 19, 2019
PubMed

Insights

Researchers created a human induced pluripotent stem cell (hiPSC) reporter line to distinguish cardiac cell types. This tool aids in isolating specific cardiac subpopulations for drug testing and disease modeling.

Area of Science:

  • Cardiovascular Biology
  • Stem Cell Research
  • Developmental Biology

Background:

  • Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) exhibit significant heterogeneity due to diverse cardiac lineages.
  • Understanding and isolating these distinct cardiac lineages is crucial for advancing cardiovascular research and regenerative medicine.

Purpose of the Study:

  • To develop a novel hiPSC reporter line for delineating cardiac lineages and isolating specific subpopulations.
  • To characterize the identified subpopulations and validate their utility in drug testing and disease modeling.

Main Methods:

  • Generation of a hiPSC TBX5Clover2 and NKX2-5TagRFP double reporter line.
  • Isolation and molecular analysis of four subpopulations based on differential TBX5 and NKX2-5 expression.
  • Genetic and functional characterization of isolated subpopulations, including identification of CORIN as a cell-surface marker.

Main Results:

  • Four distinct subpopulations (TBX5+NKX2-5+, TBX5+NKX2-5-, TBX5-NKX2-5+, TBX5-NKX2-5-) were identified, mimicking cardiac lineages.
  • Each subpopulation demonstrated differentiation into specific cardiac cell types.
  • CORIN was identified as a cell-surface marker for the TBX5+NKX2-5+ subpopulation, enabling its isolation.

Conclusions:

  • The developed hiPSC reporter line effectively delineates cardiac lineages and allows for the isolation of specific subpopulations.
  • Lineage-specific CMs facilitate precise drug testing, offering a valuable tool for cardiac research, tissue engineering, and disease modeling.

Keywords:
CORINNKX2-5TBX5cardiomyocyte subtypesendothelial cell lineageepicardial lineagehiPSC double reporterhuman first and second heart fieldprecise drug testingpurification

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