How do you characterize iCell Cardiac Progenitor Cells?

iCell® Cardiac Progenitor Cells are representative of the earliest stages of mesodermal commitment to the cardiac lineage. These cells are capable of both proliferation and differentiation, (primarily to cardiomyocytes, but also to vascular smooth muscle and endothelial cells), and provide a model for investigations into both cardiac development and regeneration, capable of spontaneous beating after differentiation into cardiomyocytes.

After two days in culture there can be morphology changes, indicative of spontaneous differentiation, but the cultures will be mostly cTNT– / SMA+ based on flow cytometry. Notably the proliferation assay in the Application Protocol was developed to end two days after plating. At that time point cells still maintain the KDR+ / cKIT– profile and expression NKX2.5.

You can use immunocytochemistry techniques to measure cellular markers as confirmation of successful direction of the cell growth.

Abbreviation Description Localization
KDR
(VEGFR2)
kinase insert domain receptor (vascular endothelial growth factor receptor 2) Cardiac mesoderm
NKX2.5(+) cardiac homeobox protein Myocardial progenitor
cKIT(-) Mast/stem cell growth factor receptor (SCFR), also known as proto-oncogene c-Kit or tyrosine-protein kinase Kit or CD117 Cardiac mesoderm
cTNT(-) cardiac troponin-T Nascent cardiomyocytes
PDGFR-a platelet-derived growth factor receptor alpha Cardiac mesoderm
MESP1 mesoderm posterior 1 Cardiac mesoderm
ISL1 Islet 1 Myocardial progenitor

References:

  1. iCell Cardiac Progenitor Cells Datasheet
  2. iCell Cardiac Progenitor Cells Prototype User’s Guide
  3. Application Protocol: Modeling Cardiomyocyte Differentiation: Wnt- and Activin/TGFβ-inhibitor Induction with Flow Cytometry Analysis
  4. Application Protocol: Modeling Cardiac Proliferation: bFGF Induction with High Content Analysis
  5. Kattman poster: Robust and Scalable Production of Multipotent Cardiac Progenitor Cells from Human Induced Pluripotent Stem Cells and Their Use in Proliferation and Differentiation Screens
  6. Drowley et al (2016) Human Induced Pluripotent Stem Cell-Derived Cardiac Progenitor Cells in Phenotypic Screening: A Transforming Growth Factor-β Type 1 Receptor Kinase Inhibitor Induces Efficient Cardiac Differentiation