Isolation of an ES-Derived Cardiovascular Multipotent Cell Population Based on VE-Cadherin Promoter Activity

Author:

Maltabe Violetta A.12,Barka Eleonora3,Kontonika Marianthi45,Florou Dimitra1,Kouvara-Pritsouli Maria1,Roumpi Maria3,Agathopoulos Simeon3,Kolettis Theofilos M.45,Kouklis Panos12ORCID

Affiliation:

1. Laboratory of Biology, Medical School, University of Ioannina, Ioannina, Greece

2. Department of Biomedical Research, Institute of Molecular Biology & Biotechnology, Foundation of Research and Technology-Hellas, University Campus, 45110 Ioannina, Greece

3. Ceramics and Composites Laboratory Department of Materials Science and Engineering, University of Ioannina, Ioannina, Greece

4. Department of Cardiology, University of Ioannina Medical School, Ioannina, Greece

5. Cardiovascular Research Institute, Ioannina, Greece

Abstract

Embryonic Stem (ES) or induced Pluripotent Stem (iPS) cells are important sources for cardiomyocyte generation, targeted for regenerative therapies. Several in vitro protocols are currently utilized for their differentiation, but the value of cell-based approaches remains unclear. Here, we characterized a cardiovascular progenitor population derived during ES differentiation, after selection based on VE-cadherin promoter (Pvec) activity. ESCs were genetically modified with an episomal vector, allowing the expression of puromycin resistance gene, under Pvec activity. Puromycin-surviving cells displayed cardiac and endothelial progenitor cells characteristics. Expansion and self-renewal of this cardiac and endothelial dual-progenitor population (CEDP) were achieved by Wnt/β-catenin pathway activation. CEDPs express early cardiac developmental stage-specific markers but not markers of differentiated cardiomyocytes. Similarly, CEDPs express endothelial markers. However, CEDPs can undergo differentiation predominantly to cTnT+(~47%) and VE-cadherin+(~28%) cells. Transplantation of CEDPs in the left heart ventricle of adult rats showed that CEDPs-derived cells survive and differentiate in vivo for at least 14 days after transplantation. A novel, dual-progenitor population was isolated during ESCs differentiation, based on Pvec activity. This lineage can self-renew, permitting its maintenance as a source of cardiovascular progenitor cells and constitutes a useful source for regenerative approaches.

Funder

European Social Fund

Publisher

Hindawi Limited

Subject

Cell Biology,Molecular Biology

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