RPE and Stem Cell Therapy

acceptedVersionPeer reviewe

Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells

Stem cells & developmental biolog

Differentiation and functionality of endothelial cells and pericytes from human induced pluripotent stem cells

Stem cells & developmental biolog

Functionality of Endothelial Cells and Pericytes From Human Pluripotent Stem Cells Demonstrated in Cultured Vascular Plexus and Zebrafish Xenografts

Stem cells & developmental biolog

Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells

Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease as well as future therapy. This protocol describes the efficient generation of ECs and pericytes from human pluripotent stem cells (hPSCs) under defined conditions. Essential steps for hPSC culture, differentiation, isolation and functional characterization of ECs and pericytes are described. Substantial numbers of both cell types can be derived in only 2-3 weeks: this involves differentiation (10 days), isolation (1 day) and 4 or 10 days expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon co-culture with hPSC-derived pericytes and (ii) incorporation in the vasculature of zebrafish xenografts in vivo. These assays can be used to test the quality and drug sensitivity of hPSC-derived ECs and model vascular diseases with patient-derived hPSCs