title: Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells.
abstract: Human endothelial cells (ECs) and pericytes are of great interest for research on vascular development and disease, as well as for 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 d), isolation (1 d) and 4 or 10 d of expansion of ECs and pericytes, respectively. We also describe two assays for functional evaluation of hPSC-derived ECs: (i) primary vascular plexus formation upon coculture 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.
authors.affiliation: 1] Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands. [2] Department of Molecular Cell Biology, Cancer Genomics Centre, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Valeria V
authors.initials: VV
authors.lastname: Orlova
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Francijna E
authors.initials: FE
authors.lastname: van den Hil
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Sandra
authors.initials: S
authors.lastname: Petrus-Reurer
authors.affiliation: Department of Molecular Cell Biology, Cancer Genomics Centre, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Yvette
authors.initials: Y
authors.lastname: Drabsch
authors.affiliation: Department of Molecular Cell Biology, Cancer Genomics Centre, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Ten Dijke
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Christine L
authors.initials: CL
authors.lastname: Mummery
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology; D064593:Heterografts / Q000098:blood supply / Q000166:cytology; D006801:Humans; D020286:Pericytes / Q000166:cytology* / Q000502:physiology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D015027:Zebrafish