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

Valeria V. Orlova*, Francijna E. van den Hil, Sandra Petrus-Reurer, Yvette Drabsch, Peter ten Dijke, Christine L. Mummery

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)1514-1531
Number of pages18
JournalNature Protocols
Volume9
Issue number6
DOIs
Publication statusPublished - Jun 2014

Keywords

  • MESENCHYMAL STEM
  • PROGENITOR CELLS
  • DRUG DISCOVERY
  • CORD BLOOD
  • ZEBRAFISH
  • DIFFERENTIATION
  • DERIVATION
  • INHIBITION
  • DISEASE
  • CANCER

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