Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells
Open accessAbstract
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.
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HUVECs have exhibited poor engraftment, limited capacity to form vessels, and an inability to survive long-term after in vivo application.
“Additionally, HUVECs have exhibited poor engraftment, limited capacity to form vessels, and an inability to survive long-term after in vivo application [237,238].”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [238]
Human iPSC-derived ECs can form blood vessels and anastomose with the host vasculature when injected into a zebrafish model.
“Human iPSC-derived ECs can form blood vessels and anastomose with the host vasculature when injected into a zebrafish model [238].”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [238]
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“Human iPSC-derived ECs can form blood vessels and anastomose with the host vasculature when injected into a zebrafish model [238].”
BackgroundInfluential
- Authors
- Orlova, V. V., van den Hil, F. E., Petrus-Reurer, S., Drabsch, Y., ten Dijke, P., & Mummery, C. L.
- Affiliations
- Leiden University Medical Center (NL)
- Corresponding author
- Orlova, Valeria V
- Mummery, Christine L
- Venue
- Nature Protocols
- Published
- 2014-05-29
- Volume
- 9
- Issue
- 6
- Pages
- 1514-1531
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W1973626590
- Cited by
- 365
- References
- 5
- Citations by year
- 2026: 25 · 2025: 30 · 2024: 35 · 2023: 33 · 2022: 26 · 2021: 42 · 2020: 38 · 2019: 40 · 2018: 32 · 2017: 27 · 2016: 19 · 2015: 16 · 2014: 2
- FWCI
- 8.74
- Type
- journal article
- Language
- en
- Open access
- green
- zenodo.org
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Leiden Repository (Leiden University) (submittedVersion)
- Zenodo (CERN European Organization for Nuclear Research) (submittedVersion, cc-by)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Zebrafish Biomedical Research Applications
- Keywords
- endothelial cells, pericytes, human pluripotent stem cells, cell differentiation, zebrafish xenograft, vasculogenesis, vascular development, patient-derived stem cells, drug susceptibility testing
- MeSH terms
- Animals, Cell Differentiation, Humans, Zebrafish, Cell Culture Techniques, Pericytes, Pluripotent Stem Cells, Endothelial Cells, Cell Proliferation, Heterografts
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