Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFΒ inhibition is Id1 dependent

Daylon James, Hyung Song Nam, Marco Seandel, Daniel Nolan, Tyler Janovitz, Mark Tomishima, Lorenz Studer, Gabsang Lee, David Lyden, Robert Benezra, Nikica Zaninovic, Zev Rosenwaks, Sina Y. Rabbany, Shahin Rafii

Research output: Contribution to journalArticle

Abstract

Previous efforts to differentiate human embryonic stem cells (hESCs) into endothelial cells have not achieved sustained expansion and stability of vascular cells. To define vasculogenic developmental pathways and enhance differentiation, we used an endothelial cell-specific VE-cadherin promoter driving green fluorescent protein (GFP) (hVPr-GFP) to screen for factors that promote vascular commitment. In phase 1 of our method, inhibition of transforming growth factor (TGF)Β at day 7 of differentiation increases hVPr-GFP + cells by tenfold. In phase 2, TGFΒ inhibition maintains the proliferation and vascular identity of purified endothelial cells, resulting in a net 36-fold expansion of endothelial cells in homogenous monolayers, which exhibited a transcriptional profile of Id1 high VEGFR2 high VE-cadherin + ephrinB2 +. Using an Id1-YFP hESC reporter line, we showed that TGFΒ inhibition sustains Id1 expression in hESC-derived endothelial cells and that Id1 is required for increased proliferation and preservation of endothelial cell commitment. Our approach provides a serum-free method for differentiation and long-term maintenance of hESC-derived endothelial cells at a scale relevant to clinical application.

Original languageEnglish (US)
Pages (from-to)161-166
Number of pages6
JournalNature biotechnology
Volume28
Issue number2
DOIs
StatePublished - Feb 1 2010
Externally publishedYes

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ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

James, D., Nam, H. S., Seandel, M., Nolan, D., Janovitz, T., Tomishima, M., Studer, L., Lee, G., Lyden, D., Benezra, R., Zaninovic, N., Rosenwaks, Z., Rabbany, S. Y., & Rafii, S. (2010). Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFΒ inhibition is Id1 dependent. Nature biotechnology, 28(2), 161-166. https://doi.org/10.1038/nbt.1605