Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFΒ inhibition is Id1 dependent Journal Article
| Authors: | 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. |
| Article Title: | Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFΒ inhibition is Id1 dependent |
| 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. |
| Keywords: | signal transduction; controlled study; protein expression; human cell; nonhuman; flow cytometry; cell proliferation; mouse; cells, cultured; smooth muscle fiber; gene expression; cell growth; inhibitor of differentiation 1; transforming growth factor beta; embryonic stem cell; protein protein interaction; green fluorescent protein; animal experiment; cell differentiation; vasculotropin receptor 2; alpha smooth muscle actin; endothelium cell; endothelial cells; umbilical vein; transforming growth factors; stem cells; embryonic stem cells; 4 [4 (1,3 benzodioxol 5 yl) 5 (2 pyridinyl) 1h imidazol 2 yl]benzamide; inhibitor of differentiation protein 1; fluorescence activated cell sorting; clinical application; stem cell expansion; maintenance; vascular endothelial cadherin; monolayers; self assembly; ephrin b2; human embryonic stem cells; in-phase; vascular cells; vasculogenic; ve-cadherin; vegfr-2; blood vessel prostheses; fluorescent screens; yellow fluorescent protein |
| Journal Title: | Nature Biotechnology |
| Volume: | 28 |
| Issue: | 2 |
| ISSN: | 1087-0156 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2010-02-01 |
| Start Page: | 161 |
| End Page: | 166 |
| Language: | English |
| DOI: | 10.1038/nbt.1605 |
| PUBMED: | 20081865 |
| PROVIDER: | scopus |
| PMCID: | PMC2931334 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 10" - "Export Date: 20 April 2011" - "CODEN: NABIF" - "Source: Scopus" |
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