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

TitleExpansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFbeta inhibition is Id1 dependent.
Publication TypeJournal Article
Year of Publication2010
AuthorsJames D, Nam H-song, Seandel M, Nolan D, Janovitz T, Tomishima M, Studer L, Lee G, Lyden D, Benezra R, Zaninovic N, Rosenwaks Z, Rabbany SY, Rafii S
JournalNat Biotechnol
Volume28
Issue2
Pagination161-6
Date Published2010 Feb
ISSN1546-1696
KeywordsCell Differentiation, Cells, Cultured, Embryonic Stem Cells, Endothelial Cells, Humans, Inhibitor of Differentiation Protein 1, Signal Transduction, Transforming Growth Factor beta
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)beta at day 7 of differentiation increases hVPr-GFP(+) cells by tenfold. In phase 2, TGFbeta 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 TGFbeta 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.

DOI10.1038/nbt.1605
Alternate JournalNat. Biotechnol.
PubMed ID20081865
PubMed Central IDPMC2931334
Grant ListP01 HL059312 / HL / NHLBI NIH HHS / United States
P01 HL059312-060006 / HL / NHLBI NIH HHS / United States
P01 HL059312-070006 / HL / NHLBI NIH HHS / United States
P01 HL059312-080006 / HL / NHLBI NIH HHS / United States
P01 HL059312-090006 / HL / NHLBI NIH HHS / United States
P01 HL059312-100006 / HL / NHLBI NIH HHS / United States
P01 HL067839 / HL / NHLBI NIH HHS / United States
P01 HL067839-010004 / HL / NHLBI NIH HHS / United States
P01 HL067839-020004 / HL / NHLBI NIH HHS / United States
P01 HL067839-030004 / HL / NHLBI NIH HHS / United States
P01 HL067839-040004 / HL / NHLBI NIH HHS / United States
P01 HL067839-050004 / HL / NHLBI NIH HHS / United States
P50 HL084936 / HL / NHLBI NIH HHS / United States
P50 HL084936-010003 / HL / NHLBI NIH HHS / United States
P50 HL084936-020003 / HL / NHLBI NIH HHS / United States
P50 HL084936-030003 / HL / NHLBI NIH HHS / United States
P50 HL084936-040003 / HL / NHLBI NIH HHS / United States
R01 DK095039 / DK / NIDDK NIH HHS / United States
R01 HL058707 / HL / NHLBI NIH HHS / United States
R01 HL058707-03 / HL / NHLBI NIH HHS / United States
R01 HL058707-04 / HL / NHLBI NIH HHS / United States
R01 HL061849 / HL / NHLBI NIH HHS / United States
R01 HL061849-02 / HL / NHLBI NIH HHS / United States
R01 HL061849-03 / HL / NHLBI NIH HHS / United States
R01 HL061849-03S1 / HL / NHLBI NIH HHS / United States
R01 HL061849-04 / HL / NHLBI NIH HHS / United States
R01 HL061849-05 / HL / NHLBI NIH HHS / United States
R01 HL075234 / HL / NHLBI NIH HHS / United States
R01 HL075234 / HL / NHLBI NIH HHS / United States
R01 HL075234-01 / HL / NHLBI NIH HHS / United States
R01 HL075234-02 / HL / NHLBI NIH HHS / United States
R01 HL075234-03 / HL / NHLBI NIH HHS / United States
R01 HL075234-04 / HL / NHLBI NIH HHS / United States
R01 HL097797 / HL / NHLBI NIH HHS / United States
R01 HL097797 / HL / NHLBI NIH HHS / United States
R01 HL097797-01 / HL / NHLBI NIH HHS / United States
R01 HL097797-02 / HL / NHLBI NIH HHS / United States
R01 HL097797-03 / HL / NHLBI NIH HHS / United States
R01 HL119872 / HL / NHLBI NIH HHS / United States
R21 HL083222 / HL / NHLBI NIH HHS / United States
R21 HL083222-01 / HL / NHLBI NIH HHS / United States
R21 HL083222-02 / HL / NHLBI NIH HHS / United States
RC1 AI080309 / AI / NIAID NIH HHS / United States
RC1 AI080309-01 / AI / NIAID NIH HHS / United States
RC2 HL101846 / HL / NHLBI NIH HHS / United States
T32 GM007739 / GM / NIGMS NIH HHS / United States
U01 HL066952 / HL / NHLBI NIH HHS / United States
U01 HL066952-010002 / HL / NHLBI NIH HHS / United States
U01 HL066952-020002 / HL / NHLBI NIH HHS / United States
U01 HL066952-030002 / HL / NHLBI NIH HHS / United States
U01 HL066952-040002 / HL / NHLBI NIH HHS / United States
U01 HL066952-050002 / HL / NHLBI NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
/ / Howard Hughes Medical Institute / United States