Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Alerts
  • Advertising
  • Job board
  • Subscribe
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Author's Takes
  • Reviews
    • View all reviews ...
    • Lung inflammatory injury and tissue repair (Jul 2023)
    • Immune Environment in Glioblastoma (Feb 2023)
    • Korsmeyer Award 25th Anniversary Collection (Jan 2023)
    • Aging (Jul 2022)
    • Next-Generation Sequencing in Medicine (Jun 2022)
    • New Therapeutic Targets in Cardiovascular Diseases (Mar 2022)
    • Immunometabolism (Jan 2022)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Commentaries
    • Research letters
    • Letters to the editor
    • Editorials
    • Viewpoint
    • Top read articles
  • Clinical Medicine
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Author's Takes
  • In-Press Preview
  • Commentaries
  • Research letters
  • Letters to the editor
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Alerts
  • Advertising
  • Job board
  • Subscribe
  • Contact
Vascular niche promotes hematopoietic multipotent progenitor formation from pluripotent stem cells
Jennifer L. Gori, … , Shahin Rafii, Hans-Peter Kiem
Jennifer L. Gori, … , Shahin Rafii, Hans-Peter Kiem
Published February 9, 2015
Citation Information: J Clin Invest. 2015;125(3):1243-1254. https://doi.org/10.1172/JCI79328.
View: Text | PDF
Technical Advance Stem cells

Vascular niche promotes hematopoietic multipotent progenitor formation from pluripotent stem cells

  • Text
  • PDF
Abstract

Pluripotent stem cells (PSCs) represent an alternative hematopoietic stem cell (HSC) source for treating hematopoietic disease. The limited engraftment of human PSC–derived (hPSC-derived) multipotent progenitor cells (MPP) has hampered the clinical application of these cells and suggests that MPP require additional cues for definitive hematopoiesis. We hypothesized that the presence of a vascular niche that produces Notch ligands jagged-1 (JAG1) and delta-like ligand-4 (DLL4) drives definitive hematopoiesis. We differentiated hes2 human embryonic stem cells (hESC) and Macaca nemestrina–induced PSC (iPSC) line-7 with cytokines in the presence or absence of endothelial cells (ECs) that express JAG1 and DLL4. Cells cocultured with ECs generated substantially more CD34+CD45+ hematopoietic progenitors compared with cells cocultured without ECs or with ECs lacking JAG1 or DLL4. EC-induced cells exhibited Notch activation and expressed HSC-specific Notch targets RUNX1 and GATA2. EC-induced PSC-MPP engrafted at a markedly higher level in NOD/SCID/IL-2 receptor γ chain–null (NSG) mice compared with cytokine-induced cells, and low-dose chemotherapy-based selection further increased engraftment. Long-term engraftment and the myeloid-to-lymphoid ratio achieved with vascular niche induction were similar to levels achieved for cord blood–derived MPP and up to 20-fold higher than those achieved with hPSC-derived MPP engraftment. Our findings indicate that endothelial Notch ligands promote PSC-definitive hematopoiesis and production of long-term engrafting CD34+ cells, suggesting these ligands are critical for HSC emergence.

Authors

Jennifer L. Gori, Jason M. Butler, Yan-Yi Chan, Devikha Chandrasekaran, Michael G. Poulos, Michael Ginsberg, Daniel J. Nolan, Olivier Elemento, Brent L. Wood, Jennifer E. Adair, Shahin Rafii, Hans-Peter Kiem

×

Figure 1

Endothelial Notch ligands regulate emergence and expansion of hematopoietic progenitor cells from PSCs.

Options: View larger image (or click on image) Download as PowerPoint
Endothelial Notch ligands regulate emergence and expansion of hematopoie...
(A) Left: CD34+CD45+ cell yield per million input of cells ± ECs and Notch ligand– replete and –depleted conditions. Right: fold change in CD34+CD45+ cells relative to MPP generated in cytokines. JAG1-KD, ECs transduced with shRNA to JAG1; DLL4-KD, ECs transduced with shRNA to DLL4, –, cytokines alone. (B) Notch signaling target expression on day 15. Left: MniPSC-7–MPP; right: day hes2-MPP after coculture in indicated conditions. Levels calibrated to expression in PSC-MPP cocultured with WT ECs and normalized to β-actin. (C) Number of hematopoietic CFUs from day-15 MniPSC-7–MPP after expansion ± WT ECs. Right: Mn BM CD34+ activity (control). CFUs per 100,000 cells plated. Plating efficiency noted in parentheses. Far right: CFUs from day 15 MniPSC-7–MPP and BM CD34+ cells. Original magnification, ×20. (D) Primate CD45+ cells in blood 12 weeks after transplantation (1 transplant experiment, n = 3 mice/group, bars represent mean/group). **P < 0.005; ***P < 0.0005, Student’s t test. Differentiation studies ± Notch ligand–depleted ECs were conducted in 2 MniPSC lines and 1 hESC line (hes2) in 3 independent experiments per cell line. Differentiation studies comparing induction with cytokines alone and WT ECs were conducted in 2 MniPSC lines and 1 hESC line in 6 independent experiments per cell line.

Copyright © 2023 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts