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Abstract
NOTCH signaling is required for the arterial specification and formation of hematopoietic stem cells (HSCs) and lympho-myeloid progenitors in the embryonic aorta-gonad-mesonephros region and extraembryonic vasculature from a distinct lineage of vascular endothelial cells with hemogenic potential. However, the role of NOTCH signaling in hemogenic endothelium (HE) specification from human pluripotent stem cell (hPSC) has not been studied. Here, using a chemically defined hPSC differentiation system combined with the use of DLL1-Fc and DAPT to manipulate NOTCH, we discover that NOTCH activation in hPSC-derived immature HE progenitors leads to formation of CD144+CD43−CD73−DLL4+Runx1 + 23-GFP+ arterial-type HE, which requires NOTCH signaling to undergo endothelial-to-hematopoietic transition and produce definitive lympho-myeloid and erythroid cells. These findings demonstrate that NOTCH-mediated arterialization of HE is an essential prerequisite for establishing definitive lympho-myeloid program and suggest that exploring molecular pathways that lead to arterial specification may aid in vitro approaches to enhance definitive hematopoiesis from hPSCs.
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1 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA
2 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI, USA
3 Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA; Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
4 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI, USA; Department of Biomedical Sciences, University of Wisconsin School of Veterinary Medicine, Madison, WI, USA
5 Morgridge Institute for Research, Madison, WI, USA
6 Department of Pharmacology, University of Illinois, Chicago, IL, USA
7 Stem Cell Program and Division of Hematology/Oncology, Children’s Hospital Boston, Harvard Medical School and Howard Hughes Medical Institute, Boston, MA, USA
8 Morgridge Institute for Research, Madison, WI, USA; Department of Cell and Regenerative Biology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA; Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, CA, USA
9 Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA; Department of Cell and Regenerative Biology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA