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Nat Commun. 2017 May 25;8:15380. doi: 10.1038/ncomms15380.

Engineering the haemogenic niche mitigates endogenous inhibitory signals and controls pluripotent stem cell-derived blood emergence.

Author information

1
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada M5S 3ES.
2
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada M5S 3G9.
3
Biological Sciences, Sunnybrook Research Institute, Toronto, Ontario, Canada M4N 3M5.
4
Life Sciences (Biochemistry), University of Waterloo, Waterloo, Ontario, Canada N2L 3G1.
5
Collaborative Program in Developmental Biology, University of Toronto, Toronto, Ontario, Canada M5S 3E1.
6
Department of Immunology, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
7
Medicine by Design, a Canada First Research Excellence Program at the University of Toronto, Toronto, Ontario, Canada M5S 3G9.
8
Terrence Donnelly Centre for Cellular &Biomolecular Research, University of Toronto, Toronto, Ontario, Canada M5S 3E1.

Abstract

Efforts to recapitulate haematopoiesis, a process guided by spatial and temporal inductive signals, to generate haematopoietic progenitors from human pluripotent stem cells (hPSCs) have focused primarily on exogenous signalling pathway activation or inhibition. Here we show haemogenic niches can be engineered using microfabrication strategies by micropatterning hPSC-derived haemogenic endothelial (HE) cells into spatially-organized, size-controlled colonies. CD34+VECAD+ HE cells were generated with multi-lineage potential in serum-free conditions and cultured as size-specific haemogenic niches that displayed enhanced blood cell induction over non-micropatterned cultures. Intra-colony analysis revealed radial organization of CD34 and VECAD expression levels, with CD45+ blood cells emerging primarily from the colony centroid area. We identify the induced interferon gamma protein (IP-10)/p-38 MAPK signalling pathway as the mechanism for haematopoietic inhibition in our culture system. Our results highlight the role of spatial organization in hPSC-derived blood generation, and provide a quantitative platform for interrogating molecular pathways that regulate human haematopoiesis.

PMID:
28541275
PMCID:
PMC5477512
DOI:
10.1038/ncomms15380
[Indexed for MEDLINE]
Free PMC Article

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