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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / metabolism
  • Antigens, CD34 / metabolism
  • Cadherins / metabolism
  • Cell Differentiation
  • Cell Engineering / methods
  • Cell Line
  • Cell Lineage
  • Chemokine CXCL10 / metabolism
  • Culture Media, Serum-Free
  • Female
  • Hemangioblasts / cytology*
  • Hemangioblasts / metabolism*
  • Hemangioblasts / transplantation
  • Hematopoiesis
  • Hematopoietic Stem Cell Transplantation
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / metabolism*
  • Heterografts
  • Humans
  • Leukocyte Common Antigens / metabolism
  • Mice
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism*
  • Pluripotent Stem Cells / transplantation
  • Signal Transduction
  • Stem Cell Niche

Substances

  • Antigens, CD
  • Antigens, CD34
  • CXCL10 protein, human
  • Cadherins
  • Chemokine CXCL10
  • Culture Media, Serum-Free
  • cadherin 5
  • Leukocyte Common Antigens
  • PTPRC protein, human