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Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells

Nature Biotechnology volume 32, pages 554–561 (2014)Cite this article

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Abstract

Efforts to derive hematopoietic stem cells (HSCs) from human pluripotent stem cells (hPSCs) are complicated by the fact that embryonic hematopoiesis consists of two programs, primitive and definitive, that differ in developmental potential. As only definitive hematopoiesis generates HSCs, understanding how this program develops is essential for being able to produce this cell population in vitro. Here we show that both hematopoietic programs transition through hemogenic endothelial intermediates and develop from KDR+CD34−CD144− progenitors that are distinguished by CD235a expression. Generation of primitive progenitors (KDR+CD235a+) depends on stage-specific activin-nodal signaling and inhibition of the Wnt–β-catenin pathway, whereas specification of definitive progenitors (KDR+CD235a−) requires Wnt–β-catenin signaling during this same time frame. Together, these findings establish simple selective differentiation strategies for the generation of primitive or definitive hematopoietic progenitors by Wnt–β-catenin manipulation, and in doing so provide access to enriched populations for future studies on hPSC-derived hematopoietic development.

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Figure 1: Primitive hematopoiesis originates from a KDR+CD235a+ progenitor.
Figure 2: KDR+CD235a− mesoderm–derived CD34+CD43− cells possess definitive hematopoietic potential, but both CD34+ populations possess hemogenic endothelium-like potential.
Figure 3: Canonical Wnt signaling specifies definitive hematopoiesis.
Figure 4: Wnt manipulation allows for selective differentiation of primitive or definitive hematopoietic progenitors.
Figure 5: Model of primitive and definitive hematopoietic specification from hPSCs.

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Acknowledgements

We would like to thank the SickKids–UHN Flow Cytometry Facility for their expert assistance with cell sorting, in particular A. Khandani. This work was supported by National Institutes of Health grants U01 HL100395 and CIHR MOP93569, HOP83070 and MOP12927. C.M.S. and A.D. were supported by the McMurrich Post-Doctoral Fellowship and the Magna-Golftown Post-Doctoral Fellowship, respectively.

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  1. Christopher M Sturgeon

    Present address: Present address: Department of Internal Medicine, Hematology Division, Washington University School of Medicine, Saint Louis, Missouri, USA.,

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  1. McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada

    Christopher M Sturgeon, Andrea Ditadi, Geneve Awong, Marion Kennedy & Gordon Keller

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Contributions

C.M.S., A.D., G.A., M.K. and G.K. all participated in the design of the experiments. C.M.S., A.D., G.A. and M.K. performed the experiments. C.M.S. and G.K. wrote the manuscript.

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Correspondence to Gordon Keller.

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Sturgeon, C., Ditadi, A., Awong, G. et al. Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells. Nat Biotechnol 32, 554–561 (2014). https://doi.org/10.1038/nbt.2915

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