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IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro

Abstract

Distinctive properties of stem cells are not autonomously achieved, and recent evidence points to a level of external control from the microenvironment. Here, we demonstrate that self-renewal and pluripotent properties of human embryonic stem (ES) cells depend on a dynamic interplay between human ES cells and autologously derived human ES cell fibroblast-like cells (hdFs). Human ES cells and hdFs are uniquely defined by insulin-like growth factor (IGF)- and fibroblast growth factor (FGF)-dependence. IGF 1 receptor (IGF1R) expression was exclusive to the human ES cells, whereas FGF receptor 1 (FGFR1) expression was restricted to surrounding hdFs. Blocking the IGF-II/IGF1R pathway reduced survival and clonogenicity of human ES cells, whereas inhibition of the FGF pathway indirectly caused differentiation. IGF-II is expressed by hdFs in response to FGF, and alone was sufficient in maintaining human ES cell cultures. Our study demonstrates a direct role of the IGF-II/IGF1R axis on human ES cell physiology and establishes that hdFs produced by human ES cells themselves define the stem cell niche of pluripotent human stem cells.

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Figure 1: Receptor expression reveals human ES cell heterogeneity.
Figure 2: Immunocytochemistry staining of undifferentiated human ES cell cultures.
Figure 3: Differential IGF and FGF effects on human ES cells.
Figure 4: Function of IGF and its production in human ES cell cultures.
Figure 5: FGF-induced TGF-β signals required for human ES cell pluripotency.
Figure 6: Proposed model of human ES cell paracrine regulation.

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Acknowledgements

S.C.B. is supported by a CIHR Canada Graduate Scholarship doctoral award; M.H.S. by a postgraduate scholarship award from the Stem Cell Network and CIHR Canada Graduate Scholarship doctoral award; and M.Bh. by the Canadian Chair Program who holds the Canada Research Chair in human stem cell biology and Michael G. DeGroote Chair in Stem Cell Biology. This work was supported by a grant from the Ontario Research and Development Challenge Fund (ORDCF) to G.L. and by CIHR and NCIC to M.Bh. We also are grateful for the help of L. Gallacher and R. Mondeh with culture assistance, the Krembil Centre at the Robarts and M. Sibly and J. Trowbridge for useful suggestions, and A. Nagy, J. Rossant, M. Gertsenstein, K. Vinterstein, M. Mileikovsky and J. Draper for providing the CA1 human ES cell line.

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Correspondence to Mickie Bhatia.

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Bendall, S., Stewart, M., Menendez, P. et al. IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro. Nature 448, 1015–1021 (2007). https://doi.org/10.1038/nature06027

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