Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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


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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

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.


  1. 1

    Scadden, D. T. The stem-cell niche as an entity of action. Nature 441, 1075–1079 (2006)

    ADS  CAS  Article  Google Scholar 

  2. 2

    Thomson, J. A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147 (1998)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Xu, C. et al. Feeder-free growth of undifferentiated human embryonic stem cells. Nature Biotechnol. 19, 971–974 (2001)

    CAS  Article  Google Scholar 

  4. 4

    Ludwig, T. E. et al. Derivation of human embryonic stem cells in defined conditions. Nature Biotechnol. 24, 185–187 (2006)

    CAS  Article  Google Scholar 

  5. 5

    Hoffman, L. M. & Carpenter, M. K. Characterization and culture of human embryonic stem cells. Nature Biotechnol. 23, 699–708 (2005)

    CAS  Article  Google Scholar 

  6. 6

    Stewart, M. H. et al. Clonal isolation of hESCs reveals heterogeneity within the pluripotent stem cell compartment. Nature Methods 3, 807–815 (2006)

    CAS  Article  Google Scholar 

  7. 7

    Ying, Q. L., Nichols, J., Chambers, I. & Smith, A. BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell 115, 281–292 (2003)

    CAS  Article  Google Scholar 

  8. 8

    Dechiara, T. M., Efstratiadis, A. & Robertson, E. J. A growth-deficiency phenotype in heterozygous mice carrying an insulin-like growth factor II gene disrupted by targeting. Nature 345, 78–80 (1990)

    ADS  CAS  Article  Google Scholar 

  9. 9

    Kauma, S. W. Cytokines in implantation. J. Reprod. Fertil. Suppl. 55, 31–42 (2000)

  10. 10

    Heyner, S. Growth factors in preimplantation development: role of insulin and insulin-like growth factors. Early Pregnancy 3, 153–163 (1997)

    CAS  PubMed  Google Scholar 

  11. 11

    Sperger, J. M. et al. Gene expression patterns in human embryonic stem cells and human pluripotent germ cell tumors. Proc. Natl Acad. Sci. USA 100, 13350–13355 (2003)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Levenstein, M. E. et al. Basic fibroblast growth factor support of human embryonic stem cell self-renewal. Stem Cells 24, 568–574 (2006)

    CAS  Article  Google Scholar 

  13. 13

    Xu, R. H. et al. Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. Nature Methods 2, 185–190 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Dvorak, P. et al. Expression and potential role of fibroblast growth factor 2 and its receptors in human embryonic stem cells. Stem Cells 23, 1200–1211 (2005)

    CAS  Article  Google Scholar 

  15. 15

    Xu, C. et al. Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. Stem Cells 23, 315–323 (2005)

    CAS  Article  Google Scholar 

  16. 16

    Rosler, E. S. et al. Long-term culture of human embryonic stem cells in feeder-free conditions. Dev. Dyn. 229, 259–274 (2004)

    CAS  Article  Google Scholar 

  17. 17

    Baserga, R., Peruzzi, F. & Reiss, K. The IGF-1 receptor in cancer biology. Int. J. Cancer 107, 873–877 (2003)

    CAS  Article  Google Scholar 

  18. 18

    Hofmann, F. & Garcia-Echeverria, C. Blocking the insulin-like growth factor-I receptor as a strategy for targeting cancer. Drug Discov. Today 10, 1041–1047 (2005)

    CAS  Article  Google Scholar 

  19. 19

    Tropepe, V. et al. Direct neural fate specification from embryonic stem cells: a primitive mammalian neural stem cell stage acquired through a default mechanism. Neuron 30, 65–78 (2001)

    CAS  Article  Google Scholar 

  20. 20

    Vallier, L., Reynolds, D. & Pedersen, R. A. Nodal inhibits differentiation of human embryonic stem cells along the neuroectodermal default pathway. Dev. Biol. 275, 403–421 (2004)

    CAS  Article  Google Scholar 

  21. 21

    Wang, L., Li, L., Menendez, P., Cerdan, C. & Bhatia, M. Human embryonic stem cells maintained in the absence of mouse embryonic fibroblasts or conditioned media are capable of hematopoietic development. Blood 105, 4598–4603 (2005)

    CAS  Article  Google Scholar 

  22. 22

    Greber, B., Lehrach, H. & Adjaye, J. Fibroblast growth factor 2 transforming growth factor β signaling in mouse embryonic fibroblasts and human ESCs (hESCs) to support hESC self-renewal. Stem Cells 25, 455–464 (2007)

    CAS  Article  Google Scholar 

  23. 23

    Huang, S. & Terstappen, L. W. Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells. Nature 360, 745–749 (1992)

    ADS  CAS  Article  Google Scholar 

  24. 24

    Huang, S. & Terstappen, L. W. Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells. Nature 368, 664 (1994)

    ADS  CAS  Article  Google Scholar 

  25. 25

    Lighten, A. D., Hardy, K., Winston, R. M. & Moore, G. E. Expression of mRNA for the insulin-like growth factors and their receptors in human preimplantation embryos. Mol. Reprod. Dev. 47, 134–139 (1997)

    CAS  Article  Google Scholar 

  26. 26

    Chi, M. M., Schlein, A. L. & Moley, K. H. High insulin-like growth factor 1 (IGF-1) and insulin concentrations trigger apoptosis in the mouse blastocyst via down-regulation of the IGF-1 receptor. Endocrinology 141, 4784–4792 (2000)

    CAS  Article  Google Scholar 

  27. 27

    Mohammadi, M. et al. Structures of the tyrosine kinase domain of fibroblast growth factor receptor in complex with inhibitors. Science 276, 955–960 (1997)

    CAS  Article  Google Scholar 

  28. 28

    Inman, G. J. et al. SB-431542 is a potent and specific inhibitor of transforming growth factor-β superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol. Pharmacol. 62, 65–74 (2002)

    CAS  Article  Google Scholar 

  29. 29

    Laping, N. J. et al. Inhibition of transforming growth factor (TGF)-β1-induced extracellular matrix with a novel inhibitor of the TGF-β type I receptor kinase activity: SB-431542. Mol. Pharmacol. 62, 58–64 (2002)

    CAS  Article  Google Scholar 

  30. 30

    Orfao, A. et al. Flow cytometry in the diagnosis of cancer. Scand. J. Clin. Lab. Invest. Suppl. 221145–152 (1995)

  31. 31

    Orfao, A. et al. Flow cytometry: its applications in hematology. Haematologica 80, 69–81 (1995)

    CAS  PubMed  Google Scholar 

Download references


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.

Author information



Corresponding author

Correspondence to Mickie Bhatia.

Ethics declarations

Competing interests

Reprints and permissions information is available at The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-7 with Legends and Supplementary Tables 1-2. (PDF 9774 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing