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Neurotrophins mediate human embryonic stem cell survival

Nature Biotechnology volume 24, pages 344–350 (2006)Cite this article

Abstract

Growth of human embryonic stem (hES) cells as a pluripotent population requires a balance between survival, proliferation and self-renewal signals. Here we demonstrate that hES cells express receptors of the tropomyosin-related kinase (TRK) family, which mediate antiapoptotic signals. We show that three TRK ligands, brain-derived neurotrophic factor, neurotrophin 3 and neurotrophin 4, are survival factors for hES cells. Addition of neurotrophins to hES cell cultures effects a 36-fold improvement in their clonal survival. hES cell cultures maintained in medium containing neurotrophins remain diploid and retain full developmental potency. In the presence of neurotrophins, TRK receptors in hES cells are phosphorylated; TRK receptor inhibition leads to hES cell apoptosis. The survival activity of neurotrophins in hES cells is mediated by the phosphatidylinositol-3-kinase pathway but not the mitogen-activated protein kinase pathway. Neurotrophins improve hES cell survival and may facilitate their manipulation and the development of high-throughput screens to identify factors responsible for hES cell differentiation.

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Figure 1: Human embryonic stem (hES) cells express members of the TRK receptor tyrosine kinase family of neurotrophin receptors.
Figure 2: Neurotrophins are expressed by MEFs.
Figure 3: Disruption of neurotrophin signaling leads to TRK receptor dephosphorylation and hES cell death.
Figure 4: PI-3K activity is required for hES cell survival.

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References

  1. Thomson, J.A. & Odorico, J.S. Human embryonic stem cell and embryonic germ cell lines. Trends Biotechnol. 18, 53–57 (2000).

    Article  CAS  Google Scholar 

  2. Trounson, A. & Pera, M. Human embryonic stem cells. Fertil. Steril. 76, 660–661 (2001).

    Article  CAS  Google Scholar 

  3. Donovan, P.J. & Gearhart, J. The end of the beginning for pluripotent stem cells. Nature 414, 92–97 (2001).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  5. Amit, M. et al. Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev. Biol. 227, 271–278 (2000).

    Article  CAS  Google Scholar 

  6. Sato, N., Meijer, L., Skaltsounis, L., Greengard, P. & Brivanlou, A.H. Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat. Med. 10, 55–63 (2004).

    Article  CAS  Google Scholar 

  7. 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).

    Article  CAS  Google Scholar 

  8. Beattie, G.M. et al. Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells 23, 489–495 (2005).

    Article  CAS  Google Scholar 

  9. Sato, N. et al. Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev. Biol. 260, 404–413 (2003).

    Article  CAS  Google Scholar 

  10. Richards, M., Tan, S.P., Tan, J.H., Chan, W.K. & Bongso, A. The transcriptome profile of human embryonic stem cells as defined by SAGE. Stem Cells 22, 51–64 (2004).

    Article  CAS  Google Scholar 

  11. Pyle, A.D., Donovan, P.J. & Lock, L.F. Chipping away at 'stemness'. Genome Biol. 5, 235.1–235.2 (2004).

    Article  Google Scholar 

  12. Patapoutian, A. & Reichardt, L.F. Trk receptors: mediators of neurotrophin action. Curr. Opin. Neurobiol. 11, 272–280 (2001).

    Article  CAS  Google Scholar 

  13. Chao, M.V. Neurotrophins and their receptors: a convergence point for many signalling pathways. Nat. Rev. Neurosci. 4, 299–309 (2003).

    Article  CAS  Google Scholar 

  14. Li, X.J. et al. Specification of motoneurons from human embryonic stem cells. Nat. Biotechnol. 23, 215–221 (2005).

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  17. Berg, M.M., Sternberg, D.W., Parada, L.F. & Chao, M.V. K-252a inhibits nerve growth factor-induced trk proto-oncogene tyrosine phosphorylation and kinase activity. J. Biol. Chem. 267, 13–16 (1992).

    CAS  PubMed  Google Scholar 

  18. Bramson, H.N. et al. Oxindole-based inhibitors of cyclin-dependent kinase 2 (CDK2): design, synthesis, enzymatic activities, and X-ray crystallographic analysis. J. Med. Chem. 44, 4339–4358 (2001).

    Article  CAS  Google Scholar 

  19. Wood, E.R. et al. Discovery and in vitro evaluation of potent TrkA kinase inhibitors: oxindole and aza-oxindoles. Bioorg. Med. Chem. Lett. 14, 953–957 (2004).

    Article  CAS  Google Scholar 

  20. Daheron, L. et al. LIF/STAT3 signaling fails to maintain self-renewal of human embryonic stem cells. Stem Cells 22, 770–778 (2004).

    Article  CAS  Google Scholar 

  21. Humphrey, R.K. et al. Maintenance of pluripotency in human embryonic stem cells is STAT3 independent. Stem Cells 22, 522–530 (2004).

    Article  CAS  Google Scholar 

  22. Draper, J.S. et al. Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat. Biotechnol. 22, 53–54 (2004).

    Article  CAS  Google Scholar 

  23. Ginty, D.D. & Segal, R.A. Retrograde neurotrophin signaling: Trk-ing along the axon. Curr. Opin. Neurobiol. 12, 268–274 (2002).

    Article  CAS  Google Scholar 

  24. Seifer, D.B., Feng, B., Shelden, R.M., Chen, S. & Dreyfus, C.F. Brain-derived neurotrophic factor: a novel human ovarian follicular protein. J. Clin. Endocrinol. Metab. 87, 655–659 (2002).

    Article  CAS  Google Scholar 

  25. Resnick, J.L., Bixler, L.S., Cheng, L. & Donovan, P.J. Long-term proliferation of mouse primordial germ cells in culture. Nature 359, 550–551 (1992).

    Article  CAS  Google Scholar 

  26. Kaplan, D.R., Martin-Zanca, D. & Parada, L.F. Tyrosine phosphorylation and tyrosine kinase activity of the trk proto-oncogene product induced by NGF. Nature 350, 158–160 (1991).

    Article  CAS  Google Scholar 

  27. Dalal, R. & Djakiew, D. Molecular characterization of neurotrophin expression and the corresponding tropomyosin receptor kinases (trks) in epithelial and stromal cells of the human prostate. Mol. Cell. Endocrinol. 134, 15–22 (1997).

    Article  CAS  Google Scholar 

  28. Qian, X. & Ginty, D.D. SH2-B and APS are multimeric adapters that augment TrkA signaling. Mol. Cell. Biol. 21, 1613–1620 (2001).

    Article  CAS  Google Scholar 

  29. Gopalan, G., Chan, C.S. & Donovan, P.J. A novel mammalian, mitotic spindle-associated kinase is related to yeast and fly chromosome segregation regulators. J. Cell Biol. 138, 643–656 (1997).

    Article  CAS  Google Scholar 

  30. Stephens, R.M. et al. Trk receptors use redundant signal transduction pathways involving SHC and PLC-gamma 1 to mediate NGF responses. Neuron 12, 691–705 (1994).

    Article  CAS  Google Scholar 

  31. Shamblott, M.J. et al. Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc. Natl. Acad. Sci. USA 95, 13726–13731 (1998).

    Article  CAS  Google Scholar 

  32. Cowan, C.A. et al. Derivation of embryonic stem-cell lines from human blastocysts. N. Engl. J. Med. 350, 1353–1356 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are indebted to Linzhao Cheng, Ira Daar, Lino Tessarollo, Brad Harris, John Gearhart and Ian McNiece for helpful suggestions and comments; Lino Tessarollo, Gautam Dravid, David Ginty, Hongjun Song, Xin Duan, Pantelis Tsoulfas, Karen Lackey and Rasi Wickramsinghe for reagents and advice; and Gail Stetten and the Cytogenetics Facility for karyotype analysis. We are also grateful to Sean Donovan and Robert Clouse III for their support and patience and members of the Donovan Lab for their support and encouragement. L.F.L. and P.J.D. dedicate this work to Camilynn I. Brannan (1963–2002). This work was supported by a National Research Service Award (to A.D.P.) by a National Institutes of Health Grant (to P.J.D.) and by institutional funds from the Johns Hopkins University School of Medicine (to L.F.L. and P.J.D.).

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Author notes

  1. Leslie F Lock & Peter J Donovan

    Present address: Stem Cell Program and Departments of Developmental and Cell Biology and of Biological Chemistry, Hewitt Hall, University of California, Irvine, Irvine, California, 92697, USA

Authors and Affiliations

  1. Germ Cell and Stem Cell Group, Stem Cell Biology Program, Institute for Cell Engineering, Johns Hopkins University, School of Medicine, Broadway Research Building, Suite 759, 733 North Broadway, Baltimore, 21205, Maryland, USA

    April D Pyle & Peter J Donovan

  2. Genetic Models Group, Stem Cell Biology Program, Institute for Cell Engineering, Johns Hopkins University, School of Medicine, Broadway Research Building, Suite 759, 733 North Broadway, Baltimore, 21205, Maryland, USA

    Leslie F Lock

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Correspondence to Peter J Donovan.

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Supplementary information

Supplementary Fig. 1

Expression of neurotrophins receptors on H9 hES cells and controls for antibody reactivity. (PDF 701 kb)

Supplementary Fig. 2

Effect of neurotrophins on hES cell differentiation and TRK phosphorylation. (PDF 1602 kb)

Supplementary Fig. 3

Effect of neurotrophins on the population doubling time of hES cells. (PDF 875 kb)

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Pyle, A., Lock, L. & Donovan, P. Neurotrophins mediate human embryonic stem cell survival. Nat Biotechnol 24, 344–350 (2006). https://doi.org/10.1038/nbt1189

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