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Nanog promotes transfer of pluripotency after cell fusion

Abstract

Through cell fusion, embryonic stem (ES) cells can erase the developmental programming of differentiated cell nuclei and impose pluripotency1,2. Molecules that mediate this conversion should be identifiable in ES cells. One candidate is the variant homeodomain protein Nanog, which has the capacity to entrain undifferentiated ES cell propagation3,4. Here we report that in fusions between ES cells and neural stem (NS) cells, increased levels of Nanog stimulate pluripotent gene activation from the somatic cell genome and enable an up to 200-fold increase in the recovery of hybrid colonies, all of which show ES cell characteristics. Nanog also improves hybrid yield when thymocytes or fibroblasts are fused to ES cells; however, fewer colonies are obtained than from ES × NS cell fusions, consistent with a hierarchical susceptibility to reprogramming among somatic cell types. Notably, for NS × ES cell fusions elevated Nanog enables primary hybrids to develop into ES cell colonies with identical frequency to homotypic ES × ES fusion products. This means that in hybrids, increased Nanog is sufficient for the NS cell epigenome to be reset completely to a state of pluripotency. We conclude that Nanog can orchestrate ES cell machinery to instate pluripotency with an efficiency of up to 100% depending on the differentiation status of the somatic cell.

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Figure 1: Elevated levels of Nanog enhances formation of hybrid colonies from ES × NS cell fusions.
Figure 2: Characterization of ES × NS cell hybrids.
Figure 3: FACS purification and analysis of primary fusion products.
Figure 4: Nanog expression in NS cells increases the frequency of hybrid colonies.

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References

  1. Tada, M., Takahama, Y., Abe, K., Nakatsuji, N. & Tada, T. Nuclear reprogramming of somatic cells by in vitro hybridization with ES cells. Curr. Biol. 11, 1553–1558 (2001)

    Article  CAS  PubMed  Google Scholar 

  2. Ying, Q. L., Nichols, J., Evans, E. P. & Smith, A. G. Changing potency by spontaneous fusion. Nature 416, 545–548 (2002)

    Article  ADS  CAS  PubMed  Google Scholar 

  3. Chambers, I. et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655 (2003)

    Article  CAS  PubMed  Google Scholar 

  4. Mitsui, K. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642 (2003)

    Article  CAS  PubMed  Google Scholar 

  5. Campbell, K. H. S., McWhir, J., Ritchie, W. A. & Wilmut, I. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–66 (1996)

    Article  ADS  CAS  PubMed  Google Scholar 

  6. Cowan, C. A., Atienza, J., Melton, D. A. & Eggan, K. Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells. Science 309, 1369–1373 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  7. Kimura, H., Tada, M., Nakatsuji, N. & Tada, T. Histone code modifications on pluripotential nuclei of reprogrammed somatic cells. Mol. Cell. Biol. 24, 5710–5720 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Matveeva, N. M. et al. In vitro and in vivo study of pluripotency in intraspecific hybrid cells obtained by fusion of murine embryonic stem cells with splenocytes. Mol. Reprod. Dev. 50, 128–138 (1998)

    Article  CAS  PubMed  Google Scholar 

  9. Miller, R. A. & Ruddle, F. H. Teratocarcinoma × friend erythroleukemia cell hybrids resemble their pluripotent embryonal carcinoma parent. Dev. Biol. 56, 157–173 (1977)

    Article  CAS  PubMed  Google Scholar 

  10. Tada, M., Tada, T., Lefebvre, L., Barton, S. C. & Surani, M. A. Embryonic germ cells induce epigenetic reprogramming of somatic nucleus in hybrid cells. EMBO J. 16, 6510–6520 (1997)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Miller, R. A. & Ruddle, F. H. Pluripotent teratocarcinoma–thymus somatic cell hybrids. Cell 9, 45–55 (1976)

    Article  CAS  PubMed  Google Scholar 

  12. Conti, L. et al. Niche-independent symmetrical self-renewal of a mammalian tissue stem cell. PLoS Biol. 3, 1596–1606 (2005)

    Article  Google Scholar 

  13. Pollard, S. M., Conti, L., Sun, Y., Goffredo, D. & Smith, A. Adherent neural stem (NS) cells from foetal and adult forebrain. Cerebral Cortex (in the press)

  14. Niwa, H. et al. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell 123, 917–929 (2005)

    Article  CAS  PubMed  Google Scholar 

  15. Smith, A. The battlefield of pluripotency. Cell 123, 757–760 (2005)

    Article  CAS  PubMed  Google Scholar 

  16. Bao, S. et al. Initiation of epigenetic reprogramming of the X chromosome in somatic nuclei transplanted to a mouse oocyte. EMBO Rep. 6, 748–754 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Chambers, I. & Smith, A. Self-renewal of teratocarcinoma and embryonic stem cells. Oncogene 23, 7150–7160 (2004)

    Article  CAS  PubMed  Google Scholar 

  18. Yeom, Y. I. et al. Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells. Development 122, 881–894 (1996)

    CAS  PubMed  Google Scholar 

  19. Pratt, T., Sharp, L., Nichols, J., Price, D. J. & Mason, J. O. Embryonic stem cells and transgenic mice ubiquitously expressing a tau-tagged green fluorescent protein. Dev. Biol. 228, 19–28 (2000)

    Article  CAS  PubMed  Google Scholar 

  20. Yates, A. & Chambers, I. The homeodomain protein Nanog and pluripotency in mouse embryonic stem cells. Biochem. Soc. Trans. 33, 1518–1521 (2005)

    Article  CAS  PubMed  Google Scholar 

  21. Silva, J. et al. Establishment of histone h3 methylation on the inactive X chromosome requires transient recruitment of Eed–Enx1 polycomb group complexes. Dev. Cell 4, 481–495 (2003)

    Article  CAS  PubMed  Google Scholar 

  22. de Napoles, M. et al. Polycomb group proteins Ring1A/B link ubiquitylation of histone H2A to heritable gene silencing and X inactivation. Dev. Cell 7, 663–676 (2004)

    Article  CAS  PubMed  Google Scholar 

  23. Sheardown, S. A. et al. Stabilization of Xist RNA mediates initiation of X chromosome inactivation. Cell 91, 99–107 (1997)

    Article  CAS  PubMed  Google Scholar 

  24. Mountford, P. et al. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. Proc. Natl Acad. Sci. USA 91, 4303–4307 (1994)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  25. Mountford, P., Nichols, J., Zevnik, B., O'Brien, C. & Smith, A. Maintenance of pluripotential embryonic stem cells by stem cell selection. Reprod. Fertil. Dev. 10, 527–533 (1998)

    Article  CAS  PubMed  Google Scholar 

  26. Do, J. T. & Scholer, H. R. Nuclei of embryonic stem cells reprogram somatic cells. Stem Cells 22, 941–949 (2004)

    Article  CAS  PubMed  Google Scholar 

  27. Blelloch, R. et al. Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus. Stem Cells doi:10.1634/stemcells.2006-0050 (18 May 2006)

  28. Avilion, A. A. et al. Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev. 17, 126–140 (2003)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank J. Vrana for assistance with FACS and Y. Costa for critical reading of the manuscript. This research was supported by the Biotechnological and Biological Sciences Research Council and the Medical Research Council of the United Kingdom, the Wellcome Trust, and by an EMBO Long-term Fellowship (J.S.). Author Contributions J.S. designed and executed experiments, analysed data, and drafted the paper; I.C. and S.P. generated reagents and contributed to experimental design; and A.S. designed experiments and wrote the paper.

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Correspondence to Austin Smith.

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The University of Edinburgh has filed a patent application related to this work and has licensed this patent to Stem Cell Sciences, PLC. A.S. is a scientific consultant to Stem Cell Sciences, PLC.

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This file contains Supplementary Methods, Supplementary Figure Legends, Supplementary Figures 1–8 and additional references. (PDF 3715 kb)

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Silva, J., Chambers, I., Pollard, S. et al. Nanog promotes transfer of pluripotency after cell fusion. Nature 441, 997–1001 (2006). https://doi.org/10.1038/nature04914

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