Letter | Published:

Pluripotency of spermatogonial stem cells from adult mouse testis

Nature volume 440, pages 11991203 (27 April 2006) | Download Citation

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Abstract

Embryonic germ cells as well as germline stem cells from neonatal mouse testis are pluripotent and have differentiation potential similar to embryonic stem cells1,2, suggesting that the germline lineage may retain the ability to generate pluripotent cells. However, until now there has been no evidence for the pluripotency and plasticity of adult spermatogonial stem cells (SSCs), which are responsible for maintaining spermatogenesis throughout life in the male3. Here we show the isolation of SSCs from adult mouse testis using genetic selection, with a success rate of 27%. These isolated SSCs respond to culture conditions and acquire embryonic stem cell properties. We name these cells multipotent adult germline stem cells (maGSCs). They are able to spontaneously differentiate into derivatives of the three embryonic germ layers in vitro and generate teratomas in immunodeficient mice. When injected into an early blastocyst, SSCs contribute to the development of various organs and show germline transmission. Thus, the capacity to form multipotent cells persists in adult mouse testis. Establishment of human maGSCs from testicular biopsies may allow individual cell-based therapy without the ethical and immunological problems associated with human embryonic stem cells. Furthermore, these cells may provide new opportunities to study genetic diseases in various cell lineages.

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References

  1. 1.

    , & Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture. Cell 70, 841–847 (1992)

  2. 2.

    et al. Generation of pluripotent stem cells from neonatal mouse testis. Cell 119, 1001–1012 (2004)

  3. 3.

    , & Stem cells find their niche. Nature 414, 98–104 (2001)

  4. 4.

    et al. Characterization of a premeiotic germ cell-specific cytoplasmic protein encoded by Stra8, a novel retinoic acid-responsive gene. J. Cell Biol. 135, 469–477 (1996)

  5. 5.

    et al. Murine spermatogonial stem cells: targeted transgene expression and purification in an active state. EMBO Rep. 3, 753–759 (2002)

  6. 6.

    et al. Stem cell based therapeutical approach of male infertility by teratocarcinoma derived germ cells. Hum. Mol. Genet. 13, 1451–1460 (2004)

  7. 7.

    & A description of spontaneous congenital testicular teratomas in strain 129 mice. J. Natl. Cancer Inst. 18, 719–747 (1957)

  8. 8.

    et al. Effects of genetic background on tumorigenesis in p53-deficient mice. Mol. Carcinog. 14, 16–22 (1995)

  9. 9.

    & Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1). Proc. Natl Acad. Sci. USA 75, 5565–5569 (1978)

  10. 10.

    et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95, 379–391 (1998)

  11. 11.

    et al. Immunohistochemical localization of murine stage-specific embryonic antigens in human testicular germ cell tumors. Am. J. Pathol. 108, 225–230 (1982)

  12. 12.

    , , & Role of c-kit in mammalian spermatogenesis. J. Endocrinol. Invest. 23, 609–615 (2000)

  13. 13.

    , & Culture conditions and single growth factors affect fate determination of mouse spermatogonial stem cells. Biol. Reprod. 71, 722–731 (2004)

  14. 14.

    & In vitro differentiation of embryonic stem cells: immunophenotypic analysis of cultured embryoid bodies. J. Cell. Physiol. 171, 104–115 (1997)

  15. 15.

    , , , & Maintenance of mouse male germ line stem cells in vitro. Biol. Reprod. 68, 2207–2214 (2003)

  16. 16.

    , & Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proc. Natl Acad. Sci. USA 101, 16489–16494 (2004)

  17. 17.

    , & Gene expression profiling of mouse teratocarcinomas uncovers epigenetic changes associated with the transformation of mouse embryonic stem cells. Neoplasia 6, 490–502 (2004)

  18. 18.

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

  19. 19.

    et al. UTF1, a novel transcriptional coactivator expressed in pluripotent embryonic stem cells and extra-embryonic cells. EMBO J. 17, 2019–2032 (1998)

  20. 20.

    et al. Gene expression profiling of embryo-derived stem cells reveals candidate genes associated with pluripotency and lineage specificity. Genome Res. 12, 1921–1928 (2002)

  21. 21.

    , , & Rex-1, a gene encoding a transcription factor expressed in the early embryo, is regulated via Oct-3/4 and Oct-6 binding to an octamer site and a novel protein, Rox-1, binding to an adjacent site. Mol. Cell. Biol. 18, 1866–1878 (1998)

  22. 22.

    , & Embryonic stem cell differentiation models: cardiogenesis, myogenesis, neurogenesis, epithelial and vascular smooth muscle cell differentiation in vitro. Cytotechnology 30, 211–226 (1999)

  23. 23.

    , & The role of the brachyury gene in heart development and left-right specification in the mouse. Mech. Dev. 79, 29–37 (1998)

  24. 24.

    et al. Postnatal isl1+ cardioblasts enter fully differentiated cardiomyocyte lineages. Nature 433, 647–653 (2005)

  25. 25.

    , & Myogenesis in the mouse embryo: differential onset of expression of myogenic proteins and the involvement of titin in myofibril assembly. J. Cell Biol. 109, 517–527 (1989)

  26. 26.

    et al. HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J. Clin. Invest. 108, 391–397 (2001)

  27. 27.

    et al. Improved generation of germline-competent embryonic stem cell lines from inbred mouse strains. Stem Cells 21, 90–97 (2003)

  28. 28.

    , & CD9 is a surface marker on mouse and rat male germline stem cells. Biol. Reprod. 70, 70–75 (2004)

  29. 29.

    et al. Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287, 1489–1493 (2000)

  30. 30.

    , , , & Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810–813 (1997)

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Acknowledgements

We thank A. Cierpka, D. Meyer, S. Wolf, B. Sadowski, I. Schwandt, C. Müller and S. Burkhardt for technical assistance. We thank M. Schindler, H. Riedesel and S. Wolf for their assistance in the generation of transgenic mice, G. Wulf for help with FACS analysis, and B. Hemmerlein for the analysis of teratomas. This work was supported by grants from the Georg-August-University of Göttingen (Forschungsförderungsprogramm Stammzellen) to K.G. and K.N., and a DFG grant (Emmy-Noether Program) to L.S.M. Author Contributions K.G., G.H., K.N. and W.E. conceived and designed the experiments and performed the data analysis and controls. K.G., K.N., L.S.M., S.W., R.D., J.H.L., J.N., F.W. and M.L. performed the experiments, and K.G., G.H., K.N. and W.E. wrote the paper.

Author information

Author notes

    • Kaomei Guan
    •  & Karim Nayernia

    These authors contributed equally to this work

Affiliations

  1. Department of Cardiology and Pneumology, Heart Center, Georg-August-University of Göttingen, Robert-Koch-Str. 40, 37075 Göttingen, Germany

    • Kaomei Guan
    • , Lars S. Maier
    • , Stefan Wagner
    • , Frieder Wolf
    •  & Gerd Hasenfuss
  2. Institute of Human Genetics and

    • Karim Nayernia
    • , Jae Ho Lee
    • , Jessica Nolte
    • , Manyu Li
    •  & Wolfgang Engel
  3. Department of Cellular and Molecular Immunology, Georg-August-University of Göttingen, Heinrich-Dücker-Weg 12, 37073 Göttingen, Germany

    • Ralf Dressel

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Competing interests

Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Corresponding authors

Correspondence to Wolfgang Engel or Gerd Hasenfuss.

Supplementary information

Word documents

  1. 1.

    Supplementary Methods

    Full description of methods and analysis used in this study.

  2. 2.

    Supplementary Tables

    This file contains Supplementary Tables 1-4. Analysis of DNA microsatellite markers in the established cell lines as compared to the other cells cultured in the same facility.

  3. 3.

    Supplementary Video Legends

    This file contains text to accompany the above Supplementary Video.

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    Supplementary Figures and Figure Legends

    This file contains Supplementary Figures 1-6.

Videos

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    Supplementary Video 1

    Spontaneously and rhythmically beating cells in culture.

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DOI

https://doi.org/10.1038/nature04697

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