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Conversion of adult mouse unipotent germline stem cells into pluripotent stem cells

Nature Protocols volume 5, pages 921–928 (2010)Cite this article

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Abstract

Germline stem cells (GSCs), often called spermatogonial stem cells, are unipotent stem cells that can give rise only to gametes. Under defined culture conditions, unipotent GSCs can be converted into pluripotent stem cells, termed as germline-derived pluripotent stem (gPS) cells. gPS cells can be differentiated into cells forming all three germ layers and germ cells. In this study, we describe a simple and robust protocol for the derivation of GSCs from adult mouse testis and the rapid and reproducible conversion of GSCs into gPS cells. Under our defined culture conditions, GSCs can be converted into gPS cells in ∼1 month. The initial number of plated GSCs and the culture time are two conditions that are critical for the successful conversion of GSCs into gPS cells. gPS cells are similar to embryonic stem cells, as judged by molecular and cellular properties and its development potential. Thus, generation of gPS cells holds potential for tissue regenerative medicine.

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Figure 1
Figure 2: Establishment of GSC lines from either an Oct4-GFP transgenic or a wild-type mouse.
Figure 3: Characterization of established GSCs from an Oct4-GFP transgenic mouse.
Figure 4: Conversion of GSCs into gPS cells.
Figure 5: Cellular and molecular characterization of established Oct4-GFP gPS cells.

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References

  1. Tegelenbosch, R.A. & de Rooij, D.G. A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H/101 F1 hybrid mouse. Mutat. Res. 290, 193–200 (1993).

    Article  CAS  PubMed  Google Scholar 

  2. Kanatsu-Shinohara, M. et al. Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol. Reprod. 69, 612–616 (2003).

    Article  CAS  PubMed  Google Scholar 

  3. Kubota, H., Avarbock, M.R. & Brinster, R.L. Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proc. Natl. Acad. Sci. USA 101, 16489–16494 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ko, K. et al. Induction of pluripotency in adult unipotent germline stem cells. Cell Stem Cell 5, 87–96 (2009).

    Article  CAS  PubMed  Google Scholar 

  5. Ogawa, T. et al. Derivation and morphological characterization of mouse spermatogonial stem cell lines. Arch. Histol. Cytol. 67, 297–306 (2004).

    Article  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  7. Seandel, M. et al. Generation of functional multipotent adult stem cells from GPR125+ germline progenitors. Nature 449, 346–350 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Izadyar, F. et al. Generation of multipotent cell lines from a distinct population of male germ line stem cells. Reproduction 135, 771–784 (2008).

    Article  CAS  PubMed  Google Scholar 

  9. Kanatsu-Shinohara, M. et al. Pluripotency of a single spermatogonial stem cell in mice. Biol. Reprod. 78, 681–687 (2008).

    Article  CAS  PubMed  Google Scholar 

  10. Guan, K. et al. Pluripotency of spermatogonial stem cells from adult mouse testis. Nature 440, 1199–1203 (2006).

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Martin Zenke, from RWTH Aachen, for conducting microarray experiments and Jeanine Müller-Keuker for preparing the illustrations. This work has been supported in part by the Deutsche Forschungsgemeinschaft DFG grant SCHO 340/4-1 and by the German Federal Ministry of Education and Research BMBF grant S01GN 0811.

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Authors and Affiliations

  1. Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, NRW, Germany

    Kinarm Ko, Marcos J Araúzo-Bravo, Julee Kim, Martin Stehling & Hans R Schöler

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  1. Kinarm Ko
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  2. Marcos J Araúzo-Bravo
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  3. Julee Kim
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  4. Martin Stehling
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  5. Hans R Schöler
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Contributions

K.K. designed the protocol, generated and characterized the GSC and gPS cells and wrote the paper. M.J.A.-B., J.K. and M.S. characterized the GSC and gPS cells. H.R.S. designed the protocol and wrote the paper.

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Correspondence to Hans R Schöler.

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The authors declare no competing financial interests.

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Ko, K., Araúzo-Bravo, M., Kim, J. et al. Conversion of adult mouse unipotent germline stem cells into pluripotent stem cells. Nat Protoc 5, 921–928 (2010). https://doi.org/10.1038/nprot.2010.44

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