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Mouse differentiating spermatogonia can generate germinal stem cells in vivo

Nature Cell Biology volume 11, pages 190–196 (2009)Cite this article

Abstract

In adults, stem cells are responsible for the maintenance of many actively renewing tissues, such as haematopoietic, skin, gut and germinal tissues. These stem cells can self-renew or be committed to becoming progenitors. Stem-cell commitment is thought to be irreversible but in male and female Drosophila melanogaster, it was shown recently that differentiating germ cells can revert to functional stem cells that can restore germinal lineage1,2. Whether progenitors are also able to generate stem cells in mammals remains unknown. Here we show that purified mouse spermatogonial progenitors committed to differentiation can generate functional germinal stem cells that can repopulate germ-cell-depleted testes when transplanted into adult mice. We found that GDNF, a key regulator of the stem-cell niche, and FGF2 are able to reprogram in vitro spermatogonial progenitors for reverse differentiation. This study supports the emerging concept that the stem-cell identity is not restricted in adults to a definite pool of cells that self-renew, but that stemness could be acquired by differentiating progenitors after tissue injury and throughout life.

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Figure 1: Adult c-kit-positive spermatogonial progenitors selected by MACS have the potential to regenerate spermatogenesis after transplantation in germ-cell depleted testis.
Figure 2: Flow cytometric analysis of c-kit and stem-cell activity in SPα-6+c-kit−/low subpopulation of wild-type adult testes cells.
Figure 3: Adult SPα6+c-kit+ cells differentiating spermatogonia have the potential to regenerate spermatogenesis after transplantation.
Figure 4: Differentiating c-kit-positive spermatogonia de-differentiate and act as functional GSCs after transplantation.
Figure 5: Chemoattraction of c-kit positive spermatogonia by KitL in vitro, and generation of germ-cell clusters with long-term clonogenic activity from c-kit positive spermatogonia in culture in presence of GDNF and FGF2.

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Acknowledgements

We thank P.H. Romeo for critical review of the manuscript and helpful comments. We thank M. Okabe for the generous gift of the EGFP transgenic mice; S. Leblay and V. Neuville for their technical assistance in the animal facilities, and T. Andrieu for assistance with Aria, LSR and LSRII. This work was supported in part by a grant from Electricité De France.

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Author notes

  1. Vilma Barroca and Bruno Lassalle: These authors contributed equally to this work

Authors and Affiliations

  1. Laboratoire Gamétogenèse, Apoptose et Génotoxicité, INSERM U566, Institut de Radiobiologie Cellulaire et Moléculaire, Direction des Sciences du Vivant, CEA, Fontenay aux Roses, 92265, France

    Vilma Barroca, Bruno Lassalle, Mathieu Coureuil, Florence Le Page, Jacques Testart, Isabelle Allemand, Lydia Riou & Pierre Fouchet

  2. UMR 6218, CNRS, Orléans, 45071, France

    Jean Paul Louis

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Contributions

V.B. and B.L. contributed equally to the experimental work, with help from M.C., L.R.and P.F.; V.B., B.L., L.R. and P.F. conceived and designed the experiments; J.P.L. provided valuable material and initiated the breeding of the EGFP mice; F.L.P., J.T., I.A., B.L. and L.R. revised the manuscript. All authors participated in data analysis. P.F. coordinated the study; P.F. and V.B. wrote the paper.

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Correspondence to Pierre Fouchet.

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Barroca, V., Lassalle, B., Coureuil, M. et al. Mouse differentiating spermatogonia can generate germinal stem cells in vivo. Nat Cell Biol 11, 190–196 (2009). https://doi.org/10.1038/ncb1826

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