Generation of functional multipotent adult stem cells from GPR125+ germline progenitors (original) (raw)

Nature volume 449, pages 346–350 (2007)Cite this article

Abstract

Adult mammalian testis is a source of pluripotent stem cells1. However, the lack of specific surface markers has hampered identification and tracking of the unrecognized subset of germ cells that gives rise to multipotent cells2. Although embryonic-like cells can be derived from adult testis cultures after only several weeks in vitro1, it is not known whether adult self-renewing spermatogonia in long-term culture can generate such stem cells as well. Here, we show that highly proliferative adult spermatogonial progenitor cells (SPCs) can be efficiently obtained by cultivation on mitotically inactivated testicular feeders containing CD34+ stromal cells. SPCs exhibit testicular repopulating activity in vivo and maintain the ability in long-term culture to give rise to multipotent adult spermatogonial-derived stem cells (MASCs). Furthermore, both SPCs and MASCs express GPR125, an orphan adhesion-type G-protein-coupled receptor. In knock-in mice bearing a GPR125–β-galactosidase (LacZ) fusion protein under control of the native Gpr125 promoter (GPR125–LacZ), expression in the testis was detected exclusively in spermatogonia and not in differentiated germ cells. Primary GPR125–LacZ SPC lines retained GPR125 expression, underwent clonal expansion, maintained the phenotype of germline stem cells, and reconstituted spermatogenesis in busulphan-treated mice. Long-term cultures of GPR125+ SPCs (GSPCs) also converted into GPR125+ MASC colonies. GPR125+ MASCs generated derivatives of the three germ layers and contributed to chimaeric embryos, with concomitant downregulation of GPR125 during differentiation into GPR125- cells. MASCs also differentiated into contractile cardiac tissue in vitro and formed functional blood vessels in vivo. Molecular bookmarking by GPR125 in the adult mouse and, ultimately, in the human testis could enrich for a population of SPCs for derivation of GPR125+ MASCs, which may be employed for genetic manipulation, tissue regeneration and revascularization of ischaemic organs.

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Acknowledgements

This work was supported by the Howard Hughes Medical Institute, Ansary Stem Cell Center for Regenerative Medicine and Memorial Sloan Kettering Cancer Center T32 grant (M.S.), an AACR–Genentech BioOncology Fellowship for Cancer Research on Angiogenesis (M.S.), the Heed Foundation (S.C.), the International Retinal Research Foundation (S.C.) and National Heart, Lung and Blood Institute grants (S.R.). We thank M. Hardy, P. Schlegel, Marc Goldstein, A. Brivanlou and S. Noggle for critical input. We are grateful to G. Enders for providing anti-GCNA antibody. We thank D. S. Johnston, G. Linkov and G. Zlotchenko for technical assistance.

Author information

Authors and Affiliations

  1. Department of Genetic Medicine, Howard Hughes Medical Institute, and,
    Marco Seandel, Daylon James, Sergey V. Shmelkov, Ilaria Falciatori, Jiyeon Kim, Sai Chavala, Fan Zhang & Shahin Rafii
  2. Department of Urology, Weill Cornell Medical College, New York 10065, USA,
    Douglas S. Scherr
  3. Division of Medical Oncology, Department of Medicine, and,
    Marco Seandel
  4. Department of Pathology, Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York 10065, USA,
    Robin M. Hobbs & Pier Paolo Pandolfi
  5. Regeneron Pharmaceuticals, Tarrytown, New York 10591, USA,
    Richard Torres, Nicholas W. Gale, George D. Yancopoulos, Andrew Murphy & David M. Valenzuela
  6. Cancer Genetics Program, Beth Israel Deaconess Cancer Center, Harvard Medical School, Boston, Massachusetts 02115, USA,
    Robin M. Hobbs & Pier Paolo Pandolfi

Authors

  1. Marco Seandel
  2. Daylon James
  3. Sergey V. Shmelkov
  4. Ilaria Falciatori
  5. Jiyeon Kim
  6. Sai Chavala
  7. Douglas S. Scherr
  8. Fan Zhang
  9. Richard Torres
  10. Nicholas W. Gale
  11. George D. Yancopoulos
  12. Andrew Murphy
  13. David M. Valenzuela
  14. Robin M. Hobbs
  15. Pier Paolo Pandolfi
  16. Shahin Rafii

Corresponding author

Correspondence toShahin Rafii.

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Competing interests

S.R., M.S., S.V.S. and S.C. have filed a provisional patent application related to the use of GPR125.

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Supplementary Information

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Seandel, M., James, D., Shmelkov, S. et al. Generation of functional multipotent adult stem cells from GPR125+ germline progenitors.Nature 449, 346–350 (2007). https://doi.org/10.1038/nature06129

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Editorial Summary

Stem cells make their mark

Adult stem cells are an attractive alternative to embryonic stem cells for therapeutic use. As yet there is no standard method for obtaining such cells from adults and priming them to form different tissues, but a new system that generates large numbers of stem cells from the adult testicle shows promise. It makes use of a novel marker, an orphan receptor known as GPR125, found on the surface of spermatogonial stem cells. The use of specialized feeder cells to support stem cell growth allows stem cells once destined for spermatogenesis to become multipotent. This work also provides clues as to the minimal requirements for multipotency in adult cells.