Primordial germ cells (PGCs) are the precursors of sperm and eggs1. In most animals, segregation of the germ line from the somatic lineages is one of the earliest events in development2; in avian embryos, PGCs are first identified in an extra-embryonic region, the germinal crescent, after approximately 18 h of incubation. After 50–55 h of development, PGCs migrate to the gonad and subsequently produce functional sperm and oocytes3,4. So far, cultures of PGCs that remain restricted to the germ line have not been reported in any species5,6. Here we show that chicken PGCs can be isolated, cultured and genetically modified while maintaining their commitment to the germ line. Furthermore, we show that chicken PGCs can be induced in vitro to differentiate into embryonic germ cells that contribute to somatic tissues. Retention of the commitment of PGCs to the germ line after extended periods in culture and after genetic modification combined with their capacity to acquire somatic competence in vitro provides a new model for developmental biology. The utility of the model is enhanced by the accessibility of the avian embryo, which facilitates access to the earliest stages of development and supplies a facile route for the reintroduction of PGCs into the embryonic vasculature. In addition, these attributes create new opportunities to manipulate the genome of chickens for agricultural and pharmaceutical applications.

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  1. 1

    Nieuwkoop, P. & Sutasurya, L. Primordial Germ Cells in the Chordates (Cambridge Univ. Press, New York, 1979)

  2. 2

    Wylie, C. Germ cells. Cell 96, 165–174 (1999)

  3. 3

    Fujimoto, T., Ninomiya, T. & Ukeshima, A. Observations of the primordial germ cells in blood samples from the chick embryo. Dev. Biol. 49, 278–282 (1976)

  4. 4

    Swift, C. H. Origin and early history of the primordial germ-cells in the chick. Am. J. Anat. 15, 483–516 (1914)

  5. 5

    McLaren, A. The quest for immortality. Nature 359, 482–483 (1992)

  6. 6

    McLaren, A. Primordial germ cells in the mouse. Dev. Biol. 262, 1–15 (2003)

  7. 7

    Hamburger, V. & Hamilton, H. A series of normal stages in the development of the chick embryo. J. Morphol. 88, 49–92 (1951)

  8. 8

    Kodama, H. et al. Nucleotide sequences and unusual electrophoretic behavior of the W chromosome-specific repeating DNA units of the domestic fowl, Gallus gallus domesticus. Chromosoma 96, 18–25 (1987)

  9. 9

    Tsunekawa, N., Naito, M., Sakai, Y., Nishida, T. & Noce, T. Isolation of chicken vasa homolog gene and tracing the origin of primordial germ cells. Development 127, 2741–2750 (2000)

  10. 10

    Halfter, W. et al. An ovomucin-like protein on the surface of migrating primordial germ cells of the chick and rat. Development 122, 915–923 (1996)

  11. 11

    van de Lavoir, M.-C. et al. High grade transgenic somatic chimeras from chicken embryonic stem cells. Mech. Dev. 123, 31–41 (2006)

  12. 12

    Baba, T. W., Giroir, B. P. & Humphries, E. H. Cell lines derived from avian lymphomas exhibit two distinct phenotypes. Virology 144, 139–151 (1985)

  13. 13

    Naito, M. et al. Differentiation of donor primordial germ cells into functional gametes in the gonads of mixed-sex germline chimaeric chickens produced by transfer of primordial germ cells isolated from embryonic blood. J. Reprod. Fertil. 117, 291–298 (1999)

  14. 14

    Burgess-Beusse, B. et al. The insulation of genes from external enhancers and silencing chromatin. Proc. Natl Acad. Sci. USA 99 (suppl. 4), 16433–16437 (2002)

  15. 15

    Eyal-Giladi, H. & Kochav, S. From cleavage to primitive streak formation: a complimentary normal table and a new look at the first stages of the development of the chick. Dev. Biol. 49, 321–337 (1973)

  16. 16

    Stern, C. D. The chick: A great model system becomes even greater. Dev. Cell 8, 9–17 (2005)

  17. 17

    Zhu, L. et al. Production of human monoclonal antibody in eggs of chimeric chickens. Nature Biotechnol. 23, 1159–1169 (2005)

  18. 18

    Petitte, J. N. in Handbook of Stem Cells (eds Lanza, R. et al.) 471–477 (Elsevier Academic Press, Burlington, Massachusetts, 2004)

  19. 19

    Kim, N. et al. Specific association of human telomerase activity with immortal cells and cancer. Science 266, 2011–2014 (1994)

  20. 20

    Streit, A., Berliner, A. J., Papanayotou, C., Sirulnik, A. & Stern, C. D. Initiation of neural induction by FGF signalling before gastrulation. Nature 406, 74–78 (2000)

  21. 21

    Niwa, H., Yamamura, K. & Miyazaki, J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199 (1991)

  22. 22

    Rowlett, K. & Simkiss, K. Explanted embryo culture: in vitro and in ovo techniques for domestic fowl. Br. Poult. Sci. 28, 91–101 (1987)

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We thank C. Gitter for technical assistance with the karyotype; A. Pradas-Monne for help in the laboratory; W. Halfter for providing the 1B3 antibody; Leica for the provision of optical equipment to photograph the GFP-positive embryo; and J.-M. Buerstedde for supplying β-actin-neo and β-actin-puro. This work was supported by the Small Business Innovation Research Programs of the USDA and the NIH to Origen Therapeutics and a USDA grant to M.E.D. Author Contributions M.C.L. developed the cell culture system with the assistance of J.H.D., P.A.L. and R.B.; C.M.-L. and J.H.D. performed the embryological manipulations; P.A.L. executed the molecular biology in collaboration with B.S.H. and L.T.H.; A.K. provided animal care; T.M.G., S.E.S. and M.E.D. conducted the telomerase assay and karyotyping; M.C.L. and R.J.E. coordinated the contributions of authors and wrote the paper. All authors discussed the results and commented on the manuscript.

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  1. Origen Therapeutics, Burlingame, 1450 Rollins Road, California, 94010, USA

    • Marie-Cecile van de Lavoir
    • , Jennifer H. Diamond
    • , Philip A. Leighton
    • , Christine Mather-Love
    • , Babette S. Heyer
    • , Renee Bradshaw
    • , Allyn Kerchner
    • , Lisa T. Hooi
    •  & Robert J. Etches
  2. Department of Animal Science, University of California, 1 Shields Avenue, Meyer Hall, Davis, 2131D, California, 95616, Davis, USA

    • Terri M. Gessaro
    • , Susan E. Swanberg
    •  & Mary E. Delany


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Competing interests

Some of the authors are employees at Origen Therapeutics.

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Correspondence to Robert J. Etches.

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