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Germline transmission of genetically modified primordial germ cells

Nature volume 441, pages 766–769 (2006)Cite this article

Abstract

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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Figure 1: Characteristics of PGC cell lines in vitro.
Figure 2: FACS analysis of DT40 cells, ES cells, EG cells and PGCs, stained with antibodies against CVH and OLP.
Figure 3: β-Actin-EGFP is incorporated into the PGC genome and expressed in offspring.

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References

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

    Google Scholar 

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

    Article  CAS  Google Scholar 

  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)

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  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)

    Article  CAS  Google Scholar 

  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)

    CAS  PubMed  Google Scholar 

  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)

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  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)

    Article  CAS  Google Scholar 

  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)

    Article  CAS  Google Scholar 

  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)

    Article  ADS  CAS  Google Scholar 

  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)

    Article  Google Scholar 

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

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Book  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  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)

    Article  ADS  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

Download references

Acknowledgements

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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Authors and Affiliations

  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

Authors
  1. Marie-Cecile van de Lavoir
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  2. Jennifer H. Diamond
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Correspondence to Robert J. Etches.

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Some of the authors are employees at Origen Therapeutics.

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Supplementary Notes

This file contains Supplementary Figure 1 and Supplementary Table 1. (PDF 4764 kb)

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van de Lavoir, MC., Diamond, J., Leighton, P. et al. Germline transmission of genetically modified primordial germ cells. Nature 441, 766–769 (2006). https://doi.org/10.1038/nature04831

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