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Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei

Nature volume 394, pages 369–374 (1998)Cite this article

Abstract

Until recently, fertilization was the only way to produce viable mammalian offspring, a process implicitly involving male and female gametes. However, techniques involving fusion of embryonic or fetal somatic cells with enucleated oocytes have become steadily more successful in generating cloned young1,2,3. Dolly the sheep4 was produced by electrofusion of sheep mammary-derived cells with enucleated sheep oocytes. Here we investigate the factors governing embryonic development by introducing nuclei from somatic cells (Sertoli, neuronal and cumulus cells) taken from adult mice into enucleated mouse oocytes. We found that some enucleated oocytes receiving Sertoli or neuronal nuclei developed in vitro and implanted following transfer, but none developed beyond 8.5 days post coitum; however, a high percentage of enucleated oocytes receiving cumulus nuclei developed in vitro. Once transferred, many of these embryos implanted and, although most were subsequently resorbed, a significant proportion (2 to 2.8%) developed to term. These experiments show that for mammals, nuclei from terminally differentiated, adult somatic cells of known phenotype introduced into enucleated oocytes are capable of supporting full development.

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Figure 1: In vitro development of enucleated oocytes following injection of cumulus cell nuclei.
Figure 2: Cloned mice.
Figure 3: Development following uterine transfer of embryos produced after injection of Sertoli cell nuclei into enucleated oocytes.
Figure 4: DNA typing of donors and offspring in series C corroborates the genetic identify of the cloned offspring to cumulus cell donors, and non-identity to oocyte donors and host foster females.
Figure 5

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References

  1. Campbell, K. H. S., Loi, P., Otaegui, P. J. & Wilmut, I. Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev. Reprod. 1, 40–45 (1996).

    Article  CAS  Google Scholar 

  2. Kono, T. Nuclear transfer and reprogramming. Rev. Reprod. 2, 74–80 (1997).

    Article  CAS  Google Scholar 

  3. Campbell, K. H. S., McWhir, J., Ritchie, W. A. & Wilmut, I. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–66 (1996).

    Article  ADS  CAS  Google Scholar 

  4. Wilmut, I., Schnieke, A. E., McWhir, J., Kind, A. J. & Campbell, K. H. S. Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810–813 (1997).

    Article  ADS  CAS  Google Scholar 

  5. Schuetz, A. W., Whittingham, D. G. & Snowden, R. Alterations in the cell cycle of mouse cumulus granulosa cells during expansion and mucification in vivo and in vitro. Reprod. Fertil. Dev. 8, 935–943 (1996).

    Article  CAS  Google Scholar 

  6. Wassarman, P. The biology and chemistry of fertilization. Science 235, 553–560 (1987).

    Article  ADS  CAS  Google Scholar 

  7. Epstein, C. J. The Consequences of Chromosome Imbalance (Cambridge University Press, 1986).

    Book  Google Scholar 

  8. Yanagida, K., Yanagimachi, R., Perreault, S. D. & Kleinfeld, R. G. Thermostabiity of sperm nuclei assessed by microinjection into hamster oocytes. Biol. Reprod. 44, 440–447 (1991).

    Article  CAS  Google Scholar 

  9. McGrath, J. & Solter, D. Inability of mouse blastmere nuclei transferred to enucleated zygotes to support development in vitro. Science 226, 1317–1319 (1984).

    Article  ADS  CAS  Google Scholar 

  10. Willadsen, S. M. Nuclear transplantation in sheep embryos. Nature 320, 63–65 (1986).

    Article  ADS  CAS  Google Scholar 

  11. Collas, P. & Barnes, F. L. Nuclear transplantation by microinjection of inner cell mass and granulosa cell nuclei. Mol. Reprod. Dev. 38, 264–267 (1994).

    Article  CAS  Google Scholar 

  12. Czolowska, R., Modlinski, J. A. & Tarkowski, A. K. Behavior of thymocyte nuclei in non-activated and activated mouse oocytes. J. Cell Sci. 69, 19–34 (1984).

    CAS  PubMed  Google Scholar 

  13. Tsunoda, T. & Kato, Y. Nuclear transplantation of embryonic stem cells in mice. J. Reprod. Fertil. 98, 537–540 (1993).

    Article  CAS  Google Scholar 

  14. Tsunoda, T., Tokunaga, T., Imai, I. & Uchida, T. Nuclear transplantation of male primordial germ cells in the mouse. Development 107, 407–411 (1989).

    CAS  PubMed  Google Scholar 

  15. Kono, T., Ogawa, M. & Nakahara, T. Thymocyte transfer to enucleated oocytes in the mouse. J. Reprod. Dev. 39, 301–307 (1993).

    Article  Google Scholar 

  16. Kimura, Y. & Yanagimachi, R. Intracytoplasmic sperm injection in the mouse. Biol. Reprod. 52, 709–720 (1995).

    Article  CAS  Google Scholar 

  17. Kimura, Y. & Yanagimachi, R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring. Development 121, 2397–2405 (1995).

    CAS  PubMed  Google Scholar 

  18. Eppig, J., Wigglesworth, K., Pendola, F. & Hirao, Y. Murine oocytes suppress expression of luteinizing hormone receptor messenger ribonucleic acid by granulosa cells. Biol. Reprod. 56, 976–984 (1997).

    Article  CAS  Google Scholar 

  19. Chatot, C. L., Lewis, J. L., Torres, I. & Ziomek, C. A. Development of 1-cell embryos from different strains of mice in CZB medium. Biol. Reprod. 42, 432–440 (1990).

    Article  CAS  Google Scholar 

  20. Erickson, R. P., Zwigman, T. & Ao, A. Gene expression, X-inactivation, and methylation during spermatogenesis: the case of Zfa, Zfx and Zfy in mice. Mol. Reprod. Dev. 35, 114–120 (1993).

    Article  CAS  Google Scholar 

  21. 1. Kuretake, S., Kimura, Y., Hoshi, K. & Yanagimachi, R. Fertilization and development of mouse oocytes injected with isolated sperm heads. Biol. Reprod. 55, 789–795 (1996).

    Article  CAS  Google Scholar 

  22. Kono, T., Sotomaru, Y., Sato, Y. & Nakahara, T. Development of androgenetic mouse embryos produced by in vitro fertilization of enucleated oocytes. Mol. Reprod. Dev. 34, 43–46 (1993).

    Article  CAS  Google Scholar 

  23. Bos-Mikich, A., Whittingham, D. G. & Kones, K. T. Meiotic and Mitotic Ca2+ oscillations affect cell composition in resulting blastocysts. Dev. Biol. 182, 172–179 (1997).

    Article  CAS  Google Scholar 

  24. Dietrich, W. et al. Agenetic map of the mouse suitable for typing intraspecific crosses. Genetics 131, 423–447 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Taylor, B. A. & Rowe, L. Amouse linkage testing stock possessing multiple copies of the endogenous ecotropic murine leukemia virus genome. Genomics 5, 221–232 (1989).

    Article  CAS  Google Scholar 

  26. Johnson, K. R., Cook, S. A. & Davisson, M. T. Chromosomal localization of the murine gene and two related sequences encoding high-mobility-group I and Y proteins. Genomics 12, 503–509 (1992).

    Article  CAS  Google Scholar 

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Acknowledgements

This study was supported in part by grants from the National Institutes of Health, ProBio America Inc., and fellowships from the Japanese Society for the Promotion of Science (T.W.) and European Molecular Biology Organization (A.C.F.P.). We thank H. Tateno for chromosome analysis, Y.Nakamura for help with DNA fingerprinting, H. Kishikawa, T. Kasai and R. Kleinfeld for assistance in preparing this manuscript, and J. Eppig for help and advice.

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Authors and Affiliations

  1. Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, 96822, Hawaii, USA

    T. Wakayama, A. C. F. Perry, M. Zuccotti & R. Yanagimachi

  2. Department of Veterinary Anatomy, Faculty of Agriculture, University of Tokyo, Bunkyo-ku, 113, Tokyo, Japan

    T. Wakayama

  3. Department of Signalling, Babraham Institute, Cambridge, CB2 4AT, UK

    A. C. F. Perry

  4. Dipartimento Biologia Animale, Laboratorio Biologia dello Sviluppo, University of Pavia, Piazza Botta 10, Pavia, 27100, Italy

    M. Zuccotti

  5. Jackson Laboratory, 600 Main Street, Bar Harbor, 04609, Maine, USA

    K. R. Johnson

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Wakayama, T., Perry, A., Zuccotti, M. et al. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–374 (1998). https://doi.org/10.1038/28615

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