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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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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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