Letter | Published:

Cloned pigs produced by nuclear transfer from adult somatic cells

Nature volume 407, pages 8690 (07 September 2000) | Download Citation

Subjects

Abstract

Since the first report of live mammals produced by nuclear transfer from a cultured differentiated cell population in 1995 (ref. 1), successful development has been obtained in sheep2,3, cattle4, mice5 and goats6 using a variety of somatic cell types as nuclear donors. The methodology used for embryo reconstruction in each of these species is essentially similar: diploid donor nuclei have been transplanted into enucleated MII oocytes that are activated on, or after transfer. In sheep2 and goat6 pre-activated oocytes have also proved successful as cytoplast recipients. The reconstructed embryos are then cultured and selected embryos transferred to surrogate recipients for development to term. In pigs, nuclear transfer has been significantly less successful; a single piglet was reported after transfer of a blastomere nucleus from a four-cell embryo to an enucleated oocyte7; however, no live offspring were obtained in studies using somatic cells such as diploid or mitotic fetal fibroblasts as nuclear donors8,9. The development of embryos reconstructed by nuclear transfer is dependent upon a range of factors. Here we investigate some of these factors and report the successful production of cloned piglets from a cultured adult somatic cell population using a new nuclear transfer procedure.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–67 ( 1996).

  2. 2.

    , , , & Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810–813 (1997).

  3. 3.

    , , & Production of cloned lambs from an established embryonic cell line: a comparison between in vivo- and in vitro-matured cytoplasts. Biol. Reprod. 57, 385–393 ( 1997).

  4. 4.

    et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 280, 1256– 1258 (1998).

  5. 5.

    , , , & Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–374 ( 1998).

  6. 6.

    et al. Production of goats by somatic cell nuclear transfer. Nature Biotechnol. 17, 456–461 (1999).

  7. 7.

    , & Nuclear transplantation in early pig embryos. Biol. Reprod. 41, 414–418 (1989).

  8. 8.

    et al. Development of pig embryos reconstructed by microinjection of cultured fetal fibroblast cells into in vitro matured oocytes. Anim. Reprod. Sci. 56, 133–141 ( 1999).

  9. 9.

    et al. Nuclear transplantation in pigs: M-phase karyoplast to M-phase cytoplast fusion. Reprod. Nutr. Dev. 36, 661–666 (1996).

  10. 10.

    Somatic cell nuclear transfer in mammals: Progress and applications. Cloning 1, 185–200 ( 2000)

  11. 11.

    , & The effect of reducing the number of embryos during early stages of gestation on the maintenance of pregnancy in the pig. J. Reprod. Fert. 12, 395–397 (1966).

  12. 12.

    et al. Gene transfer efficiency during gestation and the influence of co-transfer of non-manipulated embryos on production of transgenic mice. Transgenic Res. 3, 20– 25 (1994).

  13. 13.

    & Maintenance of unilateral pregnancy in the pig with induced corpora lutea. J. Anim. Sci. 32, 282–286 ( 1971).

  14. 14.

    et al. Artificial activation of porcine oocytes matured in vitro. Mol. Reprod. Dev. 28, 405– 409 (1991).

  15. 15.

    & Nuclear transplantation in the mouse embryo by microsurgery and cell fusion. Science 220 , 1300–1302 (1993).

  16. 16.

    & Nuclear transplantation in mouse embryos. J. Exp. Zool 228, 355– 362 (1993).

  17. 17.

    & Production of identical sextuplet mice by transferring metaphase nuclei from four-cell embryos. Proc. Natl Acad. Sci. USA 93, 13010–13013 (1996).

  18. 18.

    , & Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biol. Reprod. 60, 996–1005 (1999).

  19. 19.

    , , & Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev. Reprod. 1, 40–46 (1996).

  20. 20.

    et al. Homologous recombination in ovine somatic cells enables the production of gene targeted sheep by nuclear transfer. Nature 405, 1066–1069 (2000).

  21. 21.

    Xenotransplantation:recent progress & current perspectives. Curr. Opin. Immunol. 8, 721–728 (1996).

  22. 22.

    & Culture of pig embryos. J. Reprod. Fertil. Suppl. 48, 61 –73 (1993).

Download references

Acknowledgements

We thank B. Gragg, T. Akers, H. Bishop and J. McPherson for technical contributions to embryo production, embryo transfer and animal husbandry; P. Jobst for technical contributions in embryo production and nuclear transfer; W. Eyestone and K. Polson for valuable contributions in discussions; S. Pleasant and M. Crisman and their staff at the Virginia Maryland Regional Veterinary College for their critical roles in the Caesarian delivery and post-natal care of the cloned piglets. This research was funded in part by a grant from the National Institute of Science and Technology (NIST) Advanced Technology Program (ATP).

Author information

Author notes

    • Keith H. S. Campbell

    Present address: University of Nottingham, School of Biosciences, Sutton Bonington, Leicestershire LE12 5RN, UK

Affiliations

  1. *PPL Therapeutics Incorporated, 1700 Kraft Drive, Blacksburg, Virginia 24060, USA

    • Irina A. Polejaeva
    • , Shu-Hung Chen
    • , Todd D. Vaught
    • , Raymond L. Page
    • , June Mullins
    • , Suyapa Ball
    • , Yifan Dai
    • , Jeremy Boone
    • , Shawn Walker
    •  & David L. Ayares
  2. †PPL Therapeutics, Roslin , Midlothian EH25 9PP, UK

    • Alan Colman
    •  & Keith H. S. Campbell

Authors

  1. Search for Irina A. Polejaeva in:

  2. Search for Shu-Hung Chen in:

  3. Search for Todd D. Vaught in:

  4. Search for Raymond L. Page in:

  5. Search for June Mullins in:

  6. Search for Suyapa Ball in:

  7. Search for Yifan Dai in:

  8. Search for Jeremy Boone in:

  9. Search for Shawn Walker in:

  10. Search for David L. Ayares in:

  11. Search for Alan Colman in:

  12. Search for Keith H. S. Campbell in:

Corresponding author

Correspondence to Irina A. Polejaeva.

Supplementary information

Word documents

  1. 1.

    407086a0_S1.doc

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/35024082

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.