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Cloned pigs produced by nuclear transfer from adult somatic cells

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.

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Figure 1: Representation of the double nuclear transfer procedure for the production of viable piglets using cultured adult somatic granulosa cells as nuclear donors.
Figure 2: A litter of five live piglets derived by nuclear transfer using cultured adult granulosa cells as nuclear donors.

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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).

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Correspondence to Irina A. Polejaeva.

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Polejaeva, I., Chen, SH., Vaught, T. et al. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407, 86–90 (2000). https://doi.org/10.1038/35024082

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