Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning

Cristiano Feltrin, Luís Henrique de Aguiar, Carlos Enrique Méndez Calderón, Igor de Sá Carneiro, Felipe de Jesus Moraes Junior, Josiane da Silva Quetz, Aldo ângelo Moreira Lima, Matthew Wheeler, Davide Rondina, José Luiz Rodrigues, James Donald Murray, Elizabeth Anne Maga, Luciana Relly Bertolini, Marcelo Bertolini

Research output: Contribution to journalArticle

Abstract

The birth of cloned goats has been well documented, but the overall goat cloning efficiency by somatic cell nuclear transfer procedures is still low, which may be further intensified in extreme environments. The aim of this study was to produce cloned goats under the conditions of the Brazilian Semi-Arid region, in a transgenic program for the expression of human lysozyme in the milk to target childhood diarrhea and malnutrition, comparing the effects of oocyte source, cell type, and embryo reconstruction procedures on in vitro and in vivo embryo survival after cloning by micromanipulation or by handmade cloning. The use of in vitro-matured oocytes resulted in more viable embryos after cloning than in vivo-matured cytoplasts, but no differences in pregnancy rates on day 23 were seen between oocyte sources (77.5 vs. 77.8%, respectively). The presence or absence of the zona pellucida for embryo reconstruction (78.8 vs. 76.0%, respectively) did not affect pregnancy outcome after transfer. However, pregnancy rate on day 23 was higher for embryos chemically activated by a conventional than a modified protocol (88.1 vs. 50.0%), and for embryos reconstructed with mesenchymal stem cells and fetal fibroblasts (100.0 and 93.3%) than with adult fibroblasts (64.7%). Although most pregnancies were lost, the birth of a cloned female was obtained from embryos reconstructed by micromanipulation using non-transgenic control cells and in vitro-matured oocytes with intact zona pellucida, after conventional activation and transfer at the 1-cell stage.

Original languageEnglish (US)
Pages (from-to)1110-1123
Number of pages14
JournalAnimal Reproduction
Volume14
Issue number4
DOIs
StatePublished - 2017

Keywords

  • Goats
  • Oocyte maturation
  • Somatic cell nuclear transfer
  • Zona-free embryos

ASJC Scopus subject areas

  • Animal Science and Zoology
  • veterinary(all)

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    Feltrin, C., de Aguiar, L. H., Calderón, C. E. M., Carneiro, I. D. S., Moraes Junior, F. D. J., Quetz, J. D. S., Lima, A. Â. M., Wheeler, M., Rondina, D., Rodrigues, J. L., Murray, J. D., Maga, E. A., Bertolini, L. R., & Bertolini, M. (2017). Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning. Animal Reproduction, 14(4), 1110-1123. https://doi.org/10.21451/1984-3143-AR0002