Developmental competence and epigenetic profile of porcine embryos produced by two different cloning methods

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DOI

  • Ying Liu
  • ,
  • Andrea Lucas-Hahn, Institute of Farm Animal Genetics (FLI), Germany
  • Bjoern Petersen, Institute of Farm Animal Genetics, FLI, Germany
  • Rong Li
  • ,
  • Doris Hermann, Institute for Farm Animal Genetics, Germany
  • Petra Hassel, Institute for Farm Animal Genetics, Germany
  • Maren Ziegler, Institute for Farm Animal Genetics, Germany
  • Knud Erik Larsen
  • Heiner Niemann, Institute of Farm Animal Genetics, FLI, Germany
  • Henrik Callesen
The “Dolly” based cloning (classical nuclear transfer, [CNT]) and the handmade cloning (HMC) are methods that are nowadays routinely used for somatic cloning of large domestic species. Both cloning protocols share several similarities, but differ with regard to the required in vitro culture, which in turn results in different time intervals until embryo transfer. It is not yet known whether the differences between cloned embryos from the two protocols are due to the cloning methods themselves or the in vitro culture, as some studies have shown detrimental effects of in vitro culture on conventionally produced embryos. The goal of this study was to unravel putative differences between two cloning methods, with regard to developmental competence, expression profile of a panel of developmentally important genes and epigenetic profile of porcine cloned embryos produced by either CNT or HMC, either with (D5 or D6) or without (D0) in vitro culture. Embryos cloned by these two methods had a similar morphological appearance on D0, but displayed different cleavage rates and different quality of blastocysts, with HMC embryos showing higher blastocyst rates (HMC vs. CNT: 35% vs. 10%, p < 0.05) and cell numbers per blastocyst (HMC vs. CNT: 31 vs. 23 on D5 and 42 vs. 18 on D6, p < 0.05) compared to CNT embryos. With regard to histone acetylation and gene expression, CNT and HMC derived cloned embryos were similar on D0, but differed on D6. In conclusion, both cloning methods and the in vitro culture may affect porcine embryo development and epigenetic profile. The two cloning methods essentially produce embryos of similar quality on D0 and after 5 days in vitro culture, but thereafter both histone acetylation and gene expression differ between the two types of cloned embryos.
Original languageEnglish
JournalCellular Reprogramming
Volume19
Issue3
Pages (from-to)171-179
Number of pages9
ISSN2152-4971
DOIs
Publication statusPublished - Jun 2017

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