title: Embryo Aggregation in Pig Improves Cloning Efficiency and Embryo Quality.
abstract: In this study, we analyzed the effects of the cloned embryo aggregation on in vitro embryo development and embryo quality by measuring blastocyst diameter and cell number, DNA fragmentation levels and the expression of genes associated with pluripotency, apoptosis, trophoblast and DNA methylation in the porcine. Zona-free reconstructed cloned embryos were cultured in the well of the well system, placing one (1x non aggregated group) or three (3x group) embryos per microwell. Our results showed that aggregation of three embryos increased blastocyst formation rate and blastocyst diameter of cloned pig embryos. DNA fragmentation levels in 3x aggregated cloned blastocysts were significantly decreased compared to 1x blastocysts. Levels of Oct4, Klf4, Igf2, Bax and Dnmt 1 transcripts were significantly higher in aggregated embryos, whereas Nanog levels were not affected. Transcripts of Cdx2 and Bcl-xl were essentially non-detectable. Our study suggests that embryo aggregation in the porcine may be beneficial for cloned embryo development and embryo quality, through a reduction in apoptotic levels and an improvement in cell reprogramming.
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: Carla Paola
authors.initials: CP
authors.lastname: Buemo
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: Andrés
authors.initials: A
authors.lastname: Gambini
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: Lucia Natalia
authors.initials: LN
authors.lastname: Moro
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: María Inés
authors.initials: MI
authors.lastname: Hiriart
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: Rafael
authors.initials: R
authors.lastname: Fernández-Martín
authors.affiliation: Department of Molecular Medicine, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, and Norwegian Center for Stem Cell Research, Oslo, Norway.
authors.forename: Philippe
authors.initials: P
authors.lastname: Collas
authors.affiliation: Laboratory of Animal Biotechnology, Faculty of Agriculture, University of Buenos Aires, Buenos Aires, Argentina.|National Institute of Scientific and Technological Research, Buenos Aires, Argentina.
authors.forename: Daniel Felipe
authors.initials: DF
authors.lastname: Salamone
mesh_terms: D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D065150:Cellular Reprogramming / Q000235:genetics; D019976:Cloning, Organism; D053938:DNA Fragmentation; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian; D047108:Embryonic Development; D015870:Gene Expression; D020869:Gene Expression Profiling; D013552:Swine; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D014157:Transcription Factors