Embryo Aggregation in Pig Improves Cloning Efficiency and Embryo Quality

GAMBINI A; BUEMO CP; MORO L.N; COLLAS P; HIRIART M.I; FERNANDEZ Y MARTIN R; SALAMONE D.F

PLOS ONE

PUBLIC LIBRARY SCIENCE

Lugar: San Francisco; Año: 2016 vol. 11

1932-6203

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 withpluripotency, apoptosis, trophoblast and DNA methylation in the porcine. Zona­free reconstructed cloned embryos were cultured inthe well of the well system, placing one (1x non aggregated group) or three (3x group) embryos per microwell. Our results showedthat aggregation of three embryos increased blastocyst formation rate and blastocyst diameter of cloned pig embryos. DNAfragmentation 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 Bcl­xl were essentially non­detectable. Our study suggests that embryo aggregation in the porcine may bebeneficial for cloned embryo development and embryo quality, through a reduction in apoptotic levels and an improvement in cellreprogramming.