Handmade cloned and parthenogenetic goat embryos - A comparison of different culture media and donor cells

The aim of the present investigation was the production of handmade cloned and parthenogenetic goat embryos. One somatic cell was attached to an enucleated demi-oocyte and fused with another enucleated demi-oocyte with the aid of an electric pulse. Triplets were chemically activated with a Ca ionophore and 6-DMAP, and then cultured in three media. Blastocyst formation with fetal fibro cells was induced in RVCL (15.5 +/- 4.2%), mSOF (11.0 +/- 2.2%) and EDM (10.7 +/- 2.4%) media, respectively (P > 0.05). Similarly, adult fibroblast cells formation was also recorded in RVCL (14.1 +/- 3.3%), mSOF (9.6 +/- 2.7%) and EDM (9.7 +/- 2.2%) media, respectively (P > 0.05). PCR analysis of the highly polymorphic MHC class II DRB gene of cloned embryos and donor cells, recorded similar bands. Zona and zona-free parthenogenetic embryos were also produced with the aid of a Ca ionophore and 6-DMAP, following culture in the three media. Zona parthenogenetic hatched blastocysts were recorded in RVCL (6.8 +/- 0.9%), mSOF (1.2 +/- 0.7%) and EDM (5.5 +/- 0.7%) (P < 0.05) media, respectively. In the zona-free parthenogenetic blastocyst, formation was recorded in the RVCL (8.8 +/- 0.9%), mSOF (5.6 +/- 0.5%) and EDM (5.1 +/- 0.8%) (P < 0.05) media, respectively. In conclusion, cloned and parthenogenetic goat embryonic development was higher in the RVCL medium and cloned embryos production was similar with both types of donor cells in this medium. (C) 2012 Elsevier B.V. All rights reserved.