Separation of X-Bearing Ovine Sperm by Centrifugation in Discontinuous Ficoll Density Gradient: Effect on Gender of In Vitro Produced Embryos

Abstract

The aim of this study was the separation of X-bearing ovine sperm by centrifugation in discontinuous “Ficoll” density gradient and their effect on gender of in vitro produced embryos. The percentage of male and female ovine embryos produced after IVF which determined by polymerase chain reaction (PCR). The discontinuous Ficoll density gradient (three layer) centrifugation at 200×g (F1a) and 300×g (F 1b) showed a significant differences (P<0.05) in the sex ratio toward female embryos (63.6 and 54.5%) respectively. While the discontinuous Ficoll density gradient (four layer 80, 60, 40 and 20) centrifugation at 200×g (F2a) and 300×g (F2b) showed a significant differences (P<0.05) in the sex ratio toward female embryos (72.7 and 63.6%) respectively. While at four layer (80, 72, 64 and 50) concentration of ficoll the ratio was at 200×g (F3a) and 300×g (F3b) showed a significant differences (P<0.05) in the sex ratio toward female embryos (81.8, 72.7)%. From our results we can conclude that the purity of selected X-sperms and the percentage of female embryos obtained by IVF were good when we use F2a,2b and F3a,3b protocols than F1a,1b protocols, and the best result of IVF obtained when we apply high concentration of Ficoll media F3a,3b than F2a, 2b.