Comparison of different fertilisation media for an in vitro maturation-fertilisation-culture system using flow-cytometrically sorted X chromosome-bearing spermatozoa for bovine embryo production

Abstract

High demand exists among commercial cattle producers for in vitro-derived bovine embryos fertilised with female sex-sorted spermatozoa from high-value breeding stock. The aim of this study was to evaluate three fertilisation media, namely M199, synthetic oviductal fluid (SOF) and Tyrode's albumin-lactate-pyruvate (TALP), on IVF performance using female sex-sorted spermatozoa. In all, 1143, 1220 and 1041 cumulus-oocyte complexes were fertilised in M199, SOF and TALP, respectively. There were significant differences among fertilisation media (P<0.05) in cleavage rate (M199≤57%, SOF≤71% and TALP≤72%), blastocyst formation (M199≤9%, SOF≤20% and TALP≤19%), proportion of Grade 1 blastocysts (M199≤15%, SOF≤52% and TALP≤51%), proportion of Grade 3 blastocysts (M199≤58%, SOF≤21% and TALP≤20%) and hatching rates (M199≤29%, SOF≤60% and TALP≤65%). The inner cell mass (ICM) and trophectoderm (TE) cells of Day 7 blastocysts were also affected by the fertilisation medium. Embryos derived from SOF and TALP fertilisation media had higher numbers of ICM, TE and total cells than those fertilised in M199. In conclusion, fertilisation media affected cleavage rate, as well as subsequent embryo development, quality and hatching ability. SOF and TALP fertilisation media produced significantly more embryos of higher quality than M199.