236 effect of cyclic adenosine monophosphate modulator regulators in association with bmp15 on bovine embryo development in vitro

Proceedings of the Annual Conference of the International Embryo Transfer Society, Versailles, France, 10–13 January 2015BMP15 is a promising peptide to improve oocyte competence; also, addition of cyclic adenosine monophosphate modulator (cAMP) regulators prevents spontaneous maturation in vitro and promotes embryo development. We aimed to assess embryo development after prematuration [pre-in vitro maturation (IVM)] with IBMX and Forskolin (FSK) and maturation in the presence or absence of a purified pro mature region of BMP15. Immature cumulus-oocyte complexes (COC) were cultured in vitroMat (IVF Vet Solutions, Adelaide, Australia) plus 4mgmL(-1) fatty acid free-BSA and rhFSH (0.1IUmL(-1)), then divided into the following treatment groups: 1) spontaneous IVM: 24h of IVM; 2) spontaneous IVM+BMP15: 24h of IVM in the presence of BMP15 (100ngmL(-1)); 3) Pre 2 h: pretreatment with IBMX (500µM; Sigma-Aldrich) and FSK (100µM; Sigma-Aldrich) for 2h following 24h maturation; and 4) Pre 2 h+BMP15: pretreatment with IBMX and FSK for 2h following 24h maturation in the presence of BMP15 (100ngmL(-1)). After maturation, oocytes were inseminated and zygotes were cultured for 5 days in VitroCleave (IVF Vet Solutions, Adelaide, Australia) and transferred into VitroBlast (IVF Vet Solutions, Adelaide, Australia) until blastocyst assessment (Days 7 and 8). Zona-intact embryos were retrieved to assess differential staining of trophectoderm and inner cell mass. Data were transformed into a logarithm and analysed by 1-way ANOVA and post hoc least significant difference using SigmaStat software (SPSS Inc., San Jose, CA, USA; P<0.05). There was no difference among groups on cleavage rates or blastocyst rates at Day 7; however, both Pre 2h treatments increase hatched blastocyst rates at Day 8 of embryo development (Table 1). Supplementation with BMP15 increased total blastocyst rates at Day 8, regardless of pretreatment with IBMX+FSK (Table 1). Our data demonstrate that embryos from oocytes matured in the presence of BMP15 or pretreated with IBMX+FSK increase trophectoderm and total cell numbers; however, no differences were observed for inner cell mass. We conclude that Pre 2h treatment or BMP15 increase embryo development; however, no effect of cAMP regulators in association with BMP15 on embryo development was observed.M. F. Machado, M. F. G. Nogueira, R. B. Gilchrist, M. L. Sutton-McDowall, D. G. Mottershead, M. A. White and J. G. Thompso