The objective of this study was to investigate the effect of VEGF and Cysteamine during in vitro maturation (IVM) of bovine oocytes on GSH content and developmental competence. For this purpose, experiments were designed to evaluate the effect of 0, 100, 300, and 500 ng/mL VEGF in IVM medium on: GSH content in oocytes and cumulus cells (Exp. 1) and subsequent embryo development (Exp. 2). Also, influence of adding 500 ng/mL VEGF and 100 μM Cysteamine to IVM medium on GSH content in oocytes and cumulus cells (Exp. 3) and oocyte developmental capacity (Exp. 4) were evaluated. Oocytes were matured in: a) Control; b) VEGF 0-3 h; c) Cysteamine 4-24 h; d) VEGF 0-3 h + Cysteamine 4-24 h; and e) VEGF + Cysteamine 24 h. The results showed that: i) VEGF did not alter GSH content in oocytes and cumulus cells; (ii) supplementation of 300 and 500 ng/mL VEGF increased blastocyst yield; (iii) the presence of VEGF + Cysteamine simultaneously during 24 h improved GSH content but not embryo development; and (iv) the presence of VEGF during the first 3 h + Cysteamine from 4 to 24 h increased GSH concentrations and subsequent embryo development. In conclusion, the addition of VEGF and Cysteamine in two sequential steps to maturation medium result in an improvement of cytoplasmic maturation, with a positive impact on oocyte developmental capacity by increasing the efficiency of in vitro blastocyst production. However, the effect was detrimental when both VEGF and Cysteamine were present during 24 of IVM.

The aim of this study was to investigate the influence of copper (Cu) during in vitro maturation (IVM) on apoptosis and DNA integrity of cumulus cells (CC); and oocyte viability. Also, the role of CC in the transport of Cu during IVM was evaluated on oocyte developmental capacity. Damage of DNA was higher in CC matured without Cu (0 µg/dl Cu, P < 0.01) with respect to cells treated with Cu for cumulus-oocyte complexes (COCs) exposed to 0, 20, 40, or 60 µg/dl Cu). The percentage of apoptotic cells was higher in CC matured without Cu than in CC matured with Cu. Cumulus expansion and viability of CC did not show differences in COC treated with 0, 20, 40, or 60 µg/dl Cu during IVM. After in vitro fertilization (IVF), cleavage rates were higher in COC and DO + CC (denuded oocytes + CC) with or without Cu than in DO. Independently of CC presence (COC, DO + CC or DO) the blastocyst rates were higher when 60 µg/dl Cu was added to IVM medium compared to medium alone. These results indicate that Cu supplementation to IVM medium: (i) decreased DNA damage and apoptosis in CC; (ii) did not modify oocyte viability and cumulus expansion; and (iii) improved subsequent embryo development up to blastocyst stage regardless of CC presence during IVM.