Abstract
The effect of resveratrol supplementation on fresh (E1) or vitrified/warmed (E2) in vitro produced bovine embryos was investigated by evaluating the time-dependent response. After in vitro production, resveratrol (0.5 µM) was added to the incubation media and after two incubation periods with or without resveratrol, blastocysts were re-cultured for 24h. The rates of re-expansion, hatching, total cell number (TCN), apoptotic cells (ACN), reactive oxygen species (ROS) and intracellular glutathione (GSH) content were evaluated. For E1, the re-expansion rate differed at 6 and 10h within and between treatments (P<0.05), as did the re-expansion rate after 24h (P<0.01). The hatching rate increased after 10h with resveratrol (P<0.01) with differences within (P<0.05), but not between treatments after 24h of re-cultivation. At E2, hatching rate differed between treatments at 24h (P<0.01), with higher TCN in resveratrol-treated blastocysts after 10h (P<0.01). Resveratrol supplementation reduced ROS generation in E1 and E2 after 10h of incubation and increased GSH content (P<0.01). These results indicate that supplementation of holding re-cultivation medium with resveratrol for treatment of fresh or vitrified/warmed in vitro produced bovine embryos has a positive and time-dependent effect. The reduction of ROS content, the increase of GSH and the anti-apoptotic ability of resveratrol are responsible for its protective effects, allowing an extension of embryo storage time before transfer to recipients.
Highlights
The increase in human population and the demand for food in the world has served as a stimulus for producers to seek alternatives to increase production over time, and in this context, the bovine in vitro embryo production (IVP) industry has grown considerably worldwide (Sanches et al, 2019)
During the entire IVP process, the amount of reactive oxygen species (ROS) might imbalance the availability of antioxidants as ROS are produced as a result of cellular metabolism; ROS levels play a critical role in the success of in vitro fertilization because the quality of the embryo could be impaired owing to harmful effects of the culture environment
Embryos incubated for 6 h before the re-cultivation of up to 24 h presented a higher hatched rate (Control, 43.3% and resveratrol 0.5 μM (Resv), 48.4%) at the final time than those incubated for 10 h (Control, 25.0% and Resv, 27.2%; P < 0.05)
Summary
The increase in human population and the demand for food in the world has served as a stimulus for producers to seek alternatives to increase production over time, and in this context, the bovine in vitro embryo production (IVP) industry has grown considerably worldwide (Sanches et al, 2019). During the entire IVP process, the amount of ROS might imbalance the availability of antioxidants as ROS are produced as a result of cellular metabolism; ROS levels play a critical role in the success of in vitro fertilization because the quality of the embryo could be impaired owing to harmful effects of the culture environment. The increase in cellular damage because of increased ROS production during the IVP process is well documented (Lee et al, 2012), as well the benefits of adding antioxidant molecules to supplement culture media and decrease ROS production to improve the quality and to protect embryos against damage to DNA and other biomolecules, raising their developmental competence (Morado et al, 2009)
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