title: Effect of resveratrol supplementation during culture on the quality and cryotolerance of bovine in vitro produced embryos.
abstract: The aim of the study was to evaluate whether resveratrol supplementation of bovine culture medium improves in vitro blastocyst development, embryo cryotolerance and cell numbers. Abattoir-derived oocytes were matured and fertilized in vitro according to standard procedure. Twenty hours after IVF, zygotes were cultured in SOF medium, supplemented with 0 (control, n=439), 0.25μM (n=422), 0.5μM (n=447) and 1μM resveratrol (n=416). On Day 7 (IVF=Day 0) blastocysts were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide and 0.5M sucrose. Development rate, i.e. the percentage of embryos resuming development to reach a more advanced stage, and hatching rate were evaluated after 24 and 48h culture. Blastocysts cultured with (0.5μM) and without resveratrol underwent differential staining to count inner cell mass (ICM) and trophectoderm (TE) cells. Resveratrol during culture did not increase blastocyst yields (57.1, 57.7, 59.2 and 46.6%, respectively in 0, 0.25, 0.5 and 1μM resveratrol). However, 0.5μM resveratrol improved embryo cryotolerance compared to the control, as indicated by higher development rates (67.3% vs 50.3%, respectively; P<0.01) and hatching rates (58.9% vs 30.9%, respectively; P<0.01) recorded after 48h post-warming culture. Blastocysts produced in the control and in 0.5μM resveratrol groups had similar numbers of ICM (34.1 and 36.4, respectively), TE (88.1 and 85.3, respectively) and total (122.2 and 121.7, respectively) cells. In conclusion, low levels of resveratrol during in vitro culture improve the quality of IVP bovine embryos, as indicated by their increased resistance to cryopreservation.
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Salzano
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Albero
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Zullo
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy. Electronic address: neglia@unina.it.
authors.forename: G
authors.initials: G
authors.lastname: Neglia
authors.affiliation: Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt.
authors.forename: A
authors.initials: A
authors.lastname: Abdel-Wahab
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Bifulco
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Zicarelli
authors.affiliation: Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Gasparrini
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002417:Cattle / Q000196:embryology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000662:veterinary; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods* / Q000592:standards / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects; D005307:Fertilization in Vitro / Q000662:veterinary; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*; D013997:Time Factors; D058989:Vitrification / Q000187:drug effects
chemical_list: D013267:Stilbenes; D000077185:Resveratrol
keywords: Bovine; Cryotolerance; Embryo culture; Resveratrol