title: Cysteamine supplementation during in vitro maturation of slaughterhouse- and opu-derived bovine oocytes improves embryonic development without affecting cryotolerance, pregnancy rate, and calf characteristics.
abstract: Optimization of ovum pick up (OPU) followed by in vitro embryo production (IVP) is strongly driven by the needs of both beef and dairy cattle breeders to enhance genetic improvement. The rapidly growing use of genomic selection in cattle has increased the interest in using OPU-IVP technology to increase the number of embryos and offspring per donor, thus allowing enhanced selection intensity for the next generation. The aim of this study was to optimize embryo production through supplementation of cysteamine during in vitro maturation (IVM) and in vitro culture (IVC) of both slaughterhouse- and OPU-derived oocytes. The effects on embryo production and on embryo cryotolerance, post-transfer embryo survival, and calf characteristics, including gestation length, birth weight, perinatal mortality, and sex ratio were studied. In study 1, immature slaughterhouse-derived cumulus-oocyte complexes (COCs) were matured in IVM medium supplemented with or without 0.1 mM cysteamine, fertilized and cultured for 7 days in 0.5 ml SOFaaBSA. In study 2, cysteamine was present during both IVM (0.1 mM) and IVC (0.01, 0.05, 0.1 mM) from Days 1 to 4. In study 3, OPU-derived COCs were matured in medium supplemented with or without 0.1 mM cysteamine in a 2 × 2 factorial design (OPU week and cysteamine treatment). Embryos were evaluated for stage and grade on Day 7 and, depending on the number of transferable embryos and recipients available, the embryos were transferred either fresh or frozen-thawed at a later date. The presence of cysteamine during IVM significantly increased the embryo production rate with slaughterhouse-derived COCs (24.0% vs. 19.4%). The higher number of embryos at Day 7 was due to an increased number of blastocysts, whereas the distribution of embryos among different quality grades and cryotolerance was not affected. Embryo production rate was negatively affected when cysteamine was present during both the processes of IVM and IVC during Days 1 to 4 of culture (13.2%-19.3% vs. 26.4%). The presence of cysteamine during IVM of OPU-derived COCs also significantly increased the embryo production rate (34.4% vs. 23.4%). The higher number of embryos was again totally due to an increased number of blastocysts, whereas cryotolerance was not affected. The relative increase in embryo production rate was higher with OPU-derived oocytes compared with slaughterhouse-derived COCs (47% vs. 24%). This improvement resulted in a mean of 1.73 transferable embryos per OPU session compared with 1.06 in the absence of cysteamine. The presence of cysteamine did not affect pregnancy rate, gestation length, birth weight, perinatal mortality, and sex of calves born from either fresh or frozen-thawed embryos. This study reported that cysteamine supplementation during IVM greatly improved the efficiency and affectivity of an OPU-IVP program.
authors.affiliation: CRV BV, 6800 AL Arnhem, The Netherlands. sybrand.merton@crv4all.com
authors.forename: J S
authors.initials: JS
authors.lastname: Merton
authors.forename: H M
authors.initials: HM
authors.lastname: Knijn
authors.forename: H
authors.initials: H
authors.lastname: Flapper
authors.forename: F
authors.initials: F
authors.lastname: Dotinga
authors.forename: B A J
authors.initials: BA
authors.lastname: Roelen
authors.forename: P L A M
authors.initials: PL
authors.lastname: Vos
authors.forename: E
authors.initials: E
authors.lastname: Mullaart
mesh_terms: D000003:Abattoirs; D000818:Animals; D000831:Animals, Newborn / Q000254:growth & development / Q000502:physiology*; D002417:Cattle / Q000196:embryology / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D018592:Cross-Over Studies; D015925:Cryopreservation / Q000662:veterinary; D003543:Cysteamine / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology; D005260:Female; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary; D054315:Oocyte Retrieval / Q000379:methods / Q000662:veterinary; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D011247:Pregnancy; D018873:Pregnancy Rate
chemical_list: D003543:Cysteamine
keywords: Bovine; Culture; Cysteamine; Glutathione; OPU-IVP; Oxidative stress; ROS