title: Growth factors and growth hormone enhance in vitro embryo production and post-thaw survival of vitrified bovine blastocysts.
abstract: The objective of this study was to assess the influence of specific growth factors and growth hormone (GH) in the culture medium on in vitro embryo production and post-thaw survival of vitrified blastocysts. In total, 1673 bovine oocytes were used for evaluating the nuclear status of the oocytes after in vitro maturation (n=560) or for in vitro fertilization (IVF, n=1113) and distributed in five treatment groups: (1). medium only control; (2). activin (10 ng/ml); (3). epidermal growth factor (EGF) (10 ng/ml); (4). insulin 5 microg/ml and (5). GH (100 ng/ml). There was an increase (P<0.05 and P<0.01, respectively) in the percentage of oocytes that reached meta phase II, developed to blastocysts and hatched, as well as in the blastocyst cell number in the groups treated with activin, EGF and GH compared to controls. There was no significant difference between insulin and control groups. A total of 465 blastocysts were vitrified in a three-step protocol using ethylene glycol and polyvinylpyrrolidone. After thawing, embryos were cultured in five treatments groups as described above. Groups EGF and GH had higher (P<0.05) survival rates with a mean blastocyst survival of 95.0+/-1.5 and 93.1+/-3.5%, respectively, while mean hatching rate was higher for EGF and activin groups (75.3+/-3.4 and 62.0+/-3.2%, respectively). Thawed control blastocysts had a mean cell count of 52.7+/-3.3%. With the exception of insulin, all growth factors and GH tested showed higher (P<0.01) total cell numbers when compared to controls. In conclusion, addition of growth factors and GH in the culture media has favorable effects on in vitro maturation, in vitro embryo production, and post-thaw survival of vitrified blastocysts.
authors.affiliation: Laboratory of Animal Reproduction and Applied Biotechnology, The United Graduate School of Veterinary Sciences, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: N R
authors.initials: NR
authors.lastname: Mtango
authors.forename: M D
authors.initials: MD
authors.lastname: Varisanga
authors.forename: Y J
authors.initials: YJ
authors.lastname: Dong
authors.forename: R
authors.initials: R
authors.lastname: Rajamahendran
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D028341:Activins / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D013006:Growth Hormone / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008297:Male; D009865:Oocytes / Q000502:physiology; D011897:Random Allocation
chemical_list: D007328:Insulin; D028341:Activins; D004815:Epidermal Growth Factor; D013006:Growth Hormone