title: In vitro development of bovine secondary follicles in two- and three-dimensional culture systems using vascular endothelial growth factor, insulin-like growth factor-1, and growth hormone.
abstract: The aim of this study was to evaluate the development and estradiol production of isolated bovine secondary follicles in two-dimensional (2D, experiment 1) and three-dimensional (3D using alginate, experiment 2) long-term culture systems in the absence (control group; only α-MEM(+)) or presence of vascular endothelial growth factor (VEGF), insulin-like growth factor-1, or GH alone, or a combination of all. A total of 363 isolated secondary follicles were cultured individually for 32 days at 38.5 °C in 5% CO2 in a humidified incubator with addition of medium (5 μL) every other day. In 2D culture system, follicular growth and antrum formation rates were higher (P < 0.05) in VEGF treatment compared with the other treatments. In 3D culture system, only estradiol concentration was greater (P < 0.05) in the GH than in the control group, whereas the other end points were similar (P > 0.05). In summary, this study demonstrated that the benefits of using a certain type of medium supplement depended on the culture system (2D vs. 3D). Vascular endothelial growth factor was an effective supplement for the in vitro culture of bovine secondary follicles when the 2D culture system was used, whereas GH only affected estradiol production using the 3D culture system. This study sheds light on advancements in methodology to facilitate subsequent studies on bovine preantral follicle development.
authors.affiliation: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA; Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, Ceará, Brazil.
authors.forename: V R
authors.initials: VR
authors.lastname: Araújo
authors.affiliation: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA.
authors.forename: M O
authors.initials: MO
authors.lastname: Gastal
authors.affiliation: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA.
authors.forename: A
authors.initials: A
authors.lastname: Wischral
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, Ceará, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.affiliation: Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, Illinois, USA. Electronic address: egastal@siu.edu.
authors.forename: E L
authors.initials: EL
authors.lastname: Gastal
mesh_terms: D000818:Animals; D002417:Cattle / Q000502:physiology*; D018929:Cell Culture Techniques / Q000662:veterinary; D005260:Female; D013006:Growth Hormone / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000254:growth & development*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
chemical_list: D042461:Vascular Endothelial Growth Factor A; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone
keywords: Cattle; Culture systems; Estradiol; Growth factors; Preantral follicles