title: In vitro development of secondary follicles from pre-pubertal and adult goats cultured in two-dimensional or three-dimensional systems.
abstract: The aim of this study was to evaluate the influence of two-dimensional (2D) and three-dimensional (3D) alginate culture systems on in vitro development of pre-antral caprine follicles. In addition, the influence of the reproductive age of the ovary donor on the in vitro culture success was investigated. Pre-antral follicles from pre-pubertal or adult goats were isolated and cultured directly on a plastic surface (2D) or encapsulated in an alginate-based matrix (3D). After 18 days, the oocytes underwent in vitro maturation (IVM) and in vitro fertilization (IVF) to produce embryos. The 3D system showed higher rates of follicle survival, lower rates of oocyte extrusion, and a greater number of recovered oocytes for IVM and IVF (P < 0.05). Only pre-antral follicles from adult animals produced MII oocytes and embryos. The estradiol concentrations increased from day 2 to day 12 of culture in all groups tested (P < 0.05). Conversely, progesterone concentrations were lower in 3D-cultured follicles than in 2D-cultured follicles, with differences on days 2 and 6 of culture (P < 0.05). We provide compelling evidence that a 2D or 3D alginate in vitro culture system offers a promising approach to achieving full in vitro development of caprine pre-antral follicles to produce mature oocytes that are capable of fertilization and viable embryos.
authors.affiliation: Programa de Pós-Graduação em Ciências Veterinárias (PPGCV),Laboratório de Manipulação de Oócitos e Folículos Pré-Antrais (LAMOFOPA),Universidade Estadual do Ceará (UECE),Av. Paranjana 1700,Campus do Itaperi,CEP: 60740-903,Fortaleza - CE -Brasil.
authors.forename: G M
authors.initials: GM
authors.lastname: Silva
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
authors.forename: R
authors.initials: R
authors.lastname: Rossetto
authors.affiliation: Health Sciences Center,University of Fortaleza,Fortaleza - CE,Brazil.
authors.forename: R N
authors.initials: RN
authors.lastname: Chaves
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
authors.forename: A B G
authors.initials: AB
authors.lastname: Duarte
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
authors.forename: V R
authors.initials: VR
authors.lastname: Araújo
authors.affiliation: Molecular Biology and Development Laboratory,University of Fortaleza,Fortaleza,Ceará,Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Feltrin
authors.affiliation: Human Reproduction Division,Department of Obstetrics and Gynecology,Faculty of Medicine of Ribeirao Preto,University of São Paulo,Ribeirão Preto - SP,Brazil.
authors.forename: M P
authors.initials: MP
authors.lastname: Bernuci
authors.affiliation: Department of Morphology,Stomatology and Physiology,Faculty of Dentistry of Ribeirão Preto,University of São Paulo,Ribeirão Preto - SP,Brazil.
authors.forename: J A
authors.initials: JA
authors.lastname: Anselmo-Franci
authors.affiliation: Northwestern University,Department of Obstetrics and Gynecology,Chicago,IL,USA.
authors.forename: M
authors.initials: M
authors.lastname: Xu
authors.affiliation: Northwestern University,Department of Obstetrics and Gynecology,Chicago,IL,USA.
authors.forename: T K
authors.initials: TK
authors.lastname: Woodruff
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles,Faculty of Veterinary Medicine,State University of Ceará,Fortaleza - CE,Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
mesh_terms: D000367:Age Factors; D000464:Alginates; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002470:Cell Survival; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D020723:Glucuronic Acid; D006041:Goats; D006603:Hexuronic Acids; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D006080:Ovarian Follicle / Q000254:growth & development* / Q000502:physiology; D011627:Puberty
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D004958:Estradiol; D020723:Glucuronic Acid
keywords: Alginate; Follicle culture; Oocyte; Pre-antral follicle; Steroid secretion