title: Dynamic medium containing kit ligand and follicle-stimulating hormone promotes follicular survival, activation, and growth during long-term in vitro culture of caprine preantral follicles.
abstract: The aim of this study was to evaluate the effects of a dynamic medium containing kit ligand (KL) and follicle-stimulating hormone (FSH) on the in vitro culture of caprine preantral follicles for 16 days. Ovarian fragments were cultured in α-MEM(+) containing or not containing KL (50 ng/ml) and/or FSH (50 ng/ml) added during the first (days 0-8) and/or second half (days 8-16) of the culture period. Noncultured (control) and cultured fragments were processed for histological and ultrastructural evaluation. After 1 day of culture, only the treatments performed with KL or FSH maintained a percentage of normal follicles similar to that of the control. After 16 days, all treatments using KL until day 8 (KL/KL, KL/FSH, and KL/FSH+KL) and only FSH during the entire culture period (FSH/FSH) showed higher rates of follicular survival compared to α-MEM(+) alone. After 1 and 8 days, the treatments initially cultured with KL increased the percentage of follicular activation in comparison to α-MEM(+) alone and other treatments. The highest follicular diameter after 16 days was observed in follicles cultured with KL until day 8 followed by FSH (KL/FSH). Furthermore, this treatment promoted, as early as after 1 day of culture, an increase in oocyte growth compared to α-MEM(+) alone. Ultrastructural analysis confirmed the integrity of follicles cultured in KL/FSH after 16 days. In conclusion, a dynamic medium containing KL and FSH maintained follicular integrity and promoted follicular activation and growth during the long-term in vitro culture of caprine preantral follicles.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, Brazil. dora.uece@gmail.com
authors.forename: I M T
authors.initials: IM
authors.lastname: Lima
authors.forename: J J H
authors.initials: JJ
authors.lastname: Celestino
authors.forename: L R
authors.initials: LR
authors.lastname: Faustino
authors.forename: D M
authors.initials: DM
authors.lastname: Magalhães-Padilha
authors.forename: R
authors.initials: R
authors.lastname: Rossetto
authors.forename: I R
authors.initials: IR
authors.lastname: Brito
authors.forename: M A M
authors.initials: MA
authors.lastname: Donato
authors.forename: C A P
authors.initials: CA
authors.lastname: Lopes
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
authors.forename: C A
authors.initials: CA
authors.lastname: Peixoto
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.forename: A P R
authors.initials: AP
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D003470:Culture Media; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006041:Goats / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology*
chemical_list: D003470:Culture Media; D019089:Stem Cell Factor; D005640:Follicle Stimulating Hormone