title: Rutin can replace the use of three other antioxidants in the culture medium, maintaining the viability of sheep isolated secondary follicles.
abstract: The present study evaluated the effect of addition of rutin alone or combined with other antioxidants (transferrin, selenium and ascorbic acid) present in the culture medium on the in vitro development of ovine isolated secondary follicles. After collection of the sheep ovaries, secondary follicles (200-230 μm) were isolated and cultured for 12 days in α-Minimal Essential Medium (α-MEM) supplemented with BSA, insulin, glutamine and hypoxanthine (α-MEM: antioxidant free-medium) or in this medium also added by transferrin, selenium and ascorbic acid (AO: base medium with antioxidants). Moreover, different concentrations of rutin (0.1; 1 or 10 μg/mL) were added to the different base media (α-MEM or AO). The parameters analyzed were morphology, antrum formation, extrusion rate, follicular diameter, growth and fully-grown oocytes (oocytes ≥ 110 μm) rates. In treatments that had the best results of morphology, follicular viability, apoptosis, glutathione (GSH), reactive oxygen species (ROS) levels and mitochondrial activity were also analyzed. After 12 days, the percentage of normal follicles was higher (P < 0.05) in α-MEM + 0.1 μg/mL rutin than the other treatments, except compared to AO medium (P > 0.05). There is no difference (P > 0.05) in the diameter and growth rate among treatments. Moreover, AO medium and α-MEM + 0.1 μg/mL rutin showed similar (P > 0.05) percentages of follicular viability, antrum formation, extruded follicles, fully-grown oocytes, levels of ROS and active mitochondria. However, α-MEM + 0.1 μg/mL rutin treatment showed higher (P > 0.05) GSH levels than AO medium. In conclusion, 0.1 μg/mL rutin can be used as the single antioxidant present in the base medium, replacing the addition of transferrin, selenium and ascorbic acid during in vitro culture of ovine secondary follicles, maintaining follicular viability and increasing GSH levels.
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: T L B G
authors.initials: TLBG
authors.lastname: Lins
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: A Y P
authors.initials: AYP
authors.lastname: Cavalcante
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: J M S
authors.initials: JMS
authors.lastname: Santos
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: V G
authors.initials: VG
authors.lastname: Menezes
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: V R P
authors.initials: VRP
authors.lastname: Barros
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: R S
authors.initials: RS
authors.lastname: Barberino
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: M É S
authors.initials: MÉS
authors.lastname: Bezerra
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil.
authors.forename: T J S
authors.initials: TJS
authors.lastname: Macedo
authors.affiliation: Nucleus of Biotechnology Applied to Ovarian Follicle Development, Federal University of San Francisco Valley, 56300-990, Petrolina, PE, Brazil. Electronic address: helena.matos@univasf.edu.br.
authors.forename: M H T
authors.initials: MHT
authors.lastname: Matos
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis; D001205:Ascorbic Acid / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000662:veterinary; D002470:Cell Survival; D003470:Culture Media; D005260:Female; D005979:Glutathione Peroxidase / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D008928:Mitochondria / Q000378:metabolism / Q000502:physiology; D006080:Ovarian Follicle / Q000187:drug effects* / Q000254:growth & development; D017382:Reactive Oxygen Species / Q000378:metabolism; D012431:Rutin / Q000494:pharmacology*; D012643:Selenium / Q000494:pharmacology; D012756:Sheep; D014168:Transferrin / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D003470:Culture Media; D017382:Reactive Oxygen Species; D014168:Transferrin; D012431:Rutin; D005979:Glutathione Peroxidase; D012643:Selenium; D001205:Ascorbic Acid
keywords: Flavonoid; GSH; Oocyte; Ovine; Survival