Culture of preantral ovarian follicles of Bos taurus indicus with alpha-lipoic acid

• Published in: Zygote (Cambridge) Vol. 30; no. 2; p. 206
• Main Authors: Bergamo, Larissa Zamparone, Bonato, Denis Vinicius, Bizarro-Silva, Camila, Bonato, Francieli Gesleine Capote, González, Suellen Miguez, Rossaneis, Ana Carolina, Verri, Waldiceu A, Morotti, Fábio, Seneda, Marcelo Marcondes
• Format: Journal Article
• Language: English
• Published: England 01-04-2022
• Subjects: Animals; Antioxidants - pharmacology; Cattle; Culture Media; Female; Ovarian Follicle; Ovary; Thioctic Acid - pharmacology; Tissue Culture Techniques; Antioxidants; In vitro culture of preantral follicles; Nelore; Bovine; ROS
• ISSN: 1469-8730
• DOI: 10.1017/S0967199421000502

The aim of this study was to evaluate follicular development, morphological integrity, and antioxidant potential of preantral ovarian follicles from Bos taurus indicus females grown in vitro with alpha-lipoic acid. Ovaries (n = 24) of Bos taurus indicus (n = 12) females were collected during slaughter and fragmented. A randomly obtained fragment from each pair of ovaries was fixed in Bouin (non-cultivated control; D0). These fragments were intended for classical histology (morphology and evaluation of follicular growth), and a fragment from each pair of ovaries was frozen at -80°C (non-cultivated control; D0), and assigned for analysis of oxidative stress [thiobarbituric acid reactive substances (TBARS), nitroblue tetrazolium (NBT), and ferric reducing antioxidant power (FRAP)]. The remaining fragments were cultured in vitro for 6 (D6) or 12 (D12) days, containing only minimum essential medium (MEM) or MEM supplemented with alpha-lipoic acid (50, 100, or 250 ng/ml), on an extracellular matrix of agarose gel, in an oven at 38.5ºC. Every 2 days, 100% of the culture medium was replaced. Supplementation with 100 ng/ml was effective for maintaining follicular integrity after 6 days of culture (primordial: 51.28%; development: 36.88%; P < 0.0001). There was no difference (P > 0.05) between treatments compared with the non-cultivated control treatment (D0), using the NBT and TBARS assays. Therefore, supplementation of the in vitro culture medium of bovine preantral ovarian follicles with a concentration of 100 ng/ml of alpha-lipoic acid at 6 days of culture was effective for maintaining follicular integrity and, after 6 days, maintaining stable levels of reactive oxygen species.