Culture of preantral ovarian follicles ofBos taurus indicuswith alpha-lipoic acid

• DOI: 10.1017/s0967199421000502
• Publication Date: 2021-08-25T10:10:17Z
• Authors: Larissa Zamparone...; Denis Vinicius Bo...; Camila Bizarro-Silva; Francieli Geslein...; Suellen Miguez Go...; Ana Carolina Ross...; Waldiceu A. Verri; Fábio Morotti; Marcelo Marcondes...
• Keywords: Tissue Culture Techniques; 03 medical and health sciences; 0302 clinical medicine; Ovarian Follicle; Thioctic Acid; Ovary; Animals; Cattle; Female; Antioxidants; Culture Media

SummaryThe aim of this study was to evaluate follicular development, morphological integrity, and antioxidant potential of preantral ovarian follicles fromBos taurus indicusfemales grownin vitrowith alpha-lipoic acid. Ovaries (n= 24) ofBos 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 culturedin vitrofor 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 thein vitroculture 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.