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.

本研究的目的是评估用 α-硫辛酸在体外生长的金牛雌性卵泡的卵泡发育、形态完整性和抗氧化能力。Bos taurus indicus ( n = 24) 的卵巢 ( n = 24)= 12) 雌性在屠宰期间被收集并分割。从每对卵巢中随机获得的片段固定在 Bouin 中（非培养对照；D0）。这些片段用于经典组织学（卵泡生长的形态学和评估），每对卵巢的片段在 -80°C 下冷冻（非培养对照；D0），并指定用于分析氧化应激 [硫代巴比妥酸活性物质 (TBARS)、硝基蓝四唑 (NBT) 和铁还原抗氧化能力 (FRAP)]。剩余的片段在体外培养6 (D6) 或 12 (D12) 天，在琼脂糖凝胶的细胞外基质中，仅含有最低限度的必需培养基 (MEM) 或补充有 α-硫辛酸（50、100 或 250 ng/ml）的 MEM烤箱温度 38.5ºC。每 2 天更换 100% 的培养基。培养 6 天后，补充 100 ng/ml 可有效维持卵泡完整性（原始：51.28%；发育：36.88%；P < 0.0001）。使用 NBT 和 TBARS 测定，处理与非栽培对照处理 (D0) 之间没有差异 ( P > 0.05)。因此，体外补充牛窦前卵泡培养基在培养 6 天时，α-硫辛酸浓度为 100 ng/ml，可有效维持卵泡完整性，并在 6 天后维持稳定的活性氧水平。