Crocetin is an active constituent of saffron recently used as antioxidant for embryo culture. The aim of this study was to test the effect of crocetin added in the in vitro maturation (IVM) of prepubertal goat oocytes on the embryo development after in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and parthenogenetic activation (PA). Cumulus-oocyte complexes (COCs) were released from slaughterhouse ovaries of prepubertal goats and in vitro matured in supplemented TCM 199 medium during 24 h without (control group) and with crocetin. In Experiment 1, we evaluated the effect of the IVM supplementation with 0 μM (control), 0.5 μM, 1 μM and 2 μM of crocetin on the blastocyst development after IVF. No significant differences were obtained on blastocyst formation among groups (12, 7, 10, 11%; respectively). Although the blastocyst total cell number was higher in 1 μM crocetin group (150.7 cells) compared to the control (105.5), 0.5 μM (116.2) and 2 μM (93.7) crocetin groups, no significant differences were detected. In experiment 2, we assessed the effect of 1 μM crocetin supplementation in the IVM medium on the oocyte GSH level, ROS level and mitochondrial activity. ROS was significantly higher in the control than in the crocetin group (P < 0.05), but no differences in GSH level and mitochondrial activity were observed. In experiment 3, we evaluated the effect of 1 μM crocetin on the blastocyst development of oocytes after ICSI and PA. No statistical differences were found on blastocyst rate or cell number. However, compared with control, crocetin groups led to higher cleavage (59 vs. 67%, respectively, P = 0.09) and blastocyst rates (19 vs. 12%, respectively; P = 0.12) after ICSI. Although crocetin reduced ROS levels in prepubertal goat oocytes, it did not have a significant effect on oocyte embryo developmental competence.

中文翻译：

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在青春期前山羊卵母细胞的 IVM 培养基中添加藏红花素对体外受精、卵胞浆内单精子注射和孤雌生殖激活后囊胚结果的影响

Crocetin 是藏红花的活性成分，最近用作胚胎培养的抗氧化剂。本研究的目的是测试在青春期前山羊卵母细胞体外成熟（IVM）中添加藏红花素对体外受精（IVF）、卵胞浆内单精子注射（ICSI）和孤雌激活（PA）后胚胎发育的影响。卵丘-卵母细胞复合物 (COCs) 从青春期前山羊的屠宰场卵巢中释放出来，并在 24 小时内在补充 TCM 199 培养基中体外成熟，不含（对照组）和番红花素。在实验 1 中，我们评估了添加 0 μM（对照）、0.5 μM、1 μM 和 2 μM 藏红花素的 IVM 对 IVF 后囊胚发育的影响。各组之间的囊胚形成没有显着差异（分别为 12%、7%、10%、11%）。尽管与对照 (105.5)、0.5 μM (116.2) 和 2 μM (93.7) 藏红花素组相比，1 μM 藏红花素组（150.7 个细胞）的囊胚总细胞数更高，但未检测到显着差异。在实验 2 中，我们评估了在 IVM 培养基中添加 1 μM 藏红花素对卵母细胞 GSH 水平、ROS 水平和线粒体活性的影响。对照组的ROS显着高于藏红花素组（P < 0.05），但未观察到GSH水平和线粒体活性的差异。在实验 3 中，我们评估了 1 μM 藏红花素对 ICSI 和 PA 后卵母细胞胚泡发育的影响。在囊胚率或细胞数上没有发现统计学差异。然而，与对照组相比，藏红花素组导致更高的卵裂（分别为 59% 与 67%，P = 0.09）和囊胚率（19% 与 12%，分别; P = 0.12) 在 ICSI 之后。尽管藏红花素降低了青春期前山羊卵母细胞中的 ROS 水平，但它对卵母细胞胚胎发育能力没有显着影响。