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在卵母细胞成熟过程中添加生长因子和抗氧化剂优化猪的体外胚胎生产。

Optimizing swine in vitro embryo production with growth factor and antioxidant supplementation during oocyte maturation.

机构信息

Department of Animal Science, McGill University, Sainte-Anne-de-Bellevue, Quebec, Canada.

Department of Animal Science, McGill University, Sainte-Anne-de-Bellevue, Quebec, Canada.

出版信息

Theriogenology. 2022 Dec;194:133-143. doi: 10.1016/j.theriogenology.2022.10.005. Epub 2022 Oct 8.

Abstract

Porcine in vitro fertilization often results in low embryo development rates compared to other livestock species, which is often associated with either a low fertilization rate or high incidence of polyspermy. Since the quality of oocyte maturation is known to play a significant role in oocyte competence, we investigated the impact of supplementing in vitro maturation (IVM) medium containing porcine follicular fluid (pFF) with the growth factors FGF2, LIF and IGF1 (FLI), along with different combinations of cysteine, melatonin and ITS, on cumulus cell expansion, oocyte meiotic maturation, fertilization outcome, embryo development and blastocyst cell numbers. Maturation medium containing pFF yielded the greatest cumulus expansion. Compared to pFF and FLI individually, using pFF and FLI together resulted in the best embryo development rates over total oocyte placed in IVF (12.5% vs. 15.0% vs. 26.6%, respectively). Supplementation of IVM medium containing pFF and FLI with either cysteine, melatonin or insulin-transferrin-selenium, revealed that cysteine was essential to improve embryo development, while melatonin and ITS had a limited impact on improving blastocyst rates. Finally, we observed that pig oocytes matured in medium supplemented with pFF, FLI, cysteine and melatonin had a high proportion of monospermic zygotes (68.2%) and low proportion of polyspermic zygotes (15.9%) following IVF and yielded superior cleavage (78.2%) and blastocyst (32.0%) rates.

摘要

猪的体外受精常导致胚胎发育率低,与其他家畜相比,这通常与受精率低或多精受精率高有关。由于卵母细胞成熟质量已知在卵母细胞功能中起重要作用,我们研究了在含有猪卵泡液(pFF)的体外成熟(IVM)培养基中添加生长因子 FGF2、LIF 和 IGF1(FLI),以及不同组合的半胱氨酸、褪黑素和 ITS 对卵丘细胞扩展、卵母细胞减数分裂成熟、受精结果、胚胎发育和囊胚细胞数的影响。含有 pFF 的成熟培养基产生了最大的卵丘扩展。与 pFF 和 FLI 单独使用相比,一起使用 pFF 和 FLI 可使总卵母细胞在 IVF 中获得最佳的胚胎发育率(分别为 12.5%、15.0%和 26.6%)。在含有 pFF 和 FLI 的 IVM 培养基中添加半胱氨酸、褪黑素或胰岛素-转铁蛋白-硒,结果表明,半胱氨酸对于提高胚胎发育至关重要,而褪黑素和 ITS 对提高囊胚率的影响有限。最后,我们观察到在添加了 pFF、FLI、半胱氨酸和褪黑素的培养基中成熟的猪卵母细胞在 IVF 后具有高比例的单精受精合子(68.2%)和低比例的多精受精合子(15.9%),并产生了较高的卵裂(78.2%)和囊胚(32.0%)率。

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