Yunnan Animal Science and Veterinary Institute, Jindian, Panlong County, Kunming City, Yunnan province, China; Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Jindian, Panlong County, Kunming City, Yunnan province, China.
Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences (CAAS), No.2 Yuanmingyuan Western Road, Haidian District, Beijing, 100193, China.
Cryobiology. 2020 Apr;93:84-90. doi: 10.1016/j.cryobiol.2020.02.004. Epub 2020 Feb 7.
The purpose of this present study is to assess if addition of the synthetic polymers in maturation medium can influence cryotolerance and subsequently embryonic development of mammalian oocytes. We examined the roles of two polymers, including polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP), on in vitro maturation (IVM), embryonic developmental capacity, and cryotolerance of goat oocytes. The present study includes two parts. At first, goat cumulus-oocyte complexes (COCs) were matured in a medium supplemented with 10% fetal bovine serum (FBS), 3 mg/ml PVP, or 1 mg/ml PVA, respectively. Data of oocyte with first polar body, cleavage, and blastocyst following parthenogenetic activation (PA) were recorded. Secondly, after maturation in the above medium, oocytes were vitrified using the Cryotop technique and then the morphology, cleavage and blastocyst formation of vitrified oocytes have been checked. The results demonstrated that the adding of PVP or PVA in maturation medium can't affect IVM of goat oocytes in comparison with FBS, as concern cumulus cell expansion, first polar body formation, and embryonic development. Additionally, without plunging into liquid nitrogen, only exposure to the vitrification and warming solutions cannot also influence the quality of oocytes, in terms of morphology, cleavage, and blastocyst formation. However, after IVM with synthetic polymers and vitrification, the ratio of oocytes with standard morphology in PVP or PVA group was only 59.47% ± 3.56% or 54.86% ± 5.19%, respectively, and was significantly less than that in the FBS group (89.37% ± 4.52%, P < 0.05). Furthermore, the cleavage ratio of oocytes in PVP or PVA group was 37.41% ± 4.17% or 27.71% ± 3.91% and was considerably less than that in the FBS group (64.97% ± 4.69%, P < 0.05). In addition, the cleavage ratio in PVP group was statistically higher than that in PVA group (P < 0.05). In terms of blastocyst development, a significant difference was observed between the synthetic polymer group and the FBS group (24.96% ± 3.62%, P < 0.05). However, the blastocyst ratio in the PVA group (7.51% ± 1.68%) was statistically less than the PVP groups (13.20% ± 4.59%, P < 0.05) and the FBS group (P < 0.05). In conclusion, two potential serum replacements, either PVP or PVA, can support IVM and embryonic development of goat oocytes at the concentration used in this study. But IVM with synthetic polymers supplemented to maturation medium may reduce the cryotolerance of oocytes. Additionally, the supportive function of PVP on embryonic development of vitrified oocytes might be better than that of PVA.
本研究旨在评估在成熟培养基中添加合成聚合物是否会影响哺乳动物卵母细胞的冷冻耐受性,进而影响胚胎发育。我们研究了两种聚合物,包括聚乙烯醇(PVA)和聚乙烯吡咯烷酮(PVP),对山羊卵母细胞体外成熟(IVM)、胚胎发育能力和冷冻耐受性的影响。本研究包括两个部分。首先,山羊卵丘-卵母细胞复合物(COCs)分别在添加 10%胎牛血清(FBS)、3mg/ml PVP 或 1mg/ml PVA 的培养基中进行成熟。记录了经孤雌激活(PA)后第一极体、卵裂和囊胚的数据。其次,在上述培养基中成熟后,使用 Cryotop 技术对卵母细胞进行玻璃化冷冻,然后检查玻璃化冷冻卵母细胞的形态、卵裂和囊胚形成情况。结果表明,与 FBS 相比,添加 PVP 或 PVA 对山羊卵母细胞的 IVM 没有影响,与卵丘细胞扩张、第一极体形成和胚胎发育有关。此外,在不浸入液氮的情况下,仅暴露于玻璃化和预热溶液中也不会影响卵母细胞的质量,无论是形态、卵裂还是囊胚形成。然而,在合成聚合物和玻璃化冷冻后的 IVM 中,PVP 或 PVA 组中具有标准形态的卵母细胞比例仅为 59.47%±3.56%或 54.86%±5.19%,明显低于 FBS 组(89.37%±4.52%,P<0.05)。此外,PVP 或 PVA 组的卵裂率为 37.41%±4.17%或 27.71%±3.91%,明显低于 FBS 组(64.97%±4.69%,P<0.05)。此外,PVP 组的卵裂率明显高于 PVA 组(P<0.05)。在囊胚发育方面,合成聚合物组与 FBS 组之间存在显著差异(24.96%±3.62%,P<0.05)。然而,PVA 组的囊胚比例(7.51%±1.68%)明显低于 PVP 组(13.20%±4.59%,P<0.05)和 FBS 组(P<0.05)。总之,在本研究中使用的浓度下,两种潜在的血清替代品,PVP 或 PVA,都可以支持山羊卵母细胞的 IVM 和胚胎发育。但是,在成熟培养基中添加合成聚合物可能会降低卵母细胞的冷冻耐受性。此外,PVP 对玻璃化冷冻卵母细胞胚胎发育的支持作用可能优于 PVA。