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脂肪酸补充剂在玻璃化和温育过程中对小鼠、牛和人卵母细胞和胚胎发育能力的影响。

Effects of fatty acid supplementation during vitrification and warming on the developmental competence of mouse, bovine and human oocytes and embryos.

机构信息

Kato Ladies Clinic, 7-20-3 Nishishinjuku, Tokyo Shinjuku-ku 160-0023, Japan.

Kato Ladies Clinic, 7-20-3 Nishishinjuku, Tokyo Shinjuku-ku 160-0023, Japan.

出版信息

Reprod Biomed Online. 2021 Jul;43(1):14-25. doi: 10.1016/j.rbmo.2021.03.022. Epub 2021 Apr 20.

Abstract

RESEARCH QUESTION

Does fatty acid supplementation in vitrification and warming media influence developmental competence in oocytes after vitrification and warming?

DESIGN

Mouse oocytes and four-cell embryos were vitrified and warmed with solutions supplemented with fatty acid and cultured to the blastocyst stage. To study lipid metabolism after vitrification, quantitative real-time polymerase chain reaction was used to analyse the expression of genes related to beta oxidation in mouse embryos vitrified and warmed with or without fatty acids. The effects of fatty acid supplementation in the warming solutions on the developmental competence of bovine and human embryos were analysed. Blastocyst outgrowth assay was used to evaluate the potential of human blastocysts for adhesion to fibronectin.

RESULTS

The neutral lipid content of mouse oocytes in the fatty acid 1% supplementation group was significantly higher than in the fatty acid 0% group (P = 0.0032). The developmental rate to the blastocyst stage was significantly higher in the fatty acid 1% group than in the fatty acid 0% group in mice (P = 0.0345). Fatty acid supplementation in warming solution upregulated Acaa2 and Hadha in mouse embryos. Fatty acids significantly improved the developmental ability of bovine embryos to the blastocyst stage (P = 0.0048). Warming with 1% fatty acid supplementation significantly increased the proportion of human blastocysts with morphological grade A inner cell mass (P = 0.0074) and trophectoderm (P = 0.0323).

CONCLUSIONS

Fatty acid supplementation in the warming solutions improved the developmental competence of vitrified-warmed mouse oocytes by activating the beta-oxidation pathway. Fatty acid supplementation enhanced the developmental rate of bovine embryos to the blastocyst stage and improved morphological characteristics of human embryos vitrified at the cleavage stage.

摘要

研究问题

在玻璃化和温育介质中添加脂肪酸是否会影响玻璃化和温育后的卵母细胞的发育能力?

设计

使用添加脂肪酸的溶液对小鼠卵母细胞和四细胞胚胎进行玻璃化和温育,并培养至囊胚阶段。为了研究玻璃化后脂质代谢,使用定量实时聚合酶链反应分析了在有或没有脂肪酸的情况下玻璃化和温育的小鼠胚胎中与β氧化相关的基因的表达。分析了在温育溶液中添加脂肪酸对牛和人胚胎发育能力的影响。使用囊胚外伸测定法评估人囊胚对纤连蛋白黏附的潜力。

结果

添加脂肪酸 1%组的小鼠卵母细胞中性脂质含量明显高于添加脂肪酸 0%组(P=0.0032)。添加脂肪酸 1%组的卵母细胞发育为囊胚的比率明显高于添加脂肪酸 0%组(P=0.0345)。温育溶液中添加脂肪酸使小鼠胚胎中的 Acaa2 和 Hadha 上调。脂肪酸显著提高了牛胚胎发育为囊胚的能力(P=0.0048)。温育液中添加 1%脂肪酸显著增加了具有形态学 A 级内细胞团(P=0.0074)和滋养外胚层(P=0.0323)的人类囊胚的比例。

结论

温育溶液中添加脂肪酸通过激活β氧化途径提高了玻璃化温育卵母细胞的发育能力。脂肪酸补充剂提高了牛胚胎发育至囊胚阶段的速度,并改善了在卵裂期玻璃化的人类胚胎的形态特征。

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