Lipid Enriched Reduced Nutrient Culture Medium Improves Bovine Blastocyst Formation.

IF 2.8 Q2 REPRODUCTIVE BIOLOGY Reproduction & fertility Pub Date : 2023-11-01 DOI:10.1530/RAF-23-0057
Rolando Pasquariello, Mingxiang Zhang, Jason R Herrick, Alison Ermisch, John Becker, William B Schoolcraft, Jennifer P Barfield, Ye Yuan, Rebecca L Krisher
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Abstract

The refinement of embryo culture media is essential in improving embryo viability and in vitro production efficiency. Our previous work demonstrated that the nutrients (carbohydrates, amino acids, and vitamins) in traditional culture media far exceed the need for an embryo and producing developmentally competent embryos in a reduced nutrient environment is feasible. Here, we aim to evaluate the impact of exogenous lipid and L-carnitine supplementation on bovine blastocyst development and refine our RN condition further. Zygotes were cultured in the control medium (100% nutrients) and reduced nutrient media containing 6.25% of the standard nutrient concentrations supplemented with L-carnitine and lipid free or lipid rich BSA. Increased blastocyst development was observed in the reduced nutrient lipid rich medium compared to the other two groups. However, in both reduced nutrient conditions, blastocyst cell numbers were lower than those obtained in the control condition. We then examined the expression level of 18 transcripts correlated with lipid metabolism, glucose metabolism, redox balance, and embryo quality, along with mitochondrial DNA copy numbers, ATP productions, and lipid profile. The results indicated lipid metabolism, embryo quality, and redox enzyme related genes were upregulated while glucose related gene was downregulated in embryos derived from reduced nutrient lipid rich condition Finally, we identified that the lipid rich BSA has enriched linoleic, stearic, oleic, palmitic, and alpha-linoleic fatty acids, a lipid profile that may contribute to the increased lipid metabolism and improved blastocyst development of the bovine embryos under the reduced nutrient condition.

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富脂减量培养基促进牛囊胚形成。
胚胎培养基的改良是提高胚胎存活率和体外生产效率的关键。我们之前的工作表明,传统培养基中的营养物质(碳水化合物、氨基酸和维生素)远远超过胚胎的需要,在营养减少的环境中产生发育能力强的胚胎是可行的。在这里,我们的目的是评估外源性脂质和左旋肉碱补充对牛囊胚发育的影响,并进一步完善我们的RN条件。受精卵分别在对照培养基(100%营养物)和含有6.25%标准营养物浓度的还原培养基中培养,培养基中添加左旋肉碱和无脂或富脂牛血清蛋白。与其他两组相比,在减少营养脂质的培养基中观察到囊胚发育增加。然而,在两种低营养条件下,囊胚细胞数量均低于对照组。然后,我们检测了与脂质代谢、葡萄糖代谢、氧化还原平衡、胚胎质量、线粒体DNA拷贝数、ATP产生和脂质谱相关的18个转录本的表达水平。结果表明,在营养脂质减少的条件下,胚胎的脂质代谢、胚胎质量和氧化还原酶相关基因上调,而葡萄糖相关基因下调。最后,我们发现富脂BSA富含亚油酸、硬脂酸、油酸、棕榈酸和α -亚油酸脂肪酸。在低营养条件下,脂质谱可能有助于提高牛胚胎的脂质代谢和改善囊胚发育。
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