Further refinement of culture media is needed to improve the quality of embryos generated in vitro. Previous results from our laboratory demonstrated that uptake of nutrients by the embryo is significantly less than what is supplied in traditional culture media. Our objective was to determine the impact of reduced nutrient concentrations in culture medium on mouse embryo development, metabolism, and quality as a possible platform for next generation medium formulation. Concentrations of carbohydrates, amino acids, and vitamins could be reduced by 50% with no detrimental effects, but blastocyst development was impaired at 25% of standard nutrient provision (reduced nutrient medium; RN). Addition of pyruvate and L-lactate (+PL) to RN at 50% of standard concentrations restored blastocyst development, hatching, and cell number. In addition, blastocysts produced in RN + PL contained more ICM cells and ATP than blastocysts cultured in our control (100% nutrient) medium; however, metabolic activity was altered. Similarly, embryos produced in the RN medium with elevated (50% control) concentrations of pyruvate and lactate in the first step medium and EAA and Glu in the second step medium were competent to implant and develop into fetuses at a similar rate as embryos produced in the control medium. This novel approach to culture medium formulation could help define the optimal nutrient requirements of embryos in culture and provide a means of shifting metabolic activity towards the utilization of specific metabolic pathways that may be beneficial for embryo viability.

译文

需要进一步细化培养基,以提高体外产生的胚胎的质量。我们实验室的先前结果表明,胚胎对营养的吸收明显少于传统培养基中提供的营养。我们的目标是确定培养基中营养浓度降低对小鼠胚胎发育,代谢和质量的影响,作为下一代培养基配方的可能平台。50% 可以降低碳水化合物,氨基酸和维生素的浓度,而没有有害作用,但是在标准营养素供应 (营养培养基减少; RN) 的25% 下,囊胚发育受损。以50% 标准浓度向RN添加丙酮酸和L-乳酸 (PL) 可恢复胚泡发育,孵化和细胞数量。此外,在rn   +  PL中产生的囊胚比在我们的对照 (100% 营养) 培养基中培养的囊胚含有更多的ICM细胞和ATP; 然而,代谢活性发生了改变。类似地,在RN培养基中产生的胚胎具有升高 (50% 对照) 浓度的丙酮酸和乳酸在第一步培养基中以及在第二步培养基中产生的EAA和Glu的胚胎能够以与在对照培养基中产生的胚胎相似的速率植入和发育成胎儿。这种新颖的培养基配方方法可以帮助确定培养物中胚胎的最佳营养需求,并提供一种将代谢活性转移到利用可能有益于胚胎生存的特定代谢途径的手段。

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