A novel culture medium with reduced nutrient concentrations supports the development and viability of mouse embryos

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 im...

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Bibliographic Details
Published in:	Scientific reports 2020-06, Vol.10 (1), p.9263, Article 9263
Main Authors:	Ermisch, Alison F, Herrick, Jason R, Pasquariello, Rolando, Dyer, McKenna C, Lyons, Sarah M, Broeckling, Corey D, Rajput, Sandeep K, Schoolcraft, William B, Krisher, Rebecca L
Format:	Article
Language:	eng
Subjects:	Amino Acids, Essential - pharmacology Animals Blastocyst - cytology Blastocyst - drug effects Blastocyst - physiology Blastocysts Carbohydrates Cell culture Cell number Culture media Culture Media - chemistry Culture Media - pharmacology Dipeptides - pharmacology Edetic Acid - pharmacology Embryo Culture Techniques - methods Embryo Transfer Embryos Female Fertilization in Vitro Fetuses Gene Expression Regulation, Developmental - drug effects Glucose - pharmacology Hatching Lactic acid Lactic Acid - pharmacology Metabolic pathways Metabolism Mice Nutrient concentrations Nutrient requirements Nutrients Pyruvic acid Pyruvic Acid - pharmacology Vitamins
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Summary:	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.
ISSN:	2045-2322 2045-2322