Correlation Between Glucose And Glutamine Consumption In Culture Medium And Bovine Embryonic Development Potential In Vitro

As livestock account for an increasing proportion in China’s breeding industry,in vitro embryo production technology is also developing steadily.Embryo quality assessment is one of the key steps in vitro embryo production,and this step is an important step in determining embryo quality and development potential.Current evaluation methods have certain limitations,either due to the subjective judgment of technical personnel,or the assessment method can cause harm to the embryo,affecting the quality of the embryo and the next step of development.In recent years,researchers have begun to focus on how to obtain more accurate embryo evaluation methods without harming embryos,and the evaluation through the analysis of material metabolic activities in the culture medium provides new ideas and new ideas for the assessment of embryonic development quality.Therefore,we wanted to detect the consumption of two important metabolic substances(i.e.,glucose and glutamine)in a single embryo culture medium,analyze their correlation with embryonic development potential,and lay the foundation for the subsequent establishment of harmless and highly sensitive embryonic development potential prediction and assessment techniques.In this experiment,bovine in vitro fertilization technology and single-drop singleembryo culture were combined to obtain corresponding embryo development data and collect culture medium for a specific time(i.e.,48 h after fertilization).In view of the current research gaps and lack of effective measurement techniques for glucose and glutamine in preimplantation embryonic development of bovines,we used a new multifunctional microoptical fiber biosensor that can realize ultra-sensitive and rapid detection of glucose and glutamine concentration in embryo culture medium to detect glucose and glutamine content in our collected embryo culture medium,and explored the correlation between glucose and glutamine consumption and embryonic development capacity.The results of this experiment showed that:(1)single drop single embryo culture using a round bottom 96-well plate can obtain a single blastocyst and a single embryo culture medium;(2)The utilization ability of embryos to glucose in the culture medium will gradually increase with embryonic development(P<0.05),and the content of glucose in the culture medium will gradually decrease with embryonic development(P<0.05)(the consumption of glucose by 2 cells,4 cells,8 cells and blastocysts is: 0.0722±0.0026mg/m L,0.0926±0.0110mg/m L,0.1674±0.0080 mg/m L,0.1986±0.0072mg/m L);The glucose consumed by embryonic development to blastocyst is in the range of 0.1907 mg/m L-0.2049mg/m L.(3)The consumption of glutamine in the culture medium by embryos will gradually increase with embryonic development(P<0.05),and the content of glucose in the culture medium will gradually decrease with embryonic development(P<0.05)(the consumption of glutamine by 2 cells,4 cells,8 cells and blastocysts is: 552.5495±21.3341μM,599.6057±14.0734μM,750.2591±20.8548μM,910.9662± 14.3090μM);The glutamine consumed by embryonic development to blastocyst is in the range of 894.5332μM-920.6706μM.The results showed that the consumption of glucose and glutamine in the culture medium correlated with the potential of embryonic development,and the consumption of glucose and glutamine gradually increased with embryonic development,and the consumption of glucose and glutamine was the highest when the embryo developed to blastocyst.The results of this study show that the method of using the micro-fiber biosensing platform to detect glucose and glutamine content in a single drop of single embryo culture medium is feasible and reliable,does not cause harm to embryos,and is fast and effective,which provides data support for the subsequent use of this method to evaluate embryonic development potential,and helps to realize the early screening and production utilization of high-quality bovine embryos.