人早期胚胎解冻后氨基酸代谢变化的临床分析

摘要

目的:分析人早期胚胎解冻后的氨基酸代谢变化,以明确冻融胚胎解冻后的最佳移植时机.方法:收集2013年1月-2014年1月本中心24例经体外受精-胚胎移植治疗患者取卵后第3日6～8细胞废弃胚胎,采用囊胚培养液微滴对其培养,培养2h后用玻璃化冷冻保存,于冷冻前2h、解冻后各时段分别收集15μL胚胎培养微滴,以同一培养皿中未进行胚胎培养的培养液为对照组.通过高效液相色谱法检测标本在不同时段(0.5、1、2、4、6、24h)的氨基酸浓度变化.结果:对照组解冻后不同时间点赖氨酸、色氨酸、组氨酸与谷氨酰胺浓度与与解冻前比较,差异具均有统计学意义(P＜0.05),其余16种氨基酸解冻前后浓度无明显变化,差异均无统计学意义(P＞0.05).胚胎培养液解冻后1h,胚胎培养液中的氨基酸浓度均明显高于对照组,差异具有统计学意义(P＜0.05),氨基转换量明显高于解冻前、解冻后0.5h、4h、6h,差异具有统计学意义(P＜0.05).结论:人早期胚胎在解冻后约0.5h就已开始恢复代谢状态,已开始进行氨基酸代谢,且氨基酸代谢水平与冷冻前基本相同.%Objective:To detection and analysis on changes in amino acid metabolism after human early embryos,in order to define the best opportunity for the transplantation of frozen thawed embryos.Methods:Selected 24 cases of third days after fertilization and embryo transfer treatment in patients with 6～8 cell abandoned embryos who treated in the center from January 2013 to January 2014,the blastocyst culture solution microdroplet was applied to culture such embryos,vitrification cryopreservation was taken after 2 hours of culture,15μ L of embryo culture droplets were respectively collected at time frames of 2 h before freezing and after thawing,embryos in the culture solution in the same petri dish bypassing embryo culture were selected into the control group.The changes in amino acid concentration were detected at different times via performance (0.5,1,2,4,6,24 h)by high performance liquid chromatography.Results:In the control group,the concentration of lysine,tryptophan,histidine and glutamine after thawing at the different time points were statistically significant compared with before thawing (P＜0.05),the remaining 16 amino acids before and after thawing concentration had no significant changes,the difference were not statistically significant (P＞0.05),1 h after thawing,the concentrations of amino acids in embryo culture medium were significantly higher than those in the control group,the differences were statistically significant (P＜0.05),thawing amino conversion were significantly higher than that of before thawing,0.5 h,4 h,and 6 h,the differences were statistically significant (P＜0.05).Conclusion:Human early embryos have begun to recover about 0.5 h after thawing,amino acid metabolism has been started,and its metabolism of amino acid is basic same to that before thawing.