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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、24 h)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1 h,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0.5 h、4 h、6 h,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 analyze the changes of amino acid metabolism in human early embryos after thawing, so as to find out the optimal timing of embryo thawing after thawing. Methods: From January 2013 to January 2014, 24 cases of embryos from 6 to 8 days after in vitro fertilization and embryo transfer in ovariectomized rats were collected and cultured for 2h After vitrification, frozen embryos were collected 2h before freezing and 15μL embryos cultured in each time after thawing. The culture medium without embryo culture in the same dish was used as the control group. The change of amino acid concentration of the samples at different time (0.5, 1, 2, 4, 6, 24 h) was detected by high performance liquid chromatography. Results: The levels of lysine, tryptophan, histidine and glutamine at different time points after thawing in the control group were significantly different from those before thawing (P <0.05), and the other 16 kinds of amino acids before and after thawing There was no significant difference between the two groups (P> 0.05). The amino acid concentrations in the embryo culture medium were significantly higher than those in the control group at 1 h after thawing (P <0.05), and the amino acid conversion was significantly higher than that before thawing (0.5 h, 4 h, 6 h, the difference was statistically significant (P <0.05). CONCLUSION: Early human embryos restored their metabolic status about 0.5 h after thawing, and began to perform amino acid metabolism. The level of amino acid metabolism was basically the same as that before freezing.