新建辅助生殖实验室的鼠胚培养质控及鼠卵显微注射分析

Quality control of mouse embryo culture and microinjection analysis of mouse oocytes in a newly established assisted reproduction laboratory

目的利用小鼠体外受精法和体内受精法胚胎培养检测新建辅助生殖实验室的质控情况,分析比较常规人卵显微注射模式与鼠断尾精子显微注射模式的结局。方法利用昆明小鼠进行胚胎培养质控分析,按照受精方式不同分为体内受精组和体外受精组;又根据显微注射方式的不同,分为常规人卵显微注射模式(C-ICSI组)和鼠断尾精子显微注射模式(D-ICSI组),分析比较不同方式的鼠卵ICSI结局。结果体内受精组的受精率显著高于体外受精组(91.90%vs. 81.59%,P<0.05),两组的卵裂率(94.24%vs. 93.39%)、囊胚形成率(85.97%vs. 81.67%)比较无显著性差异(P>0.05)。人卵显微注射系统应用于鼠ICSI时卵子存活率有降低的趋势,但C-ICSI组与D-ICSI组的卵子存活率比较无显著性差异(29.49%vs. 30.52%)(P>0.05);D-ICSI组的受精率(56.72%vs. 42.53%)、卵裂率(71.05%vs. 43.24%)及囊胚形成率(61.11%vs. 31.25%)显著高于C-ICSI组(P<0.05)。结论昆明小鼠配子体外受精法和体内受精法胚胎培养的囊胚形成率均达到辅助生殖实验室的质控标准。人卵显微注射系统应用于鼠卵ICSI时,仅适用于卵子的显微注射操作练习。

Objective:To test the quality control status of the new assisted reproductive laboratory by in vitro fertilization and in vivo fertilization,and compare the results of the conventional human microinjection mode and the mouse sperm with broken tail microinjection mode.Methods:The quality control of embryo culture was tested by using Kunming mice.According to different fertilization methods,the mice were divided into in vivo fertilization group and in vitro fertilization group.According to different microinjection methods,it was divided into conventional human oocyte microinjection mode group(C-ICSI group)and mouse sperm with broken tail mouse microinjection mode group(D-ICSI group),and the ICSI outcomes in different modes were compared.Results:The fertilization rate of in vivo fertilization group was significantly higher than that of in vitro fertilization group(91.90%vs.81.59%,P<0.05),whereas the cleavage rate(94.24%vs.93.39%,P>0.05)and blastocyst formation rate(85.97%vs.81.67%,P>0.05)were not significantly different.The ovum survival rate was lower when human ICSI operating system was applied to the mice oocyte,but there was no significant difference in ovum survival between the C-ICSI group and the D-ICSI group(29.49%vs.30.52%,P>0.05).Compared with the C-ICSI group,the D-ICSI group had significantly higher fertilization rate(56.72%vs.42.53%),cleavage rate(71.05%vs.43.24%)and blastocyst formation rate(61.11%vs.31.25%)(P<0.05).Conclusions:The blastocyst formation rate of in vitro and in vivo fertilization of Kunming mice reaches the quality control standard of assisted reproduction laboratory.When the human ICSI operating system was applied to the mice oocyte,it was only suitable for the operation training of microinjection.