新建体外受精实验室的鼠胚实验质控和氧气浓度对胚胎体外发育的影响

Application of mouse embryo experiment for quality control of IVF laboratory and the effect of oxygen concentration on embryo development in vitro

目的采用鼠胚实验(MEA)检测新建体外受精(IVF)实验室的培养环境和培养体系是否符合辅助生殖技术人类胚胎体外培养的要求,并探讨不同培养环境(O2浓度)对小鼠早期胚胎体外发育的影响。方法利用ICR小鼠进行体内受精和IVF实验,观察不同受精方式来源的早期胚胎的受精及发育情况,并比较不同O2浓度下体内受精/IVF早期胚胎的受精率、卵裂率和囊胚形成率。结果体内受精组共获卵1387个,受精率、卵裂率和囊胚形成率分别为86.37%、94.74%和91.72%;IVF组共获卵1345个,受精率、卵裂率和囊胚形成率分别为80.67%、94.75%和91.05%。体内受精组的受精率明显高于IVF组(P<0.05),但两组卵裂率和囊胚形成率比较差异均无统计学意义(均P>0.05)。体内受精组中,二气培养(三气培养)的受精率、卵裂率和囊胚形成率分别为87.04%、93.09%和88.19%(85.79%、96.21%和94.75%),二气培养与三气培养受精率比较差异无统计学意义(P>0.05),但二气培养卵裂率和囊胚形成率均低于三气培养(均P<0.05)。IVF组的二气和三气培养环境下的鼠胚发育情况与体内受精组一致。结论MEA IVF和体内受精的囊胚形成率均达到了IVF实验室的质控标准。建议IVF实验室进行早期胚胎体外培养时选用三气培养,其可有效减少活性氧对早期胚胎的氧化应激作用,并可提高其发育潜能。

Objective To apply mouse embryo assay(MEA)for quality control of in vitro fertilization(IVF)laboratory and to investigate the effects of oxygen concentration on the embryo development in vitro.Methods ICR mouse were used for in vivo and IVF experiments.The fertilization and development of early embryos from different fertilization methods were observed.The effects of O2 concentration on fertilization rate,cleavage rate and blastocyst formation rate of mouse early embryos were investigated.Results In the in vivo fertilization experiments,1387 eggs were collected and the fertilization rate,cleavage rate and blastocyst formation rate were 86.37%,94.74%and 91.72%,respectively.In the IVF experiments,1345 oocytes were retrieved,and the fertilization rate,cleavage rate and blastocyst formation rate were 80.67%,94.75%and 91.05%.The fertilization rate of in vivo fertilization group was significantly higher than that of IVF group(P<0.05),but there was no significant difference in cleavage rate and blastocyst formation rate between two groups(both P>0.05).In the in vivo fertilization experiments,the fertilization rate,cleavage rate and blastocyst formation rate of two-gas culture and tri-gas culture were 87.04%,93.09%,88.19%and 85.79%,96.21%,94.75%,respectively.There was no significant difference regarding fertilization rate between two-gas culture and tri-gas culture(P>0.05),but the cleavage rate and blastocyst formation rate of two-gas culture were significantly lower than those of tri-gas culture(both P<0.05).In the IVF experiments,the development of mouse embryos in both two-gas and tri-gas culture environment was comparable with that in the in vivo fertilization.Conclusion The blastocyst formation rates of in vivo and IVF meet the quality control standard of IVF laboratory.The use of tri-gas culture for in vitro culture of early embryos in IVF laboratory is recommended,which can effectively reduce the oxidative stress brought by reactive oxygen species and can improve developmental potential of early embryo.