新建辅助生殖实验室体外受精胚胎培养技术的小鼠实验评价  

Assessment on in-vitro fertilized embryo culture capability in new ART laboratory by using mice oocytes

作　　者：桂俊豪 杨楠[2] 常艳艳 苏雪梅[2] GUI junhao;YANG Nan;CHANG Yanyan;SU Xuemei(Reprodutive Medicine Center of Chengdu Fifth People′s Hospital,Chengdu,Sichuan 611130)

机构地区：[1]成都市第五人民医院生殖医学中心,611130 [2]新疆军区总医院生殖医学中心,乌鲁木齐830000

出　　处：《中国生育健康杂志》2023年第3期261-264,共4页Chinese Journal of Reproductive Health

摘　　要：目的评价新建胚胎实验室培养环境及培养体系的安全性和可靠性及操作者的技术水平。方法将体重25~35 g成年昆明雌性小鼠40只随机分为体内受精组与体外受精组,两组均采用孕马血清促性腺激素(PMSG)和HCG 10 IU/只为腹腔内满程进针注射促排卵。体内受精组:将注射HCG后的雌鼠与体重42~50 g成年雄鼠按1:1比例合笼后处死母鼠,收集受精卵进行体外培养;体外受精组:分别取卵、取精后开展常规体外受精,于受精后20 h观察2-细胞,30~40 h观察4-细胞,60~70 h观察8-细胞及桑葚胚,90 h后观察囊胚的数量。比较体内受精组和体外受精组内、组间卵裂率和囊胚形成率。结果共培养观察7个批次,两组各20个周期,体内受精和体外受精组分别获得受精卵752和805个;两组的卵裂率分别为94.1%和89.4%;囊胚形成率分别为92.1%和90.3%。体内受精组的卵裂率高于体外受精组(P<0.05),两组的囊胚形成率差异未见统计学显著性。结论该新建辅助生殖实验室体外受精胚胎培养体系的囊胚形成率符合质量要求,实验室的胚胎培养技术达到了质控标准。Objective To assess the reliability of our newly constructed laboratory for in vitro fertilization as well as embryo culture,and the capability of operator′s technical manipulation of.Methods A total of 40 mice Kunming breed mice(25~35 g)were randomly divided into in vitro fertilization(IVF)group and in vivo fertilization group,followed by superovulating with pregnant mare serum gonadotropin(PMSG)and with 10 IU human chorionic gonadotropin(hCG)per female mice by intraperitoneal injection.Especially,the needle of syringes were fully pierced into the abdomen of female mice to avoid leakage or residual of PMSG and HCG.Additionally,male mice(42-50 g)were used as sperm donors.The female mice of in vivo group were housed together overnight with male mice according to the gender ratio of 1:1,then the fertilized female mice were sacrificed by cervical dislocation and oviducts were removed for fertilized oocytes retrieval.As for in vitro group,the routine IVF procedure was performed after oocyte and sperm collection.During embryo culture,2-cell,4-cell or 8-cell embryos,morula and blastocyst were recorded respectively at 20h,30-40h,60-70h or 90h after in vivo or in vitro fertilization.The cleavage rate and blastocyst formation rate of two groups were recorded and compared.Results in vivo or in vitro fertilization experiments were performed for seven batches and a total of 20 cycles were finished in each group,and 752 and 805 fertilized oocytes were obtained from in-vivo and in-vitro groups,respectively.Statistical analysis indicated that the cleavage rate of in vivo fertilization group was 94.1%,significantly higher than 89.4%in-vitro fertilization group(P<0.05).Additionally,the blastocyst formation rates of in-vivo and in-vitro groups were 92.1%and 90.3%,respectively(P>0.05).Conclusion The blastocyst formation rates of in-vitro and in-vivio fertilization and the embryo culture procedures of the newly established reproductive medicine laboratory have reached the quality control standard.

关 键 词：鼠胚实验 体外受精 体内受精 胚胎培养室 质量控制 

分 类 号：R71[医药卫生—妇产科学]