时差成像培养系统对小鼠胚胎发育潜能及组蛋白修饰影响的研究  

Effect of time-lapse culture on the development and histone modification of mouse embryo

作　　者：李竞宇 李崇 黄国宁[1] Li Jingyu;Li Chong;Huang Guoning(Center for Reproductive Medicine,Women and Children's Hospital of Chongqing Medical University Chongqing Key Laboratory of Human Embryo Engineering,Chongqing 400013,China)

机构地区：[1]重庆医科大学附属妇女儿童医院生殖医学中心、人类胚胎工程重庆市重点实验室,重庆400013

出　　处：《中华生殖与避孕杂志》2023年第3期276-282,共7页Chinese Journal of Reproduction and Contraception

基　　金：重庆市科卫联合医学项目(2021MSXM072)。

摘　　要：目的评估时差成像培养系统(time-lapse imaging,TLI)对小鼠胚胎发育潜能及组蛋白表观修饰的影响。方法选择SPF级雌性ICR小鼠,促排卵后取成熟MⅡ卵子进行体外受精(in vitro fertilization,IVF),获取成功受精的合子,分别在TLI和常规培养体系(standard incubators,SI)中进行体外培养,记录两组胚胎的2-细胞率、4-细胞率、8-细胞率、桑葚胚率、囊胚率和孵出率。通过Hoechst、OCT4及CDX2抗体进行免疫荧光染色,分别统计囊胚细胞总数、内细胞团数和滋养外胚层细胞数。两组囊胚移植入第2.5天代孕母鼠子宫,统计植入率及活胎率。通过对组蛋白H3第4位赖氨酸三甲基化(H3K4me3)、组蛋白H3第9位赖氨酸二甲基化(H3K9me2)和组蛋白H3第9位赖氨酸乙酰化(H3K9ac)的免疫荧光染色,评估两组胚胎组蛋白表观修饰情况。结果TLI组和SI组的早期胚胎发育情况、内细胞团和滋养外胚层细胞数、植入率和活胎率比较差异均无统计学意义(均P>0.05)。两组囊胚的组蛋白H3K4me3、H3K9me2和H3K9ac的表达水平差异均无统计学意义(均P>0.05)。结论利用TLI体外培养不会影响胚胎的发育潜能以及组蛋白修饰。Objective To evaluate the effect of time-lapse imaging(TLI)culturing on the potential of mouse embryonic development and histone modification.Methods The matured MⅡoocytes were obtained after superovulation,using SPF female ICR mice.The successfully fertilized zygotes through in vitro fertilization(IVF)were assigned to culture either in TLI or standard incubator(SI).Immunofluorescent(IF)was performed to count the number of total cells,inner cell mass(ICM)and trophoblast cells(TE)in blastocysts,using the Hoechst,the antibody of OCT4 and CDX2,respectively.The rates of 2-cell,4-cell,8-cell,morula,blastocyst and blastocyst hatching were recorded.The blastocysts of the two groups were transferred into uteruses of day 2.5 pseudo pregnant mice,and the implantation rate and the live fetal rate were calculated.In addition,the levels of H3K4me3,H3K9me2,and H3K9ac were compared in blastocyst between the two groups using IF.Results No significant differences were found in the embryonic development,the cell number of ICM and TE,the implantation rate and the live fetal rate between TLI and SI groups(all P>0.05).Furthermore,there were no significant differences in the level of histone modification,including H3K4me3,H3K9me2,and H3K9ac(all P>0.05).Conclusion TLI culture does not have a significant adverse impact on the embryonic development and histone modification.

关 键 词：胚胎发育 时差成像培养系统 植入率 活胎率 组蛋白修饰 

分 类 号：R714.8[医药卫生—妇产科学]