To explore the development promoting mechanism of oviduct epithelial cells co-culture on the early embryos in mouse | 输卵管上皮细胞共培养促进小鼠早期胚胎发育机理的探索

Chinese. 为研究输卵管上皮细胞对小鼠早期胚胎发育质量及胚胎对共培养细胞活力和基因转录量的影响,探索输卵管上皮细胞共培养促进小鼠早期胚胎发育的机理,建立纯化的小鼠输卵管上皮细胞系并对合子进行共培养,用DCHFDA测定胚胎内部的H2O2含量,以MTT法测定输卵管上皮细胞活力,用RT-PCR检测CAT及GPX的转录状况。结果表明：1）共培养能够提高2-细胞早期、4-细胞期胚胎发育率（0.951 3% VS 0.903 3%,0.857 6% VS 0.700 5%,P〈0.05）和囊胚发育率（0.537 9% VS 0.573 3%,P 〉0.05）;2）2-细胞早期胚胎内部的H2O2含量显著降低（0.047 9 VS 0.068 2,P〈0.05）;3）Ⅰ和Ⅱ型阻滞胚胎的比例降低（0.158 3% VS 0.192 6%,0.213 3% VS 0.433 9%,P 〉0.05）,Ⅲ型阻滞胚胎的比例提高（0.621 6% VS 0.373 3%,P〈0.05）;4）共培养细胞活性上升（0.216 7 VS 0.195 7,P 〉0.05）;5）CAT及GPX的转录量增加（1.202 0 VS 0.984 9,1.310 1 VS 0.998 3,P 〉0.05）。结果显示输卵管上皮细胞共培养通过增强自身的抗氧化能力,利用细胞-细胞间的相互作用降低胚胎细胞内部的活性氧,提高胚胎质量,促进胚胎发育。

English. The objective of this study was to investigate the effect of oviduct epithelial cells on the development and quality of early embryos,as well as to study the impact of embryos on the activity and gene transcriptions of co-cultured oviduct epithelial cells,and explore the mechanisms on how oviduct epithelial cells improving the development of early embryo in mouse.Mouse oviduct epithelial cell lines were purified and used to co-culture with zygotes,H2O2 content in embryos was measured by DCHFDA,epithelial cell activity was measured by MTT,RT-PCR was applied to test the transcription of CAT and GPX gene.Results are as follows：1） co-culture can significantly improve the early 2-cell and 4-cell rates （0.951 3% VS 0.903 3%,0.857 6% VS 0.700 5%,P〈0.05） as well as blastocyst rate （0.537 9% VS 0.573 3%,P 〉0.05）;2） it can decrease the H2O2 content in early 2-cell embryos （0.047 9 VS 0.068 2,P〈0.05）;3） rates of type Ⅰ and Ⅱ arrested embryos tend to decrease （0.158 3% VS 0.192 6%,0.213 3% VS 0.433 9%,P 〉0.05）,rate of type Ⅲ arrested embryos was increased（0.621 6% VS 0.373 3%,P〈0.05）;4） cellular activity had improving trend when embryos developed to early 2-cell phase（0.216 7 VS 0.195 7,P 〉0.05）;5） the transcription of CAT and GPX were slightly increased（1.202 0 VS 0.984 9,1.310 1 VS 0.998 3,P 〉0.05）though not significant.Those results indicated that oviduct epithelial cell co-culture can improve the development of embryos by increasing anti-oxidative ability of themselves,then decreasing ROS content in embryos by cell-cell contact,therefore it can improve embryo quality and promote embryo development.