共培养对小鼠胚胎内部活性氧含量影响

文章旨在研究输卵管上皮细胞共培养对小鼠胚胎内部活性氧含量影响,探索共培养克服胚胎发育阻滞机理。用小鼠输卵管上皮细胞对合子进行共培养,DCFDA测定胚胎内部H2O2含量,以外源性H2O2对输卵管上皮细胞和胚胎进行阴性对照,用Hochest 33342和PI染色检测细胞凋亡情况,以MTT法测定输卵管上皮细胞活力。结果表明,共培养能显著提高2-细胞发育率、降低胚胎内部H2O2含量(P<0.05),提高4-细胞率及降低胚胎内部H2O2含量(P>0.05)。外源性H2O2能够使合子凋亡甚至死亡,导致共培养细胞活力下降(P<0.05),未能显著地促进胚胎发育(P>0.05)。结果表明共培养能够降低胚胎细胞内部活性氧,克服胚胎发育阻滞,促进胚胎发育。

Effect of co-culture on the reactive oxygen species content in mouse embryos

This experiment was conducted to study the effect of co-culture on the oxygen reactive content in mouse embryos,and explored the mechanisms of co-culture on overcoming the embryonic arrest.Mouse oviductal epithelial cells were used to co-cultured the zygotes,H2O2 content in embryos was measured by DCFDA,exogenous H2O2 was used as negative control to dispose epithelial cells and embryos,applied Hochest 33342 and PI staining to detect cellular apoptosis,MTT was used to measure epithelial cell activity.The results showed that co-culture can significantly improve 2-cell rate and decrease the H2O2 content in embryos(P<0.05),increase the rate of 4-cell embryos and reduce their cellular H2O2content(P>0.05),exogenous H2O2 can lead to embryonic apoptosis or death and decrease the cellular activity(P<0.05),therefore it cannot promote the development of embryos(P>0.05).Those results indicated that co-culture can decrease reactive oxygen species content in embryos,overcoming developmental arrest and promoting embryo development.