序贯共培养体系对小鼠早期胚胎体外发育效果的影响

将体外分离培养经超排处理的新西兰大白兔输卵管上皮及子宫内膜细胞接种于4孔板培养。处理1以CZB为培养液;处理2以CZB加输卵管上皮细胞为培养体系;处理3以CZB加子宫内膜细胞为培养体系;处理4以CZB为培养液,胚胎培养的前24h在输卵管上皮细胞共培养体系中,之后则转入子宫内膜细胞共培养体系中培养(处理4为序贯共培养体系)。将2-细胞期小鼠胚胎随机分配给4个处理,统计各处理组小鼠胚胎的各期发育率及发育质量。结果表明,序贯共培养体系胚胎的囊胚体外发育率显著高于其他共培养组(P<0.05);囊胚孵化率及孵化囊胚的贴附率显著高于其他共培养组(P<0.05);囊胚细胞总数各共培养组显著高于单独培养液组(P<0.05)。可见序贯共培养体系较单一体细胞共培养体系能更好地模拟体内环境,提高早期胚胎的发育率和发育质量。

Effect of sequential co-culture system on the development and quality of marine embryos in vitro

This study is to establish an effective co-culture system to improve the development rate and quality of early embryos in vitro.The oviduct epithelial cells and endometrial cells were isolated and cultured in vitro,then inoculated into 4-well plate.The 2-cell embryos of murine were cultured in single CZB media in treatment 1,co-cultured with oviduct epithelial cells in CZB in treatment 2,with endometrial cells in CZB in treatment 3 and with oviduct epithelial cells in CZB in the first 24 hours,and then transferred into the endometrial cells co-culture system to fulfill the culture in vitro in treatment 4.The results showed that more embryos in the sequential co-culture system reached the blastocyst stage than that in other co-culture system (P<0.05);Sequential co-culture system could increase the hatching rate of the blastocysts and the attachment rate of hatched blastocysts compared to the other co-culture system (P<0.05);All co-culture system could increase the cell number of the blastocyst compared with the single medium culture system (P<0.05).This study demonstrates that the sequential co-culture system can better simulate the natural developing environment of embryos in vitro and improve the development rate and quality of early embryos.