血清及山羊卵丘细胞共培养对猪胚胎体外发育的促进作用

Effects of serum and coculture with goat cumulus cells on in vitro development of porcine embryos

为进一步完善猪胚胎体外发育系统,以孤雌激活胚胎为研究对象,依次对胚胎培养液种类、培养期间血清添加方式、卵丘细胞共培养体系进行筛选优化。结果表明,与常规的PZM3培养液相比,CR1aa并不适合猪胚胎的体外发育;猪胚胎在PZM3中培养4d后添加10%胎牛血清培养至第6天,体外发育能力得到显著改善,卵裂率、囊胚率、囊胚孵化率、囊胚细胞数分别达到95.1%、56.5%、27.3%、79.3;在此基础上,利用山羊卵丘细胞与猪胚胎进行共培养,将囊胚孵化率进一步提高至36.6%,初步建立了一套高效稳定的猪胚胎体外发育系统。

To further improve the in vitro development systems of porcine embryos,we optimized the types of embryo culture media,the methods of serum treatment during embryo culture,the effects of coculture with cumulus cells on embryos development in vitro,using porcine parthenogenetic embryos as a model.The results demonstrated that porcine zygote medium-3(PZM3)was the suitable medium for in vitro culture of porcine embryos compared with Charles Rosenkrans 1(CR1aa).Cultured in PZM3from Day 0to 4and then added 10%fetal bovine serum from Day 4to 6,porcine embryos showed the higher developmental capacity,with the cleavage rates,blastocyst rates,hatched blastocyst rates and cell number of blastocysts of 95.1%,56.5%,27.3%and 79.3respectively.Based on the above,we cocultured porcine embryos with goat cumulus cells and further improved the hatched blastocyst rates to 36.6%,suggesting an efficient and stable in vitro development systems of porcine embryos was established preliminarily.