【摘要】 通过优化猪卵母细胞体外成熟、体外受精和胚胎体外发育体系,以进一步提高体外胚胎的生产效率和质量。研究了激素存在时间、不同激素和不同血清对猪卵母细胞体外成熟的影响;共培养体系、精卵作用时间、去除卵丘细胞的方法对猪体外受精及早期胚胎发育的影响。猪卵母细胞IVM培养48h,前24h内加入PMSG、hCG,后24h将其去除,卵母细胞总成熟率为79.54%;培养液添加15%FCS或15%NCS,卵母细胞成熟率分别为79.48%和74.81%;PMSG、HCG和E2配合使用后卵母细胞成熟率为81.42%。在IVF前用吹打法获得的卵裂率、桑椹胚率分别为37.89%和8.54%,精卵共孵育6h或8h的卵裂率(40.52%,37.24%)、桑椹胚率(8.42%,7.85%),以及用输卵管上皮细胞共培养所获得的卵裂率(40.84%)、桑椹胚率(9.53%)均显著高于其它各组。更多还原

【Abstract】 To optimize the systems of IVM,IVF of oocytes and IVC of embryos on porcine,and further improve the efficiency and quality of porcine embryos in vitro.We studied some factors affect on oocytes in vitro maturation,the factors include different time of PMSG and hCG added,as well as different type of serums and hormones;We also studied some factors affect on fertilization,this factors include different co-cultured system,the removal of cumulus,and the time of sperm-egg incubation.The results show that when oocytes cultured for 48 hours in vitro,the oocytes maturation rate got 79.54% by removing PMSG and hCG from nutrient medium after 24 hours;got 79.48% and 74.81% by using culture medium containing 15%FCS or 15%NCS;got 81.42% by adding PMSG,hCG and E2,respectively.The cleavage rate was 37.89% and morula rate was 8.54% by manual removal of cumulus in before IVF.the better cleavage rate was 40.84% and the morula rate was 9.53% by using co-cultured system with POEC(porcine oviduct epithelial cell monolayer)in vitro fertilization.Time of fertilization had important effect on cleavage and embryo developing capacity,which is either the time is 6 hours or 8 hours has no significant difference on embryo development,but 6 hours and 8 hours are better than the others.更多还原