【摘要】 本试验探讨了不同辅助激活方法(Calciumionophore A23187激活、Calciumionophore A23187+6-DMAP联合激活和电激活)、不同精子预处理方法(液氮冻融处理和0.1%Triton X-100处理)和在添加半胱氨酸的胚胎培养液中培养不同时间(0 h、4 h、12 h和168 h)对猪卵母细胞内单精子注射(ICSI)胚胎体外发育的影响。结果显示:与无辅助激活相比,A23187+6-DMAP联合激活和电激活均能显著提高ICSI卵母细胞的激活率、卵裂率和囊胚率(P<0.05),A23187+6-DMAP联合激活能显著提高ICSI卵母细胞的受精率(P<0.05)。液氮冻融精子组ICSI卵母细胞的雄原核形成率显著高于活精子组(P<0.05)。在添加半胱氨酸的胚胎培养液中培养4 h的ICSI卵母细胞受精率、雄原核形成率和囊胚率显著高于0 h组(P<0.05)。以上结果表明,猪卵母细胞在ICSI后需要辅助激活来启动胚胎顺利发育,A23187+6-DMAP激活效果较好。液氮冻融精子可以促进ICSI后雄原核的形成。半胱氨酸处理4 h对猪ICSI卵母细胞受精和发育均有促进作用。更多还原

【Abstract】 The objective of this study was to investigate the effects of various methods of oocyte activation(Calcium ionophore A23187,Calcium ionophore A23187+6-DMAP and DC),sperm pretreatment(sperm rendered immotile by one-time freezing and thawing without cryoprotectant and pre-incubating with 0.1% Triton X-100) and cysteine treatment(0 h,4 h,12 h and 168 h) on development of porcine embryos derived from intracytoplasmic sperm injection(ICSI) of in vitro matured oocytes.The results showed that: 1) The oocyte activation,cleavage and blastocyst rates of A23187+6-DMAP and DC groups significantly increased compared to no additional activation group(P<0.05).The fertilization rates of A23187+6-DMAP group significantly increased compared to no additional activation group(P<0.05).2) The male pronucleus rates in frozen/thawed sperm group were significantly higher than motile sperm group(P<0.05).3) Fertilization,male pronucleus and blastocyst rates significantly increased when presumed zygotes after ICSI were cultured with cysteine for 4 h compared to 0 h group(P<0.05).These results indicated that additional activation of the oocytes is necessary for the porcine ICSI,and A23187+6-DMAP is a reasonable choice;Pretreatment of sperm by freezing and thawing without cryoprotectant can promote the male pronucleus formation of the presumed zygotes after ICSI;Culture of presumed zygotes after ICSI with cysteine for 4 h can promote fertilization and development.更多还原