中文摘要：

本试验探讨了不同辅助激活方法（Calcium ionophore A23187激活、Calcium ionophore 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（0h、4h、12h和168h）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4h的ICSI卵母细胞受精率、雄原核形成率和囊胚率显著高于0h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4h对猪ICSI卵母细胞受精和发育均有促进作用。

英文摘要：

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.