中文摘要：

猪卵母细胞体外受精（in vitro fertilization，IVF）技术为受精和胚胎早期发育机理等基础理论的研究及生产实践提供了重要的研究手段。虽然猪IVF的研究已经取得了一些进展，但其囊胚发育率仍然较低，亟需建立更为稳定、高效的体外受精方法。本试验通过比较精子浓度，精子获能处理，不同精卵共孵育培养体系以及在卵母细胞成熟过程中添加不同激素等影响猪卵母细胞体外受精胚胎发育能力的因素，以求找到最佳的猪卵母细胞体外受精体系。结果显示：在精子浓度为1×10^5-1×10^7／mL，5×10^6／mL的精子浓度显著提高体外受精效率（P〈0．05）；以茶碱、咖啡及咖啡因联合使用肝素作为获能物质处理精子，发现2．5mmol／L茶碱处理组体外受精效率显著提高（P〈0．05）；通过比较不同的受精体系，发现使用40个卵／500μL体系显著提高体外受精效率（P〈0．05）；在卵母细胞成熟液中添加不同激素组合，发现添加10IU／mL PMSG，10IU／mL HCG和2.5IU／mL FSH激素组合，体外受精效率显著提高（P〈0．05）。本试验结果表明，在M199中添加10IU／mL PMSG，10IU／mL HCG和2．5IU／mL FSH体外成熟培养卵母细胞，以5×10^6／mL精子浓度，2．5mmol／L茶碱作为获能物质处理精子，40个卵／500μL共孵育的IVF体系效果最佳，其卵裂率为（61．33±0．77）％，囊胚率为（28．33±1．08）％。本试验将为深入研究猪IVF提供理论参考。

英文摘要：

In vitro fertilization（IVF） of porcine oocytes provides an important method for the research of fertilization, early embryo development and production practice. Although the research of porcine IVF has made some progress,but its blastocyst development rate is still low. Therefore,it is urgent to establish more stable and efficient methods. The aim of the present study is to find the best IVF system in order to improve porcine IVF efficiency. Factors affecting the IVF including sperm concentration, capacitation treatment, co-incubation system, and hormone combination of in vitro maturation, were investigated in this study. Our results showed that sperm concentration at 5×10^6/mL significantly improved IVF efficiency in range from 1×10^5/mL to 1×10^7/mL（P〈0.05）. 2.5 mmo｜/L theophylline as the main capacitation component markedly enhanced IVF efficiency when compared with caffeine,or caffeine combined with heparin（P〈0.05）. 40 oocytes with 500μL co-incubation system obtained best IVF efficiency as compared other co-incubation system （P〈0.05）. Adding 10 IU/mL PMSG,10 IU/mL HCG and 2.5 IU/mL FSH into in vitro matura- tion medium acquired observable improvement of IVF efficiency as compared with other hormone combination addition（P〈0.05）. Our results indicated that the best cleavage rate（61.33±0.77）% and blastocyst rate（28. 33±1.08）% could be gained by the condition of adding 10 IU/mL PMSG, 10 IU/mL HCG and 2.5 IU/mL FSH into in vitro oocytes maturation medium, fertilized with 5×10^6/mL sperm concentration, capacitating sperm with 2.5 mmol/L theophylline, and co-incubating 40 oocytes at 500μL fertilization medium. These results provide a theoretical basis for the further study of porcine IVF.