受精液、咖啡因、精液和抗菌素对猪卵母细胞的体外受精及早期胚胎发育的影响

探讨精液的不同处理、抗菌素、受精液种类和咖啡因对体外成熟猪卵母细胞的体外受精和早期胚胎发育的影响。结果表明：(1)受精液中抗菌素添加与否对体外成熟猪卵母细胞的体外受精无显著影响，而且不添加抗菌素的受精液也无严重微生物污染；(2)用猪冷冻精液与新鲜精液进行猪卵母细胞的体外受精对早期胚胎发育无显著影响；(3)在受精液mTBM中添加5mmol／L咖啡因对猪卵母细胞体外受精后的早期发育是最好的；(4)用mTBM进行猪卵母细胞体外受精的卵裂率明显高于用mBO进行体外受精的卵裂率，但其第8d囊胚率明显低千用mBO进行体外受精的，mTBM和mBO各有优势；(5)在洗精子液含有高浓度咖啡因(15mmol／L)的情况下，无咖啡因的受精液也可进行猪卵母细胞的体外受精并发育到囊胚阶段，在受精液中添加不同浓度(0～10mmol／L)的咖啡因对猪卵母细胞体外受精后的卵裂率和第8d囊胚率都无显著影响，受精液中咖啡因对猪卵母细胞体外受精后的第6d桑椹胚 囊胚率的影响因浓度而异．其中以添加5mmol／L为最佳，随着浓度上升效果开始下降。

Effects of semen,antibiotics,fertilization media and caffeine on fertilization and subsequent embryonic development of pig oocytes in vitro

In this study, the effects of boar semen, antibiotics, fertilization media and caffeine on in vitro fertilization and subsequent embryonic development of pig oocytes matured in vitro were investigated. The results of three experiments showed that: (1) supplementation of antibiotics in fertilization medium had no significant effect on in vitro fertilization of pig oocytes matured in vitro,and there was no microbiological contamination in the fertilization medium without antibiotics;(2) there was no significant effect on subsequent embryonic development of pig oocytes matured in vitro using frozen-thawed or fresh boar semen to fertilize in vitro; (3) the supplementation of 5 mmol/L caffeine in mTBM had the best effect on in vitro development of pig oocytes matured and fertilized in vitro; (4) the cleavage rate of pig oocytes inseminated in modified Tris buffered medium was significantly higher than that in modified BO medium, but the Day 8 blastocyst rate of pig oocytes inseminated in modified BO was significantly higher than that in modified Tris buffered medium, suggesting modified Tris buffered medium and modified BO have their own advantages; (5) In the presence of high concentration of caffeine (15 mmol/L) in sperm washing medium, pig oocytes could be fertilized in fertilization medium without caffeine and further develop to the blastocyst stage,and supplementation of different concentrations (0~10 mmol/L) of caffeine in fertilization medium had no significant difference on the cleavage and Day 8 blastocyst rates of pig oocytes matured and fertilized in vitro, and the effect of caffeine in fertilization medium on Day 6 morula+blastocyst rate of pig oocytes developed in vitro was concentration-dependent.