Abstract: In an attempt to improve the in vitro fertilization(IVF)efficacy of bovine sexing embryo,a set of key technology for improving IVF efficacy of bovine sexing semen was developed.Firstly,the effect of three in vitro capacitation methods of bovine sexing frozen semen and antioxidant additive addition on IVF efficacy of bovine sexing semen was statistically evaluated;secondly,the impact of partial removal of cumulus cell from oocyte matured in vitro for 24 h on IVF efficacy of bovine sexing embryo was assessed.The results showed that while the cleavage rate and blastocyst rate of IVF produced zygotes of one-time centrifugation in vitro capacitation method[(68.3±3.7)%,(30.9±2.3)%]had no significant differences with sperm floating capacitation method[(67.8±2.6)%,(29.7±3.7)%](P>0.05),they were significantly higher than those of two-time centrifugation in vitro capacitation method[(56.1±4.1)%,(21.6±4.6)%](P<0.05);the treatment with addition of 1.0%ascorbic acid in capacitation liquid and IVF liquid had significantly higher cleavage rate[(70.2±3.2)%]and blastocyst rate[(35.0±4.7)%]of IVF produced zygotes compared to the treatment without additive[(60.0±4.5)%,(28.3±3.6)%](P<0.05);the cleavage rate and blastocyst rate of IVF produced zygotes in the treatment that the surrounding cumulus cell was partially removed from the oocyte prior to IVF[(70.1±4.6)%,(35.5±3.7)%]were found significantly higher compared to the treatment without cumulus cell removal[(52.9±4.1)%,(26.1±3.6)%](P<0.05).The combined data suggest that the one-time centrifugation in vitro capacitation method and sperm floating capacitation method have better performance in improving IVF efficacy of bovine sexing semen,and the addition of 1.0%ascorbic acid in capacitation liquid and IVF liquid is recommended.In addition,the partial removal of cumulus cell from oocyte prior to IVF has important significance on improving the development of IVF embryo produced by sexing semen.