In present study the effects of EGF and GSH on the sheep in vitro fertilization of bovine were investigated, using M-199 as maturation control medium supplied with 5% OCS, using TALP medium as fertilization control medium supplied with 6%BSA, 5iu/ml heparine and 5mM caffine. The culture medium is SOFaa. When 10ng/mlEGF and 50ng/mlEGF is used in maturation medium, the the fertilization rate of EGF and control are 76.00%(114/150), 76.71% (112/146) and 60.70%(85/140),the blastcyst rate are 23.68%(27/114), 24.69%(20/81)and17.64%(15/85). The fertilization medium is TALP supplied with 6%BSA, 5iu/ml heparine and 5mM caffine. When 1mM GSH and 10mM is added, the fertilization rate of GSH and control is 71.66%(86/120), 63.28%(81/128) and 76.00%(114/150); the blastcyst rate is 34.88% (30/86), 24.69%(20/81)and 23.68%(27/114). The results indicate that M-199 supplement with 5% OCS and 10ng/mlEGF or 50ng/mlEGF can promote in vitro cleavage and development of bovine oocyte, but the higher concentration has no better effect than the lower concentration. At the same time, the fertilization medium added with 1mMGSH can promote in vitro development of bovine oocyte, without affect fertilization rate. But with the higher concentration of GSH, it cannot increase the rate of fertilization and blastcyst.