Effect Of Different Additives On The IVM/IVF Of Bovine Oocytes And Culture Of Zygotes

Using ovaries of bovine collected from slaughter houses, IVM (in vitro maturation) and IVF (in vitro fertilization) of 2-6mm oocytes and culture of early embryos were studied, and the results are as follows:1. TCM-199 is the best basic culture medium for the maturation rate of oocytes cultured in this medium (81.25% ) being significantly higher than those in DMEM (61.13%) or in M16 (66.78%) .respectively.2. The maturation rates of oocytes cultured in TCM-199 with NCS ( new calf serum ), human serum and BSA (bovine serum albumin) (73.38, 86.50 and 46.62%, respectively) are significantly different, showing that human serum is more favorable to the IVM of bovine oocytes and will be a prospective additive of culture medium for oocyte maturation.3. 64.86, 96.55, and 84.73% of the maturation rates for oocytes cultured in TCM-199 medium with 5, 10 and 15% human serum respectively (significantly different between each other) suggest that 10% human serum is more suitable to IVM of oocytes, and 57.65 53.42, 42.50% of the cleavage rates of oocytes cultured in TCM-199 with 10, 15, 5% human serum, respectively (significant difference between 10 and 5% or between 15 and 5% human serum, while no difference between the former two human serums) indicate that 10, 15% human serum may be used for IVF and IVC.4. No significant difference exists in the maturation rates of bovine oocytes cultured in medium with or without insulin, indicating insulin is not necessary for oocyte culture.5. There is no significant difference between the cleavage rates of embryos fertilized with the sperm capacitated with mTyrode's and BO solution respectively, suggesting that both of them could be used for the sperm capacitation.6. After sperm capacitating in the capacitation solution with Caffein, Heparin and Caffein+ Heparin respectively, the cleavage rates of oocytes are 53.42, 49.02 and 56.15%, respectively, with no significant difference exiting between the first and the latter twosolutions, but a significant difference between the latter two capacitation solution, revealing that Caffein and Heparin can cooperate with each other and improve the motility of sperm.7. In the development of early embryo, the rates of morula and blatocyst are 5.60,10.77 and 21.43%, respectively, cultured in the three mediums with or without glucose (medium 1: with glucose; 2: without glucose; 3: without glucose in the preliminary culture stage followed by culture with glucose), giving the result that glucose is an essential energy factor in late development of embryo, although it represses the development of early embryo.