Objective AG129 mice are difficult to give birth naturally, and conventional in vitro fertilization reagents cannot be used to obtain enough embryos for transplantation. Reduced glutathione (GSH) was used to adjust the composition of in vitro fertilization fluid of mice, and the in vitro fertilization of AG129 mice was completed. It provides ideas for biological purification and rapid propagation of genetically engineered mice of difficult in vitro fertilization strains.
Methods Three experimental groups containing sperm capacitation and fertilization fluid with different concentrations of GSH were designed, with concentrations of 0.5, 1, and 2 mmol/L, respectively, and a control group containing no GSH sperm capacitation fluid and fertilization fluid. 5 mice were superovulated and repeated 3 times.
Results There were significant differences in the average fertilization rate between different GSH experimental groups and control groups (P < 0.05). When the GSH concentration in sperm capacitation and fertilization fluid was 1 mmol/L, the in vitro fertilization effect of AG129 mice was the best. The fertilization rate was (43.35±1.76)%, and the embryos were in good condition. The recipient mice for embryo transfer were obtained. All recipient animals were pregnant and gave birth smoothly.
Conclusion When the concentration of GSH in in vitro fertilization fluid is 1 mmol/L, it can significantly improve the efficiency of in vitro fertilization in AG129 mice, and provide a solution for the biological purification and rapid reproduction of mice similar to in vitro fertilization difficult strains.