Purpose: The biological purification of mice from non-ovulatory ovaries after hyperovulation involves collecting immature eggs from follicles, so that the egg cells can mature in vitro, have fertility and improve utilization. And as a traditional treatment for ovulation failure.
Method: In order to biologically purify mice, harvest the unovulated ovaries after injection of PMSG and HCG, cut the follicles under a solid microscope, and then select the mountain oocyte complex (COC) for maturation. Placed in droplets, and develop and mature in vitro. At the same time, the normal superovulation after PMSG and HCG injection, the in vitro maturation of immature eggs after PMSG injection, and the in vitro maturation of suspected mature eggs after PMSG and HCG injection were used as controls. After in vitro fertilization and in vitro embryo development, the two-cell embryo is transplanted into the recipient's oviduct and grows into a mature individual at the recipient.
Results: After PMSG and HCG were injected into the non-ovulatory group (group A), the in vitro maturation rate of oocytes was 87.0%±3.2%, the ratio of two cells was 55.1%±12.3%, and the ratio of scutellum follicles was 23.1%. 41 2-cell embryos were transferred. Two pups gave birth to 5 pups, and the number of litters was 12.2%. Only PMSG was injected, and the immature egg group was collected 48 hours later (group B). The in vitro maturation rate was 83.9%±3.9%, the two-cell rate was 51.8%±9.3%, and the scutellum rate was 38.5%±13.9%. In the normal high ovulation group (group C) injected with PMSG and HCG, the incidence of foreign diseases was 78.9%±0.6%, and the incidence of blastocyst cyst was 78.0%±3.8%. Inject PMSG and HCG into the non-ovulatory ovary, collect naked eggs and suspicious mature eggs (group D), in vitro maturation and culture for 0 hours, 6 hours, 16-18 hours, the maturation rate will be the ratio of the two cells in the other two groups, Compare the ratio of blastocysts. Low and very important difference. Compared with the normal control group C, there is a significant difference in the ratio of the two cells between the A group and the B group, as well as the ratio in the blastocyst sac.
Conclusion: In vitro maturation of oocytes (IVM) can be used to treat excessive ovulation failure in mouse biological purification, and can improve the utilization of rare mouse egg strains.