Objective In the biological purification of mice, from the unovulated ovaries after superovulation, the immature eggs in the follicles are obtained by in vitro maturation of oocytes to make them mature in vitro and have the ability to fertilize, so as to improve the utilization of eggs and promote superovulation as a routine method. A remedy for failure.
Methods In the biological purification of mice, the non-ovulatory ovaries after injection of PMSG and HCG were taken, and the follicles were cut under a solid microscope to select cumulus oocyte complexes (COCs), which were placed in maturation droplets to mature in vitro. At the same time, normal superovulation after injection of PMSG and HCG, immature eggs obtained after injection of PMSG only, and suspected mature eggs and manipulated eggs after injection of PMSG and HCG were used as controls. After in vitro fertilization and in vitro embryo development, the 2-cell embryos are transplanted into the recipient's fallopian tube, allowing them to develop into mature individuals in the recipient.
Results In the non-ovulatory group (group A) injected with PMSG and HCG, the in vitro maturation rate of oocytes was 87.0%±3.2%, the two-cell rate was 55.1%±12.3%, and the blastocyst rate was 23.1%. 41 two-cell embryos were transferred to 2 Only recipient mice, 5 pups were born, and the litter rate was 12.2%. Only PMSG was injected, and the immature egg group (group B) was taken after 48 hours. The in vitro maturation rate was 83.9%±3.9%, the two-cell rate was 51.8%±9.3%, and the blastocyst rate was 38.5%±13.9%. In the normal superovulation group (group C) injected with PMSG and HCG, the two-cell rate was 78.9%±0.6%, and the blastocyst rate was 78.0%±3.8%. Injected PMSG and HCG to anovulatory ovaries, took naked eggs and suspected mature eggs (group D), and cultured them in vitro for 0h, 6h and 16-18h. The maturation rate, double cell rate and blastocyst rate were compared with the other three groups. low and very significant difference. Compared with the normal control group C, the two-cell rate and the blastocyst rate were significantly different in groups A and B.
Conclusions In vitro oocyte maturation (IVM) can be used as a remedy for the failure of superovulation-stimulating in mouse biopurification, and can improve egg utilization in rare strains of mice.