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Application of Oocytes In Vitro Maturation in Mice Biological Purification
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2019, 36(03):
53.
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Objective In mice biological purification,ovaries that did not ovulate after ovarian stimulation,were dissected and immature oocytes were isolated from follicles,then subjected to in vitro maturation for the ability of fertilization.By IVM,our aim was to improve oocytes utility ratios and as a remedial measure when conventional hyperstimulation fails. Method In mice biological purification,COCs were isolated by puncturing of ovarian follicles with a needle, that did not ovulate after PMSG and HCG stimulation, then subjected immediately to maturation in IVM culture.At the same time, the normal superovulation after PMSG and HCG injection, the immature oocytes in vitro maturation after PMSG injection, and suspected matured oocytes and nude oocytes in vitro maturation after PMSG and HCG injection were used as controls. After IVM and IVF, the 2-cell embryos were introduced to oviducts of the pseudo-pregnant foster mothers, and developed matured individuals in vivo. Result Ovaries that did not ovulate after PMSG and HCG injection(group A),the mature rate of immature oocytes was 87.0%±3.2%,2-cell rate was 55.1%±12.3%,blastocyst rate was 23.1%,41 2-cell embryos were introduced to oviducts of two pseudo-pregnant foster mothers, and 5 cubs were born, the birth rate was 12.2%。PMSG injection only, immature oocytes were obtained 48 h later(group B),in vitro maturation rate was 83.9%±3.9%, 2-cell rate was 51.8%±9.3%,blastocyst rate was 38.5%±13.9%.The normal superovulation after PMSG and HCG injection(group C),2-cell rate was 78.9%±0.6%,blastocyst rate was 78.0%±3.8%.The suspected matured oocytes and nude oocytes were obtained from ovaries that did not ovulate after PMSG and HCG injection(group D),matured in vitro 0 h,6h and 16～18 h,the mature rate,2-cell rate and blastocyst rate were all lower than the other three groups and had significant differences.The 2-cell rate of group A and group B had significant differences,and blastocyst rate had extremely significant differences compared with group C. Conclusion Oocytes IVM can be used as a remedial measure when conventional hyperstimulation fails, and also can be used to improve rare strains of mice oocytes utility ratios.
