Study On Parthenogenetic Activation And Enucleation Of KM Mouse

Since 1998, after the first successful mouse somatic cloning, the mouse somatic cell nucleus transplant technology was perfect gradually, many strain's mice have all been successfu in the somatic cell nucleus transplant, obtained the clone descendant, but KM mouse has not been the successful report actually. The KM mouse took our country one kind of important mouse strain, has many merits, such as nicer heredity background, short reproductive cycle, strong environment adaptablity and so on..It is widely applies in the pharmacology, the toxicology, the microbiology research as well as the drugs, the biological preparations examination, is a excellence experimental animal model.The study has conducted the research from 3 aspects to KM mouse somatic cell nucleus transplant: Selection of good cultivation medium for zygote in vitro; activates the method differently and goes to the nuclear method to the mouse ovocyte orphaned female growth influence the research. effect of differerent activation methods on embryo ectogenesis; method of oocyte enucleated on cloning.1. In vitro, cultivation environment has remarkable influence on mouse embryo growth before planting, many strain mouse embryo displays growth standstill on 2- cell obviously, including KM mouse. Therefore, finding a cultivation system which overcomes growth standstill would be extremely necessary. This experiment based on the fact which zygotes altogether cultivated with mouse oviduct epithelial cell, compared with zygote growth ability in mouse male pronucleus formative in cultivation medium CZB, M16, KSOM, mCZB, mM16, in mKSOM. And obtains a kind of cultivation medium effectively to support the KM mouse zygote in vitro growth, lays the foundation for the restructuring embryo culture. The experimental result indicated that, the cultivation system can obviously promot growth of early time embryo. Improvement M16, the KSOM cultivation meduim can satisfy the request of early time embryo growth, and obtain the high blastocyst rate, that respectively is 80.23%, 78.82%. they have qualification as nultivation meduim for restructuring embryo.2. Activation project is selected from 4 kinds of activate method:oocytes were activated 6～9min by 7% ethanol; activation meduim M16 including 2, 5, 8, 10mmol/ML SrCl₂ plus 5μg/mL CB activated oocytes in five hours; activation meduim M16 including 10mmol/mL SrCl₂ plus 5μg/ML CB activated oocytes...