Activation Of KB Mouse Oocytes And Their Parthenogenetic Development In Vitro

KB mouse, which is bred in our country by near inbred, has been a good trial animal model because of the well-known inheriting background, short reproduction cyc1e and the strong environmental adapt ability. In this experiment, we isolated endometrial epithelium cells from KB mouse, and studied the activation of mouse oocytes and culturing of parthenogenesis. We also discussed some factors of activation and development. Our work may be useful to further isolate and esTab. lish mouse parthenogenesis ES cell lines. The main contents are as follows:1. Endometrial epithelium cells were isolated by the use of 2.5 mg/mLTrypsine + 0.04% EDTA in 37℃to digest the endometrial epithelium in 30 min, combined with scraping method. The vitality and amount of cells was the highest and the cells are more single-typed, the vitality of epithelium cells declined with the increase of passages (detected by MTT method).2. Mouse oocytes were activated by 7% ethanol. After 5-7 min ,the activated effect and the parthenogenetic development were better than others, the development rates of over morula and blastocyst were up to 29.77%.3. When mouse oocytes were activated by SrCl2, the optimum concentration was 6-10 mmol/L and the time of the optimum activated is 3-6 h, so the development rates of over morula and blastocyst was up to 16.67%.4. In combined activation, mouse oocytes were activated 5 min by 7% ethanol, and was cultured 3 h into medium which combined with 2 mmol/L 6-DMAP +5μg/mLCB, the development rates of over morula and blastocyst were up to 35.77%.5. CZB and mM16 medium containing 15%FBS supported optimum development from 2- cell embryos to blastocyst(27.2%,26.3%). In the meantime, compared with the development rates of blastocyst in CZB and mM16 medium containing 15%FBS with containing BSA,the difference were extreme.6. For mouse parthenogenesis embryos, in CZB medium, 41.66% embryos prevented the 2- cell block and finally 29.17% embryos reached morula or blastocyst.But7.45% embryos developed to 3～4-cell embryos in mM16 medium, exhibiting an obvious"2-cell block".7. Activared MⅡoocytes prevented the 2-cell block in CZB+15%FBS medium and developed to morula or blastocyst. The presence of 1.1mg/mL glucose in the medium after culturing 48 h improved the continuous development of embryos.