EGFP小鼠ES细胞来源的生殖系嵌合小鼠的获得和鉴定

生殖系嵌合体的获得是实现ES细胞介导的转基因途径的决定步骤,而嵌合体的制作及生殖系嵌合体的获得则是判定ES细胞系是否具有配子分化能力的有效方法。利用一株表达绿色荧光蛋白的杂种ES细胞系制备出嵌合体小鼠,共获得9只表达绿色荧光蛋白的嵌合体小鼠,其中有8只雄性, 1只雌性,目前均发育成健康成年小鼠。流式细胞检测显示了绿色荧光蛋白在嵌合鼠以下器官的表达情况:心(77.96±15.78) %、脾(84.06±3.60) %、肾(42.49±19.79) %、骨髓(52.02±18.78) %。昆明雌鼠与雄性嵌合鼠杂交1代(F1)毛色表型分析显示该株ES细胞具有生殖系嵌合能力。

Germ-line Competent Chimeric Mice Derived from Es Cells of EGFP-transgenic Mouse

Generation of germline chimera is the key step in ES cell-mediated transgenesis, whereas the production of chimera, especially germline competent, is the only way to prove whether such potency is maintained. In this experiment, an established ES cell line expressing strong green fluorescence was used to produce chimera mice. After cell passage, ES cells were microinjected into KM's blastocysts. Nine EGFP-expressing chimeric mice were produced including eight male and one female. They developed into healthy adults. Results of a flow cytometry test showed the existence of GFP in several mice tissues: heart (77.96±15.78)%, spleen (84.06±3.60)%, kidney (42.49±19.79)% and marrow (52.02±18.78)%. The ES cell line proved to be important for the germ-line in contributing to the coat-color analysis of offspring(F1), produced by the cross between a female KM mouse and a male chimeric mouse.