小鼠胚胎成纤维细胞的转染与阳性细胞的初步筛选  

Transfer of mouse fetal fibroblasts and primary selection of positive cells

作　　者：姜秋[1] 倪雪岩[2] 聂代邦[3] 孙宏晨[1] 

机构地区：[1]吉林大学口腔医院儿童口腔科,吉林长春130041 [2]首都医科大学附属北京安贞医院口腔医疗中心,北京100029 [3]吉林大学畜牧兽医学院生物技术系,吉林长春130062

出　　处：《吉林大学学报（医学版）》2008年第6期990-993,共4页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(200705347)

摘　　要：目的:建立表达白血病抑制因子(LIF)的胚胎成纤维细胞的方法体系,为建立转基因动物模型提供理论依据。方法:以13.5dICR小鼠胚胎为材料,采用胰酶消化方法,培养小鼠原代胚胎成纤维细胞。利用脂质体介导方法,将线性化的pcDNA3.1-LIF真核表达载体转染至小鼠胚胎成纤维细胞中。经G418筛选阳性细胞,提取阳性细胞的基因组DNA并测序。结果:通过对小鼠原代胚胎成纤维细胞的分离、培养,获得原代胚胎成纤维细胞。经过转染,建立了表达LIF的胚胎成纤维细胞,并在倒置显微镜下观察到筛选得到的阳性细胞。对阳性细胞克隆质粒测序表明同源性达99%。结论:pcDNA3.1-LIF真核表达载体成功转染到小鼠胚胎成纤维细胞中。Objective To establish the method to express leukemia inhibitory factor(LIF) in fetal fibroblasts in order to provide theoretical foundation for establishment of transgenic animal model.Methods Using the fetal of 13.5 d ICR mouse,the primary fetal fibroblasts were cultivated by trypsin enzyme digestion.The lineared eukaryotic expression vector pcDNA3.1-LIF was transferred to the fetal fibroblasts of mouse with liposome induction.The positive cells were selected by G418,genome DNA of the positive cells was e...

关 键 词：白血病抑制因子 成纤维细胞 转染 小鼠 

分 类 号：R363[医药卫生—病理学]