摘要
目的对DPF2干扰慢病毒感染H9细胞,获得稳定的人胚胎干细胞克隆细胞系的方法学探讨。方法将各型带有GFP荧光标签的DPF2-RNAi慢病毒导入H9细胞中,嘌呤霉素筛选前后,相差和荧光显微镜下观察细胞形态和细胞感染效率;Western blot鉴定感染后的各组H9细胞系内DPF2蛋白表达水平。结果成功获得慢病毒感染后的H9稳定细胞系,成功筛选出#25540 DPF2-RNAi慢病毒,其沉默或敲低H9细胞的效率更高,稳定性更好。结论应用携带DPF2-RNAi的慢病毒感染人胚胎干细胞,使目标基因沉默,探索建立稳定细胞系的方法,为进一步研究提供细胞模型。
Objective To investigate the method of obtaining stable clonal cell lines of human embryonic stem cells by infecting H9 cells with DPF2 interfered lentivirus.Methods Various types of DPF2-RNAi lentivirus with GFP fluorescent tags were introduced into H9 cells.Before and after puromycin screening,the cell morphology and cell infection efficiency were observed under phase contrast and fluorescence microscope.Western blot was used to identify DPF2 protein expression level in each group of H9 cell lines after infection.Results The stable H9 cell line after lentivirus infection was successfully obtained.Among the three groups,#25540 DPF2-RNAi was successfully obtained,the lentivirus silencing or knocking down H9 cells is more efficient and stable.Conclusions The lentivirus carrying DPF2-RNAi was used to infect human embryonic stem cells to silence the target gene,explore the method of establishing a stable cell line,and provide a cell model for further research.
作者
张涤娟
刘超
ZHANG Di-juan(Reproductive medicine department,the first affiliated hospital&Yijishan hospital of Wannan Medical College,Wuhu,Anhui,241002,China)
出处
《齐齐哈尔医学院学报》
2021年第11期932-935,共4页
Journal of Qiqihar Medical University
基金
国家自然科学基金面上项目(31271159)
皖南医学院中青年科研基金项目(WK2020F31)。
