红色荧光蛋白标记的小鼠胚胎干细胞系建立及体内成瘤的初步研究  

Establishment of Mouse Embryonic Stem Cell Line Marked by Red Fluorescent Protein and Primary Study on Its Tumor in Vivo

作　　者：赵无恙[1] 刘韬[2] 钱其军[1,2] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018 [2]第二军医大学东方肝胆外科医院病毒治疗实验室,上海200438

出　　处：《浙江理工大学学报（自然科学版）》2015年第2期244-248,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家杰出青年科学基金项目(30925037)

摘　　要：建立能够稳定表达红色荧光的小鼠胚胎干细胞系,并研究其在体内不同部位形成畸胎瘤的能力。构建含红色荧光蛋白(DsRed-Express)的表达质粒,并采用磷酸钙法包装成慢病毒转染小鼠胚胎干细胞(mESCs)。筛选得到强阳性克隆,对其进行RT-PCR检测和裸鼠、C57小鼠体内成瘤实验验证细胞特性,最后获得稳定表达红色荧光的胚胎干细胞(Red-ESCs)。Red-ESCs能够表达多能性基因,如Oct4,Sox2,Nanog等,也能在裸鼠体内成瘤。不同部位的畸胎瘤在基因表达上有差异,其中肝部形成的畸胎瘤表达肝特性基因。成功建立稳定表达红色荧光的小鼠胚胎干细胞系,其在体内形成的畸胎瘤性质受到体内环境的影响。This paper establishes a mouse embryonic stem cell line which can express red fluorescent stably and studies its ability to form teratoma in different parts in vivo.DsRed-Express plasmid is constructed.Calcium phosphate method is used to pack it into lentiviral transfection mouse embryonic stem cells(mESCs).Strong positive clone is gained through screening.The cell characteristics are tested by RT-PCR test and tumor formation experiment in nude mice and C57 mice.Finally,Red-ESCs which can express red fluorescent stably are gained.Red-ESCs can express pluripotent genes,such as Oct4,Sox2 and Nanog,and form tumor in nude mice.Teratoma in different parts has differences in gene expression.Teratoma formed in liver expresses hepatic specific gene.Mouse embryonic stem cell line which can express red fluorescent stably is established successfully.Teratoma nature is influenced by internal environment.

关 键 词：胚胎干细胞 红色荧光蛋白 畸胎瘤 肝特异性基因 

分 类 号：Q256[生物学—细胞生物学]