人诱导性多潜能干细胞(iPS细胞)系的建立  

Generation of A Human Induced Pluripotent Stem(iPS) Cells Line

作　　者：姚志芳[1] 史俊文[1] 贾俊双[1] 申红芬[1] 林晓琳[1] 肖高芳[1] 张晟[2] 肖东[1,2] 姚开泰[1] 

机构地区：[1]南方医科大学肿瘤研究所,中国广东广州510515 [2]南方医科大学比较医学研究所暨实验动物中心,中国广东广州510515

出　　处：《生命科学研究》2011年第6期512-518,共7页Life Science Research

基　　金：国家自然科学基金委员会-广东省联合基金重点项目(u0732006);广东省自然科学基金资助项目(9151063101000015);广东省科技计划项目(2009B060300008);南方医科大学优秀中青年科技人才库科研资助项目

摘　　要：掌握建立人iPS细胞系(induced pluripotent stem cells,iPSCs)的技术,以便为人肿瘤细胞重编程为iPS细胞建立技术平台.在人胚胎干细胞的培养条件下,通过携带Oct4、Sox2、c-Myc、Klf4 4个混合因子的慢病毒感染人皮肤成纤维细胞(CCD-1079SK细胞),从而诱导成干细胞样的克隆.根据人胚胎干细胞的特性进行如下鉴定:克隆形态、碱性磷酸酶活性、核型和CCD-1079SK细胞来源的克隆拟胚体(embryoid bodies,EBs)形成及分化等.结果显示,在人胚胎干细胞的培养环境中,导入Oct4、Sox2、c-Myc、Klf4 4个因子的CCD-1079SK细胞产生了一株iPSC克隆,这株iPSC克隆在细胞形态、增殖能力、胚胎细胞特异性表面抗原以及基因表达与人胚胎干细胞相似,此外,iPSC克隆在体外悬浮培养中形成拟胚体并分化成3个胚层.人iPS细胞系的成功建立为利用iPS细胞技术开展肿瘤细胞重编程研究奠定了坚实基础.To master the technology of reprogramming human somatic cells to iPS cells and set up a techni-cal platform to reprogram human cancer cells into iPS cells.Under human embryonic stem cell（hES cell） culture conditions,human skin fibroblasts（CCD-1079SK） cells infected by lentivirus mixture harboring Oct4,Sox2,c-Myc and Klf4 genes were induced into hES cell-like colonies,following characterization were done by comparatively analyzing hES cells markers： colony morphology,alkaline phosphatase（AP） activity,the expression profile of ES cell-marker genes,karyotype and embryoid body（EB）-mediated in vitro differentia-tion of CCD-1079SK cell-derived hES cell-like colonies.The results showed that human iPS cell line（iPSC-1） was generated from CCD-1079SK cells by introducing four genes,Oct4,Sox2,c-Myc and Klf4,under hES cell culture conditions.iPSC was similar to hES cells in morphology,proliferation,hES cell-specific surface anti-gens and gene expression.Additionally,iPSC could be cultured in suspension to form embryoid bodies（EBs） and differentiate into cell types of the three germ layers in vitro,in addition,human iPS cell line is successfully established,which will lay a solid foundation for performing cancer cell reprogramming by us-ing iPS cell technology.

关 键 词：人诱导性多潜能干细胞 肿瘤细胞重编程 肿瘤治疗 

分 类 号：R730.5[医药卫生—肿瘤]