中文摘要：

为建立多顺反子质粒载体转染技术获得人脂肪干细胞（adipose stem cells,ASCs）来源的诱导多能干细胞（inducedpluripotency stem cells,iPSCs）,应用2A元件连接Oct4/Sox2/KLF4/c-Myc四因子基因,构建为单一开放阅读框的多顺反子质粒载体.使用该质粒对ASCs进行转染及重编程为iPSC.采用形态学观察、特异性抗体免疫荧光鉴定、体外拟胚体诱导分化和体内畸胎瘤形成等方法进行鉴定.结果显示,ASCs成功重编程为iPSCs,具有与人胚胎干细胞相似的形态学及多向分化潜能;通过拟胚体和畸胎瘤实验证实iPSCs能在体内外分化成三胚层细胞;DNA印迹实验显示质粒载体序列未整合至iPSCs基因组中.因此,通过多顺反子质粒载体重编程技术成功建立的人iPSCs具有多向分化潜能,可减免发生插入突变和免疫排斥问题,为iPSCs在遗传性或退行性疾病的治疗奠定了实验基础.

英文摘要：

In order to establish a polycistronic plasmid delivery system that reprogram human adipose stem cells （ASCs） into induced pluripotency stem cells （iPSCs）, a polycistronic plasmid vector was constructed in which defned factors were fused in-frame into a single open reading frame via self-cleaving 2A sequences. The iPSCs were generated at 3 -4 weeks after ASCs have been transiently transfected with the polycistronic plasmid. Then, the iPSCs were identified subsequently via the morphological observation, immunofluorescence with specific antigen, embryoid body formation in vitro and teratoma formation in vivo. The results demonstrated that the characteristics of these generated iPSCs resembled embryonic stem cells （ESCs） in terms of the expression of pluripotent markers, and the ability to differentiate into the three embryonic germ layers in vitro by embryoid body generation together with in vivo by teratoma formation after injection into immunodeficient mice. Remarkably, Southern bolt revealed that the human iPSCs were not integrated with plasmid DNA sequence. Therefore, hASCs derived iPSCs has the pluripotency using polycistronic plasmid method, which provides a useful platform for the further study of hereditary or degenerative disease therapies with the potential to bypass both the insertional mutagenesis and immune rejection barriers.