2013年第40卷第5期目录
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封面故事:诱导多能干细胞(induced pluripotent stem cells,iPSCs)具有形成体内所有细胞类型的潜能,尤其是能以患者自身体细胞制备的iPSCs可用于特定疾病的治疗,不但能解决干细胞临床应用所遇到的免疫排斥问题和伦理医学等难题,而且在基础研究、再生医学以及药理科学中也有着重要的应用价值.为提高iPSCs的安全性,刘天庆领导的实验小组成功使用了携带Oct4、Sox2、Klf4和c-Myc基因的多顺反子质粒载体转染了脂肪干细胞,并将其重编程为iPSCs.经过细胞特异性标记基因和分化能力检测,证实iPSCs具有类似胚胎干细胞的形态与功能.DNA印迹实验显示质粒载体序列未整合至iPSCs基因组中,且保持正常核型.因此,本研究为产生具有高安全性的iPSCs提供了一个新方法.
(曲鑫建,刘天庆,宋克东,李香琴,葛 丹,关 水. 多顺反子质粒重编程人脂肪干细胞为诱导多能干细胞,本期第436~444页)
Cover Story: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 defined 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.
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综述与专论
研究快报
研究报告
技术与方法
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