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General Research Article

两种转录因子组合对人表皮黑素细胞诱导的多能干细胞的表征。

卷 19, 期 6, 2019

页: [395 - 403] 页: 9

弟呕挨: 10.2174/1566523220666200211105228

价格: $65

摘要

目的：为了更有效地产生诱导性多能干细胞（iPSC），至关重要的是要确定易于进入且可能需要较少因子转化为iPSC的体细胞。 方法：用含有3种转录因子（OCT-4，KLF-4和c-MYC，3F）或4种转录因子（OCT-4，KLF-4，c-MYC和SOX-2）的慢病毒载体转导人表皮黑素细胞， 4F）。一旦克隆形成，就进行与干细胞多能性有关的测定，包括碱性磷酸酶染色，DNA甲基化水平，干细胞标志物的表达和超微结构分析。然后诱导获得的iPSCs在体外分化为代表三个胚胎层的细胞。 结果：在用3F或4F转导表皮黑素细胞后的7天，形成了碱性磷酸酶染色呈阳性的克隆。用针对OCT-4和SOX-2的抗体进行的荧光染色呈强阳性，并且细胞显示出高的细胞核质比和活跃的核运动。通过超微结构分析在细胞质中未发现黑素体。克隆细胞与其亲代细胞之间的DNA甲基化水平存在明显差异。但是，在3F或4F转染的克隆细胞之间没有显着差异。同时，iPSCs在体外成功分化为三个胚层细胞。 结论：人类表皮黑素细胞不需要异位SOX-2表达即可转化为iPSC，并且可以作为获得具有较少遗传成分的患者特异性iPSC的替代来源。

关键词: 表皮，黑素细胞，多能，重编程，iPSCS，转录。

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图形摘要

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