Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors
Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors
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DOI：
10.1073/pnas.1103509108
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发表时间：
2011-08-23
影响因子：
11.1
通讯作者：
Nishikawa, Shin-Ichi
中科院分区：
文献类型：
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作者：
Ban, Hiroshi;Nishishita, Naoki;Nishikawa, Shin-Ichi
自首次报道诱导多能干细胞（iPSCs）以来，人们一直在努力开发更有效的方法来产生不需要外源基因插入的iPSCs。本研究表明，仙台病毒载体是一种没有整合宿主基因组风险的RNA病毒载体，是一种有效生成更安全的iPSCs的实用解决方案。我们通过引入温度敏感突变改进了仙台病毒载体，使载体在非允许温度下可以很容易地去除。使用这些载体能够从人成纤维细胞和CD34（+）脐带血细胞中高效地生产无病毒/因子的iPSCs。温度转移处理在消除剩余的病毒载体相关基因方面更有效。由此产生的iPSCs表达人类胚胎干细胞标记物，并表现出多能性。我们认为，从脐带血细胞中产生无转基因的iPSCs应该是未来提供同种异体ipsc衍生治疗的重要一步。
After the first report of induced pluripotent stem cells (iPSCs), considerable efforts have been made to develop more efficient methods for generating iPSCs without foreign gene insertions. Here we show that Sendai virus vector, an RNA virus vector that carries no risk of integrating into the host genome, is a practical solution for the efficient generation of safer iPSCs. We improved the Sendai virus vectors by introducing temperature-sensitive mutations so that the vectors could be easily removed at non-permissive temperatures. Using these vectors enabled the efficient production of viral/factor-free iPSCs from both human fibroblasts and CD34(+) cord blood cells. Temperature-shift treatment was more effective in eliminating remaining viral vector-related genes. The resulting iPSCs expressed human embryonic stem cell markers and exhibited pluripotency. We suggest that generation of transgene-free iPSCs from cord blood cells should be an important step in providing allogeneic iPSC-derived therapy in the future.