DNAVEC Corporation, Tsukuba, Ibaraki 300-2611, Japan.
Proc Natl Acad Sci U S A. 2011 Aug 23;108(34):14234-9. doi: 10.1073/pnas.1103509108. Epub 2011 Aug 5.
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 nonpermissive 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.
在诱导多能干细胞（iPSC）的首次报道后，人们做出了相当大的努力来开发更有效的方法来生成无外源基因插入的 iPSC。在这里，我们表明，不具有整合到宿主基因组风险的 RNA 病毒载体仙台病毒载体是高效生成更安全 iPSC 的实用解决方案。我们通过引入温度敏感突变来改进仙台病毒载体，以便可以在非允许温度下轻松去除载体。使用这些载体，可以从人成纤维细胞和 CD34+脐带血细胞中高效产生无病毒/因子的 iPSC。温度转换处理在消除剩余的病毒载体相关基因方面更有效。所得的 iPSC 表达人类胚胎干细胞标志物，并表现出多能性。我们认为，从脐带血细胞中生成无转基因 iPSC 应该是将来提供同种异体 iPSC 衍生疗法的重要步骤。
