Laboratory of Gene Regulation, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
Laboratory of Gene Regulation, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
Stem Cell Reports. 2014 Nov 11;3(5):915-29. doi: 10.1016/j.stemcr.2014.08.014. Epub 2014 Oct 2.
The detailed mechanism of reprogramming somatic cells into induced pluripotent stem cells (iPSCs) remains largely unknown. Partially reprogrammed iPSCs are informative and useful for understanding the mechanism of reprogramming but remain technically difficult to generate in a predictable and reproducible manner. Using replication-defective and persistent Sendai virus (SeVdp) vectors, we analyzed the effect of decreasing the expression levels of OCT4, SOX2, KLF4, and c-MYC and found that low KLF4 expression reproducibly gives rise to a homogeneous population of partially reprogrammed iPSCs. Upregulation of KLF4 allows these cells to resume reprogramming, indicating that they are paused iPSCs that remain on the path toward pluripotency. Paused iPSCs with different KLF4 expression levels remain at distinct intermediate stages of reprogramming. This SeVdp-based stage-specific reprogramming system (3S reprogramming system) is applicable for both mouse and human somatic cells and will facilitate the mechanistic analysis of reprogramming.
重编程体细胞为诱导多能干细胞（iPSCs）的详细机制在很大程度上仍然未知。部分重编程的 iPSCs 对于理解重编程的机制具有信息性和有用性，但在可预测和可重复的方式下生成仍然具有技术难度。使用复制缺陷型和持续性的仙台病毒（SeVdp）载体，我们分析了降低 OCT4、SOX2、KLF4 和 c-MYC 表达水平的效果，发现低水平的 KLF4 表达可重复性地产生同质的部分重编程 iPSC 群体。KLF4 的上调允许这些细胞重新开始重编程，表明它们是暂停的 iPSC，仍然处于多能性的路径上。具有不同 KLF4 表达水平的暂停 iPSC 仍处于重编程的不同中间阶段。基于这种 SeVdp 的阶段特异性重编程系统（3S 重编程系统）适用于小鼠和人类体细胞，将有助于重编程的机制分析。
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