Nakagawa Masato, Koyanagi Michiyo, Tanabe Koji, Takahashi Kazutoshi, Ichisaka Tomoko, Aoi Takashi, Okita Keisuke, Mochiduki Yuji, Takizawa Nanako, Yamanaka Shinya
Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
Nat Biotechnol. 2008 Jan;26(1):101-6. doi: 10.1038/nbt1374. Epub 2007 Nov 30.
Direct reprogramming of somatic cells provides an opportunity to generate patient- or disease-specific pluripotent stem cells. Such induced pluripotent stem (iPS) cells were generated from mouse fibroblasts by retroviral transduction of four transcription factors: Oct3/4, Sox2, Klf4 and c-Myc. Mouse iPS cells are indistinguishable from embryonic stem (ES) cells in many respects and produce germline-competent chimeras. Reactivation of the c-Myc retrovirus, however, increases tumorigenicity in the chimeras and progeny mice, hindering clinical applications. Here we describe a modified protocol for the generation of iPS cells that does not require the Myc retrovirus. With this protocol, we obtained significantly fewer non-iPS background cells, and the iPS cells generated were consistently of high quality. Mice derived from Myc(-) iPS cells did not develop tumors during the study period. The protocol also enabled efficient isolation of iPS cells without drug selection. Furthermore, we generated human iPS cells from adult dermal fibroblasts without MYC.
体细胞的直接重编程为生成患者特异性或疾病特异性多能干细胞提供了契机。通过逆转录病毒转导四种转录因子：Oct3/4、Sox2、Klf4和c-Myc，从小鼠成纤维细胞中生成了此类诱导多能干细胞（iPS细胞）。小鼠iPS细胞在许多方面与胚胎干细胞（ES细胞）难以区分，并能产生具有生殖系能力的嵌合体。然而，c-Myc逆转录病毒的重新激活会增加嵌合体和后代小鼠的致瘤性，从而阻碍临床应用。在此，我们描述了一种生成iPS细胞的改良方案，该方案无需使用Myc逆转录病毒。采用此方案，我们获得的非iPS背景细胞显著减少，且所生成的iPS细胞始终具有高质量。在研究期间，源自Myc(-) iPS细胞的小鼠未发生肿瘤。该方案还能够在无药物筛选的情况下高效分离iPS细胞。此外，我们从成人皮肤成纤维细胞中生成了不含MYC的人iPS细胞。
