Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts
Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts
复制标题
DOI：
10.1038/nbt1374
复制
发表时间：
2008-01-01
影响因子：
46.9
通讯作者：
Yamanaka, Shinya
中科院分区：
文献类型：
--
作者：
Nakagawa, Masato;Koyanagi, Michiyo;Yamanaka, Shinya
体细胞的直接重编程提供了产生患者或疾病特异性多能干细胞的机会。通过逆转录病毒转导四种转录因子：Oct 3/4、Sox 2、Klf 4和c-Myc，从小鼠成纤维细胞中产生这种诱导多能干细胞（iPS）（1）。小鼠iPS细胞在许多方面与胚胎干（ES）细胞不可区分，并产生生殖系感受态嵌合体（2-4）。然而，c-Myc逆转录病毒的再活化增加了嵌合体和后代小鼠的致瘤性，阻碍了临床应用（3）。在这里，我们描述了一个修改后的协议，不需要Myc逆转录病毒的iPS细胞的生成。通过该方案，我们获得了显著更少的非iPS背景细胞，并且产生的iPS细胞始终具有高质量。来自Myc-iPS细胞的小鼠在研究期间没有发生肿瘤。该方案还能够在没有药物选择的情况下有效分离iPS细胞。此外，我们从不含MYC的成人真皮成纤维细胞产生人iPS细胞。
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(1). Mouse iPS cells are indistinguishable from embryonic stem (ES) cells in many respects and produce germline- competent chimeras(2-4). Reactivation of the c-Myc retrovirus, however, increases tumorigenicity in the chimeras and progeny mice, hindering clinical applications(3). 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.