Utikal Jochen, Maherali Nimet, Kulalert Warakorn, Hochedlinger Konrad
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Massachusetts General Hospital Center for Regenerative Medicine, 185 Cambridge Street, Boston, MA 02114, USA.
J Cell Sci. 2009 Oct 1;122(Pt 19):3502-10. doi: 10.1242/jcs.054783. Epub 2009 Sep 1.
Induced pluripotent stem cells (iPSCs) have been derived at low frequencies from different cell types through ectopic expression of the transcription factors Oct4 and Sox2, combined with either Klf4 and c-Myc or Lin28 and Nanog. In order to generate iPSCs more effectively, it will be crucial to identify somatic cells that are easily accessible and possibly require fewer factors for conversion into iPSCs. Here, we show that both human and mouse melanocytes give rise to iPSCs at higher efficiencies than fibroblasts. Moreover, we demonstrate that a mouse malignant melanoma cell line, which has previously been reprogrammed into embryonic stem cells by nuclear transfer, remains equally amenable to reprogramming into iPSCs by these transcription factors. In contrast to skin fibroblasts, melanocytes and melanoma cells did not require ectopic Sox2 expression for conversion into iPSCs. iPSC lines from melanocytic cells expressed pluripotency markers, formed teratomas and contributed to viable chimeric mice with germ line transmission. Our results identify skin melanocytes as an alternative source for deriving patient-specific iPSCs at increased efficiency and with fewer genetic elements. In addition, our results suggest that cancer cells remain susceptible to transcription factor-mediated reprogramming, which should facilitate the study of epigenetic changes in human cancer.
通过转录因子Oct4和Sox2的异位表达,联合Klf4和c-Myc或Lin28和Nanog,已从不同细胞类型中以低频率诱导出多能干细胞(iPSC)。为了更有效地生成iPSC,识别易于获取且可能转化为iPSC所需因子较少的体细胞至关重要。在此,我们表明,人类和小鼠黑素细胞产生iPSC的效率均高于成纤维细胞。此外,我们证明,一种先前已通过核移植重编程为胚胎干细胞的小鼠恶性黑色素瘤细胞系,同样易于被这些转录因子重编程为iPSC。与皮肤成纤维细胞不同,黑素细胞和黑色素瘤细胞转化为iPSC不需要异位表达Sox2。来自黑素细胞的iPSC系表达多能性标志物,形成畸胎瘤,并产生具有种系传递能力的活的嵌合小鼠。我们的结果确定皮肤黑素细胞是一种以更高效率和更少遗传元件获得患者特异性iPSC的替代来源。此外,我们的结果表明癌细胞仍然易受转录因子介导的重编程影响,这将有助于研究人类癌症中的表观遗传变化。