Efficient iPS Cell Production with the MyoD Transactivation Domain in Serum-Free Culture
Efficient iPS Cell Production with the MyoD Transactivation Domain in Serum-Free Culture
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DOI：
10.1371/journal.pone.0034149
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发表时间：
2012-03-30
期刊：
影响因子：
3.7
通讯作者：
Kikyo, Nobuaki
中科院分区：
文献类型：
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作者：
Hirai, Hiroyuki;Katoku-Kikyo, Nobuko;Kikyo, Nobuaki
生产诱导多能干细胞(IPSCs)的一个主要困难是将分化的细胞重新编程为多能细胞的效率很低。我们先前的研究表明，当将Oct4和MyoD反式激活结构域(称为M30)与Sox2、Klf4和c-Myc(SKM)组成的融合基因与Sox2、Klf4和c-Myc(SKM)一起导入小鼠胚胎成纤维细胞(MEF)时，5%的MEF被重新编程为IPSCs。此外，M3O促进了大多数转导的MEF中多能基因的染色质重塑，包括未成为IPSCs的细胞。这些观察表明，在更有利的培养条件下，超过5%的细胞可能已经获得了成为IPSCs的能力。在这里，我们通过在无血清培养液中低密度培养转导细胞，将M30-SKM用于制备小鼠IPSCs的效率提高到26%。相反，在相同的培养条件下，野生型Oct4和SKM(OSKM)的组合效率从0.1%提高到2%。对于人IPSCs，在类似的无血清培养条件下，M30-SKM的效率为7%，而OSKM的效率为1%。这项研究强调了将MyoD的反式激活结构域与良好的培养环境相结合的力量。
A major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gene composed of Oct4 and the transactivation domain of MyoD (called M3O), along with Sox2, Klf4 and c-Myc (SKM). In addition, M3O facilitated chromatin remodeling of pluripotency genes in the majority of transduced MEFs, including cells that did not become iPSCs. These observations suggested the possibility that more than 5% of cells had acquired the ability to become iPSCs given more favorable culture conditions. Here, we raised the efficiency of making mouse iPSCs with M3O-SKM to 26% by culturing transduced cells at low density in serum-free culture medium. In contrast, the efficiency increased from 0.1% to only 2% with the combination of wild-type Oct4 and SKM (OSKM) under the same culture condition. For human iPSCs, M3O-SKM achieved 7% efficiency under a similar serum-free culture condition, in comparison to 1% efficiency with OSKM. This study highlights the power of combining the transactivation domain of MyoD with a favorable culture environment.