Defining molecular cornerstones during fibroblast to iPS cell reprogramming in mouse
Defining molecular cornerstones during fibroblast to iPS cell reprogramming in mouse
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DOI：
10.1016/j.stem.2008.02.001
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发表时间：
2008-03-01
期刊：
影响因子：
23.9
通讯作者：
Hochedlinger, Konrad
中科院分区：
文献类型：
--
作者：
Stadtfeld, Matthias;Maherali, Nimet;Hochedlinger, Konrad
在成纤维细胞中异位表达转录因子Oct4、Sox2、c - Myc和Klf4可产生诱导多能干细胞（iPS细胞）。对于伴随核重编程的分子事件的性质和顺序知之甚少。利用强力霉素诱导型载体，我们已经表明外源性因子在大约10天内是必需的，之后细胞进入自我维持的多能状态。我们已经确定了在这个转变期之前和期间定义细胞群的标志物。虽然Thy1的下调以及随后SSEA - 1的上调发生在早期时间点，但内源性Oct4、Sox2、端粒酶的重新激活以及沉默的X染色体标志着重编程过程中的晚期事件。用这些标志物进行细胞分选能够显著富集有可能成为iPS细胞的细胞。我们的结果表明，因子诱导的重编程是一个渐进的过程，具有明确的中间细胞群，其中包含大多数准备成为iPS细胞的细胞。
Ectopic expression of the transcription factors Oct4, Sox2, c-Myc, and Klf4 in fibroblasts generates induced pluripotent stem (iPS) cells. Little is known about the nature and sequence of molecular events accompanying nuclear reprogramming. Using doxycycline-inducible vectors, we have shown that exogenous factors are required for about 10 days, after which cells enter a self-sustaining pluripotent state. We have identified markers that define cell populations prior to and during this transition period. While downregulation of Thy1 and subsequent upregulation of SSEA-1 occur at early time points, reactivation of endogenous Oct4, Sox2, telomerase, and the silent X chromosome mark late events in the reprogramming process. Cell sorting with these markers allows for a significant enrichment of cells with the potential to become iPS cells. Our results suggest that factor-induced reprogramming is a gradual process with defined intermediate cell populations that contain the majority of cells poised to become iPS cells.