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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Synergic reprogramming of mammalian cells by combined exposure to mitotic Xenopus egg extracts and transcription factors
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Synergic reprogramming of mammalian cells by combined exposure to mitotic Xenopus egg extracts and transcription factors

机译：通过联合接触有丝分裂爪蟾卵提取物和转录因子对哺乳动物细胞进行协同重编程

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摘要

Transfer of somatic cell nuclei to enudeated eggs and ectopic expression of specific transcription factors are two different reprogramming strategies used to generate pluripotent cells from differentiated cells. However, these methods are poorly efficient, and other unknown factors might be required to increase their success rate. Here we show that Xenopus egg extracts at the metaphase stage (M phase) have a strong reprogramming activity on mouse embryonic f ibroblasts (MEFs). First, they reset replication properties of MEF nuclei toward a replication profile characteristic of early development and they erase several epigenetic marks, such as trimethylation of H3K9, H3K4, and H4K20. Second, when MEFs are reversibly permea-bilized in the presence of M-phase Xenopus egg extracts, they show a transient increase in cell proliferation, form colonies, and start to express specific pluripotency markers. Finally, transient exposure of MEF nuclei to M-phase Xenopus egg extracts increases the success of nuclear transfer to enucleated mouse oocytes and strongly syner-gizes with the production of pluripotent stem cells by ectopic expression of transcription factors. The mitotic stage of the egg extract is crucial, because none of these effects is detected when using inter-phasic Xenopus egg extracts. Our data demonstrate that mitosis is essential to make mammalian somatic nuclei prone to reprogramming and that, surprisingly, the heterologous Xenopus system has features that are conserved enough to remodel mammalian nuclei.
机译：体细胞核转移至枚举的卵和特异转录因子的异位表达是用于从分化细胞中产生多能细胞的两种不同的重编程策略。但是，这些方法效率低下，可能需要其他未知因素来提高其成功率。在这里，我们显示爪蟾卵提取物在中期（M期）对小鼠胚胎成纤维细胞（MEF）具有很强的重编程活性。首先，他们将MEF核的复制特性重设为早期发育的复制特征，并且擦除了一些表观遗传标记，例如H3K9，H3K4和H4K20的三甲基化。其次，当MEF在M期爪蟾卵提取物的存在下可逆渗透时，它们显示出细胞增殖的短暂增加，形成菌落，并开始表达特定的多能性标记。最后，MEF核对M期非洲爪蟾卵提取物的短暂暴露增加了核转移至去核小鼠卵母细胞的成功率，并通过异位表达转录因子与多能干细胞的产生产生了强大的协同作用。卵提取物的有丝分裂阶段至关重要，因为在使用相间非洲爪蟾卵提取物时，没有检测到这些作用。我们的数据表明，有丝分裂对于使哺乳动物的体细胞核易于重编程至关重要，而且，令人惊讶的是，异源非洲爪蟾系统具有足够保守的特征，可以重塑哺乳动物的细胞核。

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    Olivier Gamer; Stephane Bocquet; Isabelle Peiffer; Vincent Brochard; Philippe Arnaud; Aurore Puy; Alice Jouneau; Robert Feil; Jean-Paul Renard; Marcel Mechali;

  • 作者单位

    Centre National de la Recherche Scientifique (CNRS), Institute of Human Genetics, 34396 Montpellier, France;

    Centre National de la Recherche Scientifique (CNRS), Institute of Human Genetics, 34396 Montpellier, France;

    Centre National de la Recherche Scientifique (CNRS), Institute of Human Genetics, 34396 Montpellier, France;

    lnstitut National de la Recherche Agronomique (INRA), Unite Mixte de Recherche 1198 Biologie du Developpement et Reproduction, F-78352 Jouy-en-Josas, France;

    lnstitut de Genetique Moleculaire de Montpellier, Unite Mixte de Recherche 5535, 34000 Montpellier, France;

    Centre National de la Recherche Scientifique (CNRS), Institute of Human Genetics, 34396 Montpellier, France;

    lnstitut National de la Recherche Agronomique (INRA), Unite Mixte de Recherche 1198 Biologie du Developpement et Reproduction, F-78352 Jouy-en-Josas, France;

    lnstitut de Genetique Moleculaire de Montpellier, Unite Mixte de Recherche 5535, 34000 Montpellier, France;

    lnstitut National de la Recherche Agronomique (INRA), Unite Mixte de Recherche 1198 Biologie du Developpement et Reproduction, F-78352 Jouy-en-Josas, France;

    Centre National de la Recherche Scientifique (CNRS), Institute of Human Genetics, 34396 Montpellier, France;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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