DNA synthesis is required for reprogramming mediated by stem cell fusion.
DNA synthesis is required for reprogramming mediated by stem cell fusion.
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DOI：
10.1016/j.cell.2013.01.012
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发表时间：
2013-02-14
期刊：
影响因子：
64.5
通讯作者：
Fisher AG
中科院分区：
文献类型：
--
作者：
Tsubouchi T;Soza-Ried J;Brown K;Piccolo FM;Cantone I;Landeira D;Bagci H;Hochegger H;Merkenschlager M;Fisher AG
胚胎干细胞（ESC）可以通过一种快速但知之甚少的机制指导分化细胞在细胞间融合后向多能性转化。在这里，我们使用离心淘洗来富集细胞周期连续阶段的小鼠 ESC，并表明 S/G2 期的 ESC 在异核和混合测定中显性重编程淋巴细胞和成纤维细胞的能力增强。重编程的成功与诱导异核体中体细胞伙伴核内核苷酸早熟的能力相关。 BrdU 脉冲标记实验表明，几乎所有基于 Oct4 重新表达而成功重编程的体细胞核，都在与 ESC 融合后的 24 小时内进行了 DNA 合成。这对于成功重编程至关重要，因为抑制 DNA 聚合酶活性的药物可以有效阻止多能转化。这些数据表明，核苷酸掺入是实验性 ESC 异核体体细胞表观遗传重编程的早期且关键的事件。 ► 逆流离心淘洗可富集 ESC 中的特定细胞周期阶段 ► 富集 S/G2 的 ESC 具有增强的重编程体细胞的能力 ► DNA 合成对于 ESC 融合介导的体细胞重编程至关重要 小鼠胚胎干细胞 (ESC) 重编程异核体内体细胞的能力需要诱导体细胞核中的 DNA 合成，这表明 DNA 复制对于体细胞成功转化为体细胞至关重要。多能性。
Embryonic stem cells (ESCs) can instruct the conversion of differentiated cells toward pluripotency following cell-to-cell fusion by a mechanism that is rapid but poorly understood. Here, we used centrifugal elutriation to enrich for mouse ESCs at sequential stages of the cell cycle and showed that ESCs in S/G2 phases have an enhanced capacity to dominantly reprogram lymphocytes and fibroblasts in heterokaryon and hybrid assays. Reprogramming success was associated with an ability to induce precocious nucleotide incorporation within the somatic partner nuclei in heterokaryons. BrdU pulse-labeling experiments revealed that virtually all successfully reprogrammed somatic nuclei, identified on the basis of Oct4 re-expression, had undergone DNA synthesis within 24 hr of fusion with ESCs. This was essential for successful reprogramming because drugs that inhibited DNA polymerase activity effectively blocked pluripotent conversion. These data indicate that nucleotide incorporation is an early and critical event in the epigenetic reprogramming of somatic cells in experimental ESC-heterokaryons. ► Counterflow centrifugal elutriation enriches for specific cell-cycle stages in ESCs ► S/G2-enriched ESCs have an enhanced capacity to reprogram somatic cells ► DNA synthesis is critical in fusion-mediated reprogramming of somatic cells by ESCs The capacity of mouse embryonic stem cells (ESCs) to reprogram somatic cells within heterokaryons requires induction of DNA synthesis in the somatic nucleus, suggesting that DNA replication is essential for the successful conversion of somatic cells toward pluripotency.
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影响因子：
46.9
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Deng, Jie;Shoemaker, Robert;Xie, Bin;Gore, Athurva;LeProust, Emily M.;Antosiewicz-Bourget, Jessica;Egli, Dieter;Maherali, Nimet;Park, In-Hyun;Yu, Junying;Daley, George Q.;Eggan, Kevin;Hochedlinger, Konrad;Thomson, James;Wang, Wei;Gao, Yuan;Zhang, Kun
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通讯作者：
Fisher, AG
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通讯作者：
Draper, Jonathan S.
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作者：
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