DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion

被引：61

作者：

Tsubouchi, Tomomi ^{[1
,2
]}

Soza-Ried, Jorge ^{[1
]}

Brown, Karen ^{[1
]}

Piccolo, Francesco M. ^{[1
]}

Cantone, Irene ^{[1
]}

Landeira, David ^{[1
]}

Bagci, Hakan ^{[1
]}

Hochegger, Helfrid ^{[2
]}

Merkenschlager, Matthias ^{[1
]}

Fisher, Amanda G. ^{[1
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, MRC Clin Sci Ctr, Lymphocyte Dev Grp, London W12 0NN, England

[2] Univ Sussex, MRC Genome Damage & Stabil Ctr, Brighton BN1 9RQ, E Sussex, England

来源：

CELL | 2013年 / 152卷 / 04期

基金：

欧洲研究理事会; 英国医学研究理事会;

关键词：

SOMATIC-CELLS; HYBRID-CELLS; G1; PHASE; IN-VITRO; PLURIPOTENT CELLS; CYCLE REGULATION; GENE-EXPRESSION; SELF-RENEWAL; ES CELLS; REPLICATION;

D O I：

10.1016/j.cell.2013.01.012

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

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.

引用

页码：873 / 883

页数：11

共 73 条

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