DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion

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...

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Main Authors:	Tsubouchi, Tomomi, Soza-Ried, Jorge, Brown, Karen, Piccolo, Francesco M., Cantone, Irene, Landeira, David, Bagci, Hakan, Hochegger, Helfrid, Merkenschlager, Matthias, Fisher, Amanda G.
Format:	Online
Language:	English
Published:	Cell Press 2013
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3605571/

id	pubmed-3605571
recordtype	oai_dc
spelling	pubmed-36055712013-03-22 DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion Tsubouchi, Tomomi Soza-Ried, Jorge Brown, Karen Piccolo, Francesco M. Cantone, Irene Landeira, David Bagci, Hakan Hochegger, Helfrid Merkenschlager, Matthias Fisher, Amanda G. Article 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. Cell Press 2013-02-14 /pmc/articles/PMC3605571/ /pubmed/23415233 http://dx.doi.org/10.1016/j.cell.2013.01.012 Text en © 2013 ELL & Excerpta Medica. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Tsubouchi, Tomomi Soza-Ried, Jorge Brown, Karen Piccolo, Francesco M. Cantone, Irene Landeira, David Bagci, Hakan Hochegger, Helfrid Merkenschlager, Matthias Fisher, Amanda G.
spellingShingle	Tsubouchi, Tomomi Soza-Ried, Jorge Brown, Karen Piccolo, Francesco M. Cantone, Irene Landeira, David Bagci, Hakan Hochegger, Helfrid Merkenschlager, Matthias Fisher, Amanda G. DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
author_facet	Tsubouchi, Tomomi Soza-Ried, Jorge Brown, Karen Piccolo, Francesco M. Cantone, Irene Landeira, David Bagci, Hakan Hochegger, Helfrid Merkenschlager, Matthias Fisher, Amanda G.
author_sort	Tsubouchi, Tomomi
title	DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
title_short	DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
title_full	DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
title_fullStr	DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
title_full_unstemmed	DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion
title_sort	dna synthesis is required for reprogramming mediated by stem cell fusion
description	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.
publisher	Cell Press
publishDate	2013
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3605571/
_version_	1611964676543873024

DNA Synthesis Is Required for Reprogramming Mediated by Stem Cell Fusion

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