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Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants
In this work, we shed new light on the highly debated issue of chromatin fragmentation in cryopreserved cells. Moreover, for the first time, we describe replicating cell-specific DNA damage and higher-order chromatin alterations after freezing and thawing. We identified DNA structural changes associ...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6168476/ https://www.ncbi.nlm.nih.gov/pubmed/30279538 http://dx.doi.org/10.1038/s41598-018-32939-5 |
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author | Falk, Martin Falková, Iva Kopečná, Olga Bačíková, Alena Pagáčová, Eva Šimek, Daniel Golan, Martin Kozubek, Stanislav Pekarová, Michaela Follett, Shelby E. Klejdus, Bořivoj Elliott, K. Wade Varga, Krisztina Teplá, Olga Kratochvílová, Irena |
author_facet | Falk, Martin Falková, Iva Kopečná, Olga Bačíková, Alena Pagáčová, Eva Šimek, Daniel Golan, Martin Kozubek, Stanislav Pekarová, Michaela Follett, Shelby E. Klejdus, Bořivoj Elliott, K. Wade Varga, Krisztina Teplá, Olga Kratochvílová, Irena |
author_sort | Falk, Martin |
collection | PubMed |
description | In this work, we shed new light on the highly debated issue of chromatin fragmentation in cryopreserved cells. Moreover, for the first time, we describe replicating cell-specific DNA damage and higher-order chromatin alterations after freezing and thawing. We identified DNA structural changes associated with the freeze-thaw process and correlated them with the viability of frozen and thawed cells. We simultaneously evaluated DNA defects and the higher-order chromatin structure of frozen and thawed cells with and without cryoprotectant treatment. We found that in replicating (S phase) cells, DNA was preferentially damaged by replication fork collapse, potentially leading to DNA double strand breaks (DSBs), which represent an important source of both genome instability and defects in epigenome maintenance. This induction of DNA defects by the freeze-thaw process was not prevented by any cryoprotectant studied. Both in replicating and non-replicating cells, freezing and thawing altered the chromatin structure in a cryoprotectant-dependent manner. Interestingly, cells with condensed chromatin, which was strongly stimulated by dimethyl sulfoxide (DMSO) prior to freezing had the highest rate of survival after thawing. Our results will facilitate the design of compounds and procedures to decrease injury to cryopreserved cells. |
format | Online Article Text |
id | pubmed-6168476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-61684762018-10-05 Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants Falk, Martin Falková, Iva Kopečná, Olga Bačíková, Alena Pagáčová, Eva Šimek, Daniel Golan, Martin Kozubek, Stanislav Pekarová, Michaela Follett, Shelby E. Klejdus, Bořivoj Elliott, K. Wade Varga, Krisztina Teplá, Olga Kratochvílová, Irena Sci Rep Article In this work, we shed new light on the highly debated issue of chromatin fragmentation in cryopreserved cells. Moreover, for the first time, we describe replicating cell-specific DNA damage and higher-order chromatin alterations after freezing and thawing. We identified DNA structural changes associated with the freeze-thaw process and correlated them with the viability of frozen and thawed cells. We simultaneously evaluated DNA defects and the higher-order chromatin structure of frozen and thawed cells with and without cryoprotectant treatment. We found that in replicating (S phase) cells, DNA was preferentially damaged by replication fork collapse, potentially leading to DNA double strand breaks (DSBs), which represent an important source of both genome instability and defects in epigenome maintenance. This induction of DNA defects by the freeze-thaw process was not prevented by any cryoprotectant studied. Both in replicating and non-replicating cells, freezing and thawing altered the chromatin structure in a cryoprotectant-dependent manner. Interestingly, cells with condensed chromatin, which was strongly stimulated by dimethyl sulfoxide (DMSO) prior to freezing had the highest rate of survival after thawing. Our results will facilitate the design of compounds and procedures to decrease injury to cryopreserved cells. Nature Publishing Group UK 2018-10-02 /pmc/articles/PMC6168476/ /pubmed/30279538 http://dx.doi.org/10.1038/s41598-018-32939-5 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Falk, Martin Falková, Iva Kopečná, Olga Bačíková, Alena Pagáčová, Eva Šimek, Daniel Golan, Martin Kozubek, Stanislav Pekarová, Michaela Follett, Shelby E. Klejdus, Bořivoj Elliott, K. Wade Varga, Krisztina Teplá, Olga Kratochvílová, Irena Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title | Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title_full | Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title_fullStr | Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title_full_unstemmed | Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title_short | Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
title_sort | chromatin architecture changes and dna replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6168476/ https://www.ncbi.nlm.nih.gov/pubmed/30279538 http://dx.doi.org/10.1038/s41598-018-32939-5 |
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