Clamping down on copy errors

Kad, Neil M.; Houten, Bennett Van

doi:10.1038/nature20475

Cited by 1 publication

(1 citation statement)

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“…In the present study, we correlate cell viability with freeze-thawed DNA integrity and chromatin states as explored by high-resolution confocal fluorescence microscopy and flow cytometry 19 – 23 , and we are the first to identify novel critical attributes of chromatin damage, shedding new light on the mechanisms of freeze-thaw-induced chromatin alteration, consequent cell survival, and cryoprotection. DNA double strand breaks (DSBs) represent the most serious DNA lesions 20 , 21 , 24 , 25 , but their induction through the freeze-thaw process remains controversial 26 – 29 . We have shown that freezing and thawing preferentially damage replicating (S-phase) cells by causing the collapse of replication forks, eventually leading to DSBs, thereby making rapidly dividing cells more sensitive to freeze damage.…”

Section: Introductionmentioning

confidence: 99%

Chromatin architecture changes and DNA replication fork collapse are critical features in cryopreserved cells that are differentially controlled by cryoprotectants

Falk

Falková

Kopečná

et al. 2018

Sci Rep

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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.

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