DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14330%2F17%3A00094705" target="_blank" >RIV/00216224:14330/17:00094705 - isvavai.cz</a>

Result on the web

<a href="https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0522-5" target="_blank" >https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0522-5</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1186/s13287-017-0522-5" target="_blank" >10.1186/s13287-017-0522-5</a>

Result language

angličtina

Original language name

DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing

Original language description

BACKGROUND: Human induced pluripotent stem cells (hiPSCs) play roles in both disease modelling and regenerative medicine. It is critical that the genomic integrity of the cells remains intact and that the DNA repair systems are fully functional. In this article, we focused on the detection of DNA double-strand breaks (DSBs) by phosphorylated histone H2AX (known as gammaH2AX) and p53-binding protein 1 (53BP1) in three distinct lines of hiPSCs, their source cells, and one line of human embryonic stem cells (hESCs). METHODS: We measured spontaneously occurring DSBs throughout the process of fibroblast reprogramming and during long-term in vitro culturing. To assess the variations in the functionality of the DNA repair system among the samples, the number of DSBs induced by gamma-irradiation and the decrease over time was analysed. The foci number was detected by fluorescence microscopy separately for the G1 and S/G2 cell cycle phases. RESULTS: We demonstrated that fibroblasts contained a low number of non-replication-related DSBs, while this number increased after reprogramming into hiPSCs and then decreased again after long-term in vitro passaging. The artificial induction of DSBs revealed that the repair mechanisms function well in the source cells and hiPSCs at low passages, but fail to recognize a substantial proportion of DSBs at high passages. CONCLUSIONS: Our observations suggest that cellular reprogramming increases the DSB number but that the repair mechanism functions well. However, after prolonged in vitro culturing of hiPSCs, the repair capacity decreases.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10601 - Cell biology

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Stem Cell Research & Therapy

ISSN

1757-6512

e-ISSN

—

Volume of the periodical

8

Issue of the periodical within the volume

73

Country of publishing house

DE - GERMANY

Number of pages

13

Pages from-to

1-13

UT code for WoS article

000396978500002

EID of the result in the Scopus database

2-s2.0-85016053500