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@article{1377715,
author = {Šimara, Pavel and Tesařová, Lenka and Řeháková, Daniela and Matula, Pavel and Stejskal, Stanislav and Hampl, Aleš and Krontorád Koutná, Irena},
article_number = {73},
doi = {http://dx.doi.org/10.1186/s13287-017-0522-5},
keywords = {Human induced pluripotent stem cells; DNA double-strand breaks; gammaH2AX; 53BP1; Long-term in vitro culture; DNA repair},
language = {eng},
issn = {1757-6512},
journal = {Stem Cell Research & Therapy},
title = {DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing},
url = {https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0522-5},
volume = {8},
year = {2017}
}
TY - JOUR
ID - 1377715
AU - Šimara, Pavel - Tesařová, Lenka - Řeháková, Daniela - Matula, Pavel - Stejskal, Stanislav - Hampl, Aleš - Krontorád Koutná, Irena
PY - 2017
TI - DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing
JF - Stem Cell Research & Therapy
VL - 8
IS - 73
SP - 1-13
EP - 1-13
PB - Springer Nature
SN - 17576512
KW - Human induced pluripotent stem cells
KW - DNA double-strand breaks
KW - gammaH2AX
KW - 53BP1
KW - Long-term in vitro culture
KW - DNA repair
UR - https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0522-5
L2 - https://stemcellres.biomedcentral.com/articles/10.1186/s13287-017-0522-5
N2 - 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.
ER -
ŠIMARA, Pavel, Lenka TESAŘOVÁ, Daniela ŘEHÁKOVÁ, Pavel MATULA, Stanislav STEJSKAL, Aleš HAMPL and Irena KRONTORÁD KOUTNÁ. DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing. \textit{Stem Cell Research \&{} Therapy}. Springer Nature, 2017, vol.~8, No~73, p.~1-13. ISSN~1757-6512. Available from: https://dx.doi.org/10.1186/s13287-017-0522-5.
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