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

J 2017

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

ŠIMARA, Pavel, Lenka TESAŘOVÁ, Daniela ŘEHÁKOVÁ, Pavel MATULA, Stanislav STEJSKAL et. al.

Základní údaje

Originální název

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

Autoři

ŠIMARA, Pavel (203 Česká republika, domácí), Lenka TESAŘOVÁ (203 Česká republika, domácí), Daniela ŘEHÁKOVÁ (203 Česká republika, domácí), Pavel MATULA (203 Česká republika, domácí), Stanislav STEJSKAL (203 Česká republika, domácí), Aleš HAMPL (203 Česká republika, domácí) a Irena KRONTORÁD KOUTNÁ (203 Česká republika, garant, domácí)

Vydání

Stem Cell Research & Therapy, Springer Nature, 2017, 1757-6512

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10601 Cell biology

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.963

Kód RIV

RIV/00216224:14330/17:00094705

Organizační jednotka

Fakulta informatiky

DOI

http://dx.doi.org/10.1186/s13287-017-0522-5

UT WoS

000396978500002

Klíčová slova anglicky

Human induced pluripotent stem cells; DNA double-strand breaks; gammaH2AX; 53BP1; Long-term in vitro culture; DNA repair

Štítky

cbia-web

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 9. 2. 2022 09:16, doc. RNDr. Pavel Matula, Ph.D.

Anotace

V originále

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.

Návaznosti

GBP302/12/G157, projekt VaV

Název: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

Investor: Grantová agentura ČR, Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii

LM2015090, projekt VaV

Název: Český národní uzel Evropské sítě infrastruktur klinického výzkumu (Akronym: CZECRIN)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CZECRIN - Český národní uzel Evropské sítě infrastruktur klinického výzkumu

NV16-31501A, projekt VaV

Název: Tkáňové inženýrství epitelů: Buňky a protokoly pro regenerativní medicínu

Citovat

ŠIMARA, Pavel, Lenka TESAŘOVÁ, Daniela ŘEHÁKOVÁ, Pavel MATULA, Stanislav STEJSKAL, Aleš HAMPL a Irena KRONTORÁD KOUTNÁ. DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing. Stem Cell Research & Therapy. Springer Nature, 2017, roč. 8, č. 73, s. 1-13. ISSN 1757-6512. Dostupné z: https://dx.doi.org/10.1186/s13287-017-0522-5.

@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 a 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, roč.~8, č.~73, s.~1-13. ISSN~1757-6512. Dostupné z: https://dx.doi.org/10.1186/s13287-017-0522-5.

Podrobný výpis o publikaci