DNA double strand breaks detection in pluripotent stem cells with regard to cell cycle stages

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14330%2F16%3A00088039" target="_blank" >RIV/00216224:14330/16:00088039 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

DNA double strand breaks detection in pluripotent stem cells with regard to cell cycle stages

Popis výsledku v původním jazyce

Human induced pluripotent stem cells (hiPSCs) have a potential in both disease modeling and regenerative medicine. It is of utmost importance that genomic integrity of the cells remains unharmed and DNA reparation systems fully functional. In our research we focus on the detection of DNA double strand breaks (DSBs) by phosphorylated histone H2AX (known as gammaH2AX) and p53-binding protein 1 (53BP1) in fibroblasts, three distinct lines of hiPSCs, and one line of human embryonic stem cells (hESCs). We measured both spontaneously occurred DSBs and DSBs induced by gamma-irradiation and its decrease in time. Foci number was detected by fluorescence microscopy and EdU (5-ethynyl-2-deoxyuridine) was used to discriminate between cell cycle stages. Discrimination between the EdU negative (G1) and positive (S/G2) populations allows excluding the replication-related foci and increase the accuracy of measurement. This is crucial when comparing number of DSBs in cell types with different cell cycle speed (ie.

Název v anglickém jazyce

DNA double strand breaks detection in pluripotent stem cells with regard to cell cycle stages

Popis výsledku anglicky

Human induced pluripotent stem cells (hiPSCs) have a potential in both disease modeling and regenerative medicine. It is of utmost importance that genomic integrity of the cells remains unharmed and DNA reparation systems fully functional. In our research we focus on the detection of DNA double strand breaks (DSBs) by phosphorylated histone H2AX (known as gammaH2AX) and p53-binding protein 1 (53BP1) in fibroblasts, three distinct lines of hiPSCs, and one line of human embryonic stem cells (hESCs). We measured both spontaneously occurred DSBs and DSBs induced by gamma-irradiation and its decrease in time. Foci number was detected by fluorescence microscopy and EdU (5-ethynyl-2-deoxyuridine) was used to discriminate between cell cycle stages. Discrimination between the EdU negative (G1) and positive (S/G2) populations allows excluding the replication-related foci and increase the accuracy of measurement. This is crucial when comparing number of DSBs in cell types with different cell cycle speed (ie.

Druh

O - Ostatní výsledky

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

<a href="/cs/project/GBP302%2F12%2FG157" target="_blank" >GBP302/12/G157: Dynamika a organizace chromosomů během buněčného cyklu a při diferenciaci v normě a patologii</a><br>

Návaznosti

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

Rok uplatnění

2016

Kód důvěrnosti údajů

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