Hellweg, Christine E., Shinde, Vaibhav, Srinivasan, Sureshkumar Perumal, Henry, Margit, Rotshteyn, Tamara, Baumstark-Khan, Christa, Schmitz, Claudia, Feles, Sebastian, Spitta, Luis F., Hemmersbach, Ruth, Hescheler, Juergen and Sachinidis, Agapios (2020). Radiation Response of Murine Embryonic Stem Cells. Cells, 9 (7). BASEL: MDPI. ISSN 2073-4409

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Abstract

To understand the mechanisms of disturbed differentiation and development by radiation, murine CGR8 embryonic stem cells (mESCs) were exposed to ionizing radiation and differentiated by forming embryoid bodies (EBs). The colony forming ability test was applied for survival and the MTT test for viability determination after X-irradiation. Cell cycle progression was determined by flow cytometry of propidium iodide-stained cells, and DNA double strand break (DSB) induction and repair by gamma H2AX immunofluorescence. The radiosensitivity of mESCs was slightly higher compared to the murine osteoblast cell line OCT-1. The viability 72 h after X-irradiation decreased dose-dependently and was higher in the presence of leukemia inhibitory factor (LIF). Cells exposed to 2 or 7 Gy underwent a transient G2 arrest. X-irradiation induced gamma H2AX foci and they disappeared within 72 h. After 72 h of X-ray exposure, RNA was isolated and analyzed using genome-wide microarrays. The gene expression analysis revealed amongst others a regulation of developmental genes (Ada, Baz1a, Calcoco2, Htra1, Nefh, S100a6andRassf6), downregulation of genes involved in glycolysis and pyruvate metabolism whereas upregulation of genes related to the p53 signaling pathway. X-irradiated mESCs formed EBs and differentiated toward cardiomyocytes but their beating frequencies were lower compared to EBs from unirradiated cells. These results suggest that X-irradiation of mESCs deregulate genes related to the developmental process. The most significant biological processes found to be altered by X-irradiation in mESCs were the development of cardiovascular, nervous, circulatory and renal system. These results may explain the X-irradiation induced-embryonic lethality and malformations observed in animal studies.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Hellweg, Christine E.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Shinde, VaibhavUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Srinivasan, Sureshkumar PerumalUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Henry, MargitUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Rotshteyn, TamaraUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Baumstark-Khan, ChristaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schmitz, ClaudiaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Feles, SebastianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Spitta, Luis F.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hemmersbach, RuthUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hescheler, JuergenUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Sachinidis, AgapiosUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-327563
DOI: 10.3390/cells9071650
Journal or Publication Title: Cells
Volume: 9
Number: 7
Date: 2020
Publisher: MDPI
Place of Publication: BASEL
ISSN: 2073-4409
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
DNA-DAMAGE RESPONSE; IONIZING-RADIATION; HOMOLOGOUS RECOMBINATION; DIFFERENTIATION PROCESSES; SIMULATED MICROGRAVITY; H2AX PHOSPHORYLATION; GENOMIC INTEGRITY; SOMATIC-CELLS; MOUSE; EXPRESSIONMultiple languages
Cell BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/32756

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