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Influence of ionizing radiation on proliferation of murine cells and cytotoxicity of EdU revealed by dual pulse and chase analysis

Teber, Denise (2015) Influence of ionizing radiation on proliferation of murine cells and cytotoxicity of EdU revealed by dual pulse and chase analysis. Master's, University of Cologne.

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Abstract

Astronauts are constantly exposed to different irradiation doses, resulting in distinct short- and long-term consequences for the organism. Impaired cell cycle progression and decreased clonogenic survival can be caused by elevated radiation levels. To fully understand the effects of different radiation types and doses, a constant development of new methods is indispensable. In order to determine the current cell cycle phase the DNA content can be visualized by DNA intercalating agents and for the analysis of cellular proliferation thymidine analogs can be incorporated into newly synthesized DNA. To measure the effects of irradiation on cellular proliferation and on cell cycle progression in one experiment, a new approach is used by combining two methods. The new assay is called dual pulse labeling. Two different thymidine analogs were used in combination to allow labeling at two different time points, followed by the staining of the DNA. Impaired proliferation of MEF NEMO KO cells was observed after X-irradiation. Irradiation with silicon ions lead to an increased number of MEF in the G2/M phase of cell cycle. Furthermore, cytotoxic side effects were determined for the application of one thymidine analog (5-Ethynyl-2´-deoxyuridine) in mouse embryonic fibroblasts. The results of this study facilitate a new possibility to assess the consequences of radiation on the cell cycle progression and cellular proliferation in one experiment, using fluorescence microscopy and flow cytometry as a read out. Furthermore, the results show that the new thymidine analog EdU can be useful in many scientific questions, but it is inevitable to check for the suitability of the used cell line and to keep the time frame of the experiment restricted.

Item URL in elib:https://elib.dlr.de/98239/
Document Type:Thesis (Master's)
Title:Influence of ionizing radiation on proliferation of murine cells and cytotoxicity of EdU revealed by dual pulse and chase analysis
Authors:
AuthorsInstitution or Email of AuthorsAuthors ORCID iD
Teber, DeniseRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany.UNSPECIFIED
Date:2015
Refereed publication:Yes
Open Access:No
Gold Open Access:No
In SCOPUS:No
In ISI Web of Science:No
Number of Pages:81
Status:Published
Keywords:ionizing radiation, dual pulse labeling, DNA
Institution:University of Cologne
Department:Department for Biology
HGF - Research field:Aeronautics, Space and Transport
HGF - Program:Space
HGF - Program Themes:Research under Space Conditions
DLR - Research area:Raumfahrt
DLR - Program:R FR - Forschung unter Weltraumbedingungen
DLR - Research theme (Project):R - Vorhaben Strahlenbiologie
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Radiation Biology
Deposited By: Kopp, Kerstin
Deposited On:17 Sep 2015 10:54
Last Modified:17 Sep 2015 10:54

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