Article (Scientific journals)
Constitutive expression of tdTomato protein as a cytotoxicity and proliferation marker for space radiation biology
Chishti, Arif A.; Hellweg, Christine E.; Berger, Thomas et al.
2015In Life Sciences in Space Research, 4, p. 35-45
Peer reviewed
 

Files


Full Text
Constitutive expression.pdf
Publisher postprint (1.58 MB)
Download

All documents in ORBilu are protected by a user license.

Send to



Details



Keywords :
Cytotoxicity; Fluorescent protein; Mammalian cells; Proliferation tdTomato; Radiation response; Article; Cytomegalovirus; Mammalia
Abstract :
[en] The radiation risk assessment for long-term space missions requires knowledge on the biological effectiveness of different space radiation components, e.g. heavy ions, on the interaction of radiation and other space environmental factors such as microgravity, and on the physical and biological dose distribution in the human body. Space experiments and ground-based experiments at heavy ion accelerators require fast and reliable test systems with an easy readout for different endpoints. In order to determine the effect of different radiation qualities on cellular proliferation and the biological depth dose distribution after heavy ion exposure, a stable human cell line expressing a novel fluorescent protein was established and characterized. tdTomato, a red fluorescent protein of the new generation with fast maturation and high fluorescence intensity, was selected as reporter of cell proliferation. Human embryonic kidney (HEK/293) cells were stably transfected with a plasmid encoding tdTomato under the control of the constitutively active cytomegalovirus (CMV) promoter (ptdTomato-N1). The stably transfected cell line was named HEK-ptdTomato-N1 8. This cytotoxicity biosensor was tested by ionizing radiation (X-rays and accelerated heavy ions) exposure. As biological endpoints, the proliferation kinetics and the cell density reached 100 h after irradiation reflected by constitutive expression of the tdTomato were investigated. Both were reduced dose-dependently after radiation exposure. Finally, the cell line was used for biological weighting of heavy ions of different linear energy transfer (LET) as space-relevant radiation quality. The relative biological effectiveness of accelerated heavy ions in reducing cellular proliferation peaked at an LET of 91 keV/μm. The results of this study demonstrate that the HEK-ptdTomato-N1 reporter cell line can be used as a fast and reliable biosensor system for detection of cytotoxic damage caused by ionizing radiation. © 2015 The Committee on Space Research (COSPAR).
Disciplines :
Microbiology
Identifiers :
eid=2-s2.0-84924077535
Author, co-author :
Chishti, Arif A.;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany, Karachi Institute of Biology and Genetic Engineering (KIBGE), University of Karachi, Karachi, Pakistan
Hellweg, Christine E.;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany
Berger, Thomas;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany
Baumstark-Khan, Christa;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany
Feles, Sebastian;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany
Kätzel, Thorben ;  University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit
Reitz, Günther;  German Aerospace Center (DLR), Institute of Aerospace Medicine, Radiation Biology, Linder Höhe, Köln, Germany
External co-authors :
yes
Language :
English
Title :
Constitutive expression of tdTomato protein as a cytotoxicity and proliferation marker for space radiation biology
Publication date :
2015
Journal title :
Life Sciences in Space Research
ISSN :
2214-5524
Publisher :
Elsevier Ltd
Volume :
4
Pages :
35-45
Peer reviewed :
Peer reviewed
Available on ORBilu :
since 16 May 2016

Statistics


Number of views
56 (1 by Unilu)
Number of downloads
920 (1 by Unilu)

Scopus citations®
 
7
Scopus citations®
without self-citations
6
OpenCitations
 
6

Bibliography


Similar publications



Contact ORBilu