AbstractAssessing the biological risks associated with exposure to the high-energy charged particles encountered in space is essential for the success of long-term space exploration. Although prokaryotic and eukaryotic cell models developed in our laboratory and others have advanced our understanding of many aspects of genotoxicity, in vitro models are needed to assess the risk to humans from space radiation insults. Such models must be representative of the cellular interactions present in tissues and capable of quantifying genotoxic damage. Toward this overall goal, the objectives of this study were to examine the effect of the localized microenvironment of cells, cultured as either 2-dimensional (2D) monolayers or 3-dimensional (3D) aggregates, on the rate and type of genotoxic damage resulting from exposure to Fe-charged particles, a significant portion of space radiation. We used rodent transgenic cell lines containing 50–70 copies of a LacI transgene to provide the enhanced sensitivity required to quantify mutational frequency and type in the 1100-bp LacI target as well as assessment of DNA damage to the entire 45-kbp construct. Cultured cells were exposed to high-energy Fe charged particles at Brookhaven National Laboratory’s Alternating Gradient Synchrotron facility for a total dose ranging from 0.1 to 2Gy and allowed to recover for 0–7days, after which mutational type and frequency were evaluated. The mutational frequency was found to be higher in 3D samples than in 2D samples at all radiation doses. Mutational frequency also was higher at 7days after irradiation than immediately after exposure. DNA sequencing of the mutant targets revealed that deletional mutations contributed an increasingly high percentage (up to 27%) of all mutations in cells as the dose was increased from 0.5 to 2Gy. Several mutants also showed large and complex deletions in multiple locations within the LacI target. However, no differences in mutational type were found between the 2D and the 3D samples. These 3D tissue-like model systems can reduce the uncertainty involved in extrapolating risk between in vitro cellular and in vivo models.


    Access

    Check access

    Check availability in my library

    Order at Subito €


    Export, share and cite



    Title :

    Comparison of genotoxic damage in monolayer cell cultures and three-dimensional tissue-like cell assemblies


    Contributors:
    Behravesh, E. (author) / Emami, K. (author) / Wu, H. (author) / Gonda, S. (author)

    Published in:

    Advances in Space Research ; 35 , 2 ; 260-267


    Publication date :

    2005-01-24


    Size :

    8 pages




    Type of media :

    Article (Journal)


    Type of material :

    Electronic Resource


    Language :

    English





    Comparison of genotoxic damage in monolayer cell cultures and three-dimensional tissue-like cell assemblies

    Behravesh, E. / Emami, K. / Wu, H. et al. | British Library Conference Proceedings | 2005



    Growing Three-Dimensional Cartilage-Cell Cultures

    Spaulding, Glenn F. / Prewett, Tacey L. / Goodwin, Thomas J. | NTRS | 1995


    Three-Dimensional Transgenic Cell Model to Quantify Genotoxic Effects of Space Environment

    Gonda, S. R. / Wu, H. / Pingerelli, P. L. et al. | British Library Conference Proceedings | 2001