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(PP252) Identification of radiation responsive biomarkers using a rotary bioreactor to simulate an in vivo environment.
Qutob, Sami*,1, Lachapelle, Sylvie2, Thorleifson, Erika1, Segura, Tamika2, Mullins, Dana2, Prud'homme-Lalonde, Louise, Wilkinson, Diana2, 1 Consumer and Clinical Radiation Protection Bureau, Ottawa, Ontario, Canada2 Radiological Analysis and Defence, Ottawa, Ontario, Canada
ABSTRACT- Evidence from literature identifies a number of potential radiation responsive proteins in blood. However, acquisition of samples from exposed individuals for the determination of these biomarkers that correlate with the incurred actual dosage is difficult. Therefore, the use of a tissue culture system that could simulate the in vivo environment for the purpose of biomonitoring, by identifying biomarkers of radiation exposure, would be ideal. We have found that a rotary bioreactor system provided us with superior culture conditions for the human lymphoblastoid cell line (TK-6) in comparison to the standard cell culture method. Prior to incubation, in both systems, cells were given a 2 Gy dose of gamma-radiation or were not irradiated. The endpoints measured included changes in pH, cell growth, viability, and glucose depletion. Of the nine cytokines that were examined (IL-1b, IL-2, IL-6, IL-8, IL-10, IL-12, G-CSF, GM-CSF and TNF-alpha) we identified TNF-alpha, GM-CSF, and IL-6 as possible markers of radiation exposure in this cell line. The pattern of expression of these cytokines that were present in both culture systems may be indicative of radiation exposure and could also give additional information on the amount of time that had elapsed since exposure. Future studies with different cell types and/or whole tissue samples will also be used enabling a more complete profile of protein responsiveness. The knowledge gained will assist in the development of a prototype deployable assay and provide further insight into the bystander effect.
Key words: bioreactor, cytokines, biomarkers, lymphoblastoid
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