Assessing the potential of fish cell lines as tools for the cytotoxicity testing of estuarine sediment aqueous elutriates.
In the present study, we assess the potential of fish cell lines (CHSE, EPC and RTG-2) to be used as screening tools for the ecotoxicological assessment of estuarine sediments. The processing of sediment to a form suitable for in vitro exposure is an inherent problem when using cell cultures. The approach employed in this study was to prepare aqueous elutriate extracts from whole sediments, which were subsequently used to reconstitute powdered media. This procedure allowed the exposure of cell cultures to concentrations of up to and including 100% of the original aqueous sample. Cytotoxicity was assessed using multiple endpoint measurements. Cell viability was quantified using the neutral red and alamar blue colorimetric assays, which specifically assess lysosomal and mitochondrial function, respectively. In addition, the total protein content of the cells was measured using the coomassie blue assay. Initial tests were conducted to ensure that any resultant cytotoxicity was due to sample contaminants and not osmotic stress. In addition, elutriate samples were spiked with a model toxicant to verify the ability of the cell lines to detect and respond to bioavailable contaminants. Chemical analyses were conducted on sediments from all sampling sites to assist in interpreting any observed cytotoxicity. A differential response was observed for the cytotoxicity assays following exposure treatments, which emphasises the importance of employing multiple endpoints for the determination of toxicity. Of the three cell lines utilised in this study, RTG-2 cells were the most suitable for the testing of estuarine aqueous elutriate samples on the basis of tolerance to osmolality effects. Slight toxicity was observed following exposure to the aqueous elutriates tested in this study using RTG-2 cells and the alamar blue assay. In order to fully evaluate the overall sensitivity of this cell line, further research is warranted using an extensive range of test sites incorporating more polluted sediments.
Davoren
M
M
Radiation and Environmental Science Centre, FOCAS Institute, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland. maria.davoren@dit.ie
Ní Shúilleabháin
S
S
Hartl
M G J
MG
Sheehan
D
D
O'Brien
N M
NM
O'Halloran
J
J
Van Pelt
F N A M
FN
Mothersill
C
C
D000826:Animal Testing Alternatives; D000818:Animals; D002347:Carps; D002460:Cell Line / Q000187:drug effects* / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005399:Fishes; D019015:Geologic Sediments / Q000737:chemistry*; D009499:Neutral Red / Q000378:metabolism; D017686:Oncorhynchus mykiss; D010078:Oxazines / Q000378:metabolism; D012474:Salmon; D018675:Toxicity Tests / Q000379:methods*; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D014966:Xanthenes / Q000378:metabolism
D010078:Oxazines; D014874:Water Pollutants, Chemical; D014966:Xanthenes; C005843:resazurin; D009499:Neutral Red