Naslov
Cytotoxicity Testing of Atrazine and Lindane in Animal Cell Line (Chinese Hamster Ovary - CHO K1)

Autori
Kmetič, Ivana ; Janušić, Vanja ; Kniewald, Jasna

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstract Book / - Zagreb : Hrvatsko toksikološko društvo, 2004, 87-87

Skup
3^rd Croatian Congress of Toxicology-CROTOX 2004

Mjesto i datum
Plitvička Jezera, Hrvatska, 26.05.2004. - 29.05.2004

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
lindane; atrazine; CHO K1 cell line; cytotoxicity

Sažetak
Massive amounts of pesticides are produced and there are growing evidences that they are able to disturb the biological functions of non-target organisms, including humans. Atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) is a widely used agricultural herbicide, even declared to be a low toxic compound, and lindane (c-hexachlorocyclohexane) is an insecticide with the restricted permission of usage, but their residues in the environment may present a risk for the reproductive processes. Ovary cell culture line from chinese hamster were introduced, in order to reduce the expensive and regulated tests in different animal species usually used for the safety assessment of pesticides. In the present study, in order to determine cytotoxic effect of atrazine and lindane within the reproductive hormone-dependent tissues, using CHO K1 (Chinese Hamster Ovary) cell line in monolayer (CCL-61) performed the experiments. Cell viability and number of cells were measured by Trypane Blue exclusion method in Fuchs-Rosenthal heamocytometer, lyzosomal activity were checked by Neutral Red method and change in total cell protein by Kenacid Blue R binding method. The cells were maintained at 370C in an atmosphere of 95% air and 5% CO2 in Dulbecco medium supplemented with 10% fetal calf serum. The production of biomass was in T-bottle, and the cells were separated in the early logarithmic phase of growth. The initial concentrations of CHO cells were 1x10^4 cells/mL/well on 6-multiwell plates. Cells were seeded 16 h before the treatment with atrazine (20, 40, 60 or 80 microg/mL/well) or lindane (10, 20, 30, 50, 70 or 100 microg/mL/well). The cytotoxicity was determined after 24, 48 and 72 hours. The cell growth inhibition ranged after 72 h was dose-responded. IC_50 values were calculated from plots of % inhibition vs. log concentrations of tested compounds after 72 hours of exposure. Determined IC_50 values for atrazine and lindane after 72 hours of exposure enable the first-step in toxicity evaluation of such xenobiotics. By excluding the experiments with animals, application of cell lines from reproductive tissues like the Chinese Hamster Ovary, enables evaluation of toxicity range in the reproductive tissues.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

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