Naslov
Animal Cell Lines as an Alternative Tool for Toxicity Testing of Xenobiotics

Autori
Kmetič, Ivana ; Šimić, Branimir ; Kniewald, Jasna

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

Izvornik
Abstract Book of the "3rd Croatian Congress of Toxicology - CROTOX 2004", May 26-29, 2004, Plitvice Lakes, Croatia / - Zagreb : Hrvatsko toksikološko društvo, 2004

Skup
3rd Croatian Congress of Toxicology-CROTOX 2004

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

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Domaća recenzija

Ključne riječi
alternative method; animall cell line; cytotoxicity; xenobiotics

Sažetak
The safety assessment of new chemicals and pharmaceuticals and the incorporation of these data into a human risk assessment package requires a large number of expensive, regulated tests in animal species, including in some cases nonhuman primates. Currently, there are a wide range of animal replacement alternative opportunites in industrial chemical and drug development. Alternatives are techniques for reducing the number of animals used in medical and scientific research, mitigating the suffering of any animals that have to be used, and whenever possible, replacing animals by other tehniques. This approach is termed the Three Rs: Reduction, Refinement and Replacement. Alternatives include subcellular fractions, perfused organs, tissue slices, primary cell cultures and cell lines. In this study we have used animal cell lines CHO (Chinese Hamster Ovary - CHO K1) and BHK (Baby Hamster Kidney - BHK 21 C13) as an alternative tool for determining the cytotoxic effects of s-triazine herbicide atrazine and organochlorine insecticides aldrin and lindane. Cell viability and number of cells were measured by Trypane Blue exclusion method in Fuchs-Roshenthal heamocytometer and lysosomal activity measured by Neutral Red method. The production of biomass was in T-bottle, and the cells, that were separated in the early logaritmic phase of growth, were seeded on multiwell plates. The initial concentration was 2.5x10^4 cells/ml/well for the CHO cells in monolayer and 2.5x10^5 cells/ml/well for BHK cells in suspension. After 24 hours BHK cells were treated with lindane in the concentrations of 10-100 microg/ml/well and with aldrin in the concentrations of 5-125 microg/ml/well. Atrazine was added to CHO cells after 16 hours in the concentrations of 5-80 microg/ml/well. The value of IC_50 for lindane after 72 h was 35.8 microg/ml, for aldrin 11.0 microg/ml and for atrazine 50.21 microg/ml which correlates with the rate of their LD_50 values (lindane LD_50 125 mg/kg, aldrin LD_50 45-50 mg/kg, atrazine LD_50 1000-1300 mg/kg). Both cell lines CHO K1 and BHK21 C13 are acceptable as an allternative toxicity testing methods for determining cytotoxicity, that can be used as the starting dose for in vivo testing by applying the standard regression between IC_50 values and LD_50 values in the Register of Cytotoxicity.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

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