engleski

Application of Chinese Hamster Ovary (CHO K1) cell line as an alternative toxicity testing of insecticide lindane

Alternatives are techniques for reducing the number of animals used in medical and scientific research, mitigating the suffering of any animal that have to be used, and whenever possible, replacing animals by other techniques. Cell lines are widely used because they do not require the use of fresh tissue. In the present study we have used animal cell line CHO (Chinese Hamster Ovary - CHO K1) in monolayer in order to evaluate the effects of insecticide lindane (gamma-hexachlorocyclohexane) on reproduction and to reduce the expensive and regulated tests usually used for the safety assessment of pesticides. Cell viability and number of cells were measured by Trypane Blue exclusion method, lyzosomal activity by Neutral Red method and changes in total cell proteins by Kenacid Blue R binding method. Cells were maintained at 37 ^0C in an atmosphere of 95% air and 5% CO_2 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. Initial concentrations of CHO cells were 1x10^4 cells/mL/well on 6-multiwell plates. Cells were seeded 16 h before the treatment with lindane (10, 20, 30, 50, 70 or 100 microg/mL/well). Cytotoxicity was determined after 24, 48 and 72 hours and was dose-responded. IC_50 values were calculated from plots of % inhibition vs. log concentrations of lindane after 72 hours of exposure for each applied method, that can be used as the starting dose for in vivo testing by applying the standard regression between IC_50 and LD_50 values in the Register of Cytotoxicity.

CHO K1 cell line; cytotoxicity; lindane

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