engleski

Cytotoxic effects of atrazine and lindane on rat ovarian and uterine primary culture cells

In order to determine influence of rather common used pesticides lindane (L) or atrazine (A) on reproductive tissues at female rats, this research has been performed. Our previous results had shown that A and L in vivo are responsible for estrous cycle failure and therefore we used a primary cell culture. Primary cultures were prepared from 90 days old female Fischer rats grown under the standard laboratory conditions. For preparation of primary culture tissue slices were first incubated for 20 min at 37°C with trypsine at Glasgow MEM supplemented with 0.015 M CaCl2 and later with collagenase and trypsine for 40 min. Cells were seeded at Multiwell plates at the concentration of 5x104 cells/ml. A was added at the concentrations of 20, 40 and 60 ľg/ml while L due to its higher toxicity only 10, 20 or 30 ľg/ml. After incubation at 37°C and under the atmosphere of 95% air and 5% CO2, cytotoxicity of A or L have been determined with the application of Kenacid blue and Neutral Red coloring methods. Cytotoxic effect of A on uterine cells increased with the increased concentration of A from 9.5 to 36.7%, while for L from 24.7 to 46.7% for the selected concentrations. At the same time ovarian cells under the influence of A have shown that inhibitory effect is for 23% lower and for L is 15% higher. At the same time we have find that both colorimetric methods are acceptable for the quantitative determination of cytotoxic effects with ovarian or uterine cells in culture what can reduce the need of the direct experiments with animals. Supported by Ministry of Science and Technology Republic of Croatia, grants 058104 and 058409 and by PLIVA d.d. Keywords: cytotoxicity, pesticides, reproduction

atrazine; lindane; rat; ovary; uterus; primary cell culture

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