О. В. Штапенко, Ю. И. Слывчук, И. И. Гевкан

Институт биологии животных НААН, Львов

ВПЛИЯНИЕ ХЛОРИДА НИКЕЛЯ НА МОРФОФУНКЦИОНАЛЬНЫЕ И МЕТАБОЛИЧЕСКИЕ ХАРАКТЕРИСТИКИ КЛЕТОК IN VITRO

Исследована зависимость роста и метаболических изменений в клетках яйцеводов коров при различной концентрации хлорида никеля. Показано, что обе концентрации хлорида никеля 100 и 150 мкг/мл вызывают снижение пролиферативного роста клеток яйцеводов в течение 72 часов культивирования, однако более выраженное влияние выявлено при более высокой дозе. Снижение интенсивности пролиферации клеток, обусловленное хлоридом никеля сопровождалось изменениями метаболических процессов в культуре клеток. Выявлено, что высокая концентрация достоверно снижает выживаемость клеток, их способность к делению (р<0.001), снижает интенсивность потребления фосфора (р<0.001) и достоверно повышает содержание кальция (р<0.01).

Ключевые слова: культура клеток in vitro, никель хлорид, пролиферация, цитотоксичность

O. V. Shtapenko, Yu. I. Slyvchuk, I. I. Gevkan/h3>

Institute of Animal Biology NAAS, Lviv, Ukraine

INFLUENCE OF NICKEL CHLORIDE ON MORPHOFUNCTIONAL AND METABOLIC CHARACTERISTICS OF CELLS IN VITRO

Increasing pollution of the environment by heavy metal compounds have harmful effects on living organisms, they could lead to disorders of many physiological functions, in particular, reproductive system. Nickel is a cofactor of some cellular enzymes, Ni2+ ions stabilize the structure of nucleic acids and ribosomes, however at the high concentrations he become noxious. The dependence of growth and metabolic changes in the cells of the oviducts of cows at different concentrations of nickel chloride was evaluated. Research was performed on a primery culture of ovine oviduct epithelial cells (BOECs). For experiments, BOECs were seeded into a 35-mm Petri dish at 1.2×106 cells per dish in TCM-199 and 40 µg/mL gentamicine supplemented with 10% of fetal bovine serum. Water soluble salt of nickel chloride was added to the cell culture at concentrations of 100 and 150 µg/ml, 24 hours after the passing. Using the cellular viability parameters (cell growth and proliferation) and metabolic cell parameters (concentration of total protein, glucose levels, calcium and phosphorus content) the vitality of BOECs in vitro after treatment with nickel chloride over 72 hours incubation periods was studied. Experimental studies demonstrated that proliferation activity oviduct cells treated with nickel chloride decreased under 72 h over controls, however addition of 150 µg/ml nickel chloride lead to more destructive changes in a cell culture. The decrease the intensity of cell proliferation by the influence of nickel chloride is confirmed by changes of metabolic processes in culture of oviduct cells. At 48 and 72 hours, the content of total protein a statistically (Р<0,01; Р<0,001) decreased in both experimental groups compared to the control. It was found that the higher concentration significantly decreases the survival of cells, their ability to divide (P<0,001), reduces the intensity of phosphorus consumption (P<0.001) and significantly increases the calcium content (P<0,01). A decrease in the glucose content in all stages of the cellular cultivation for control and both experimental groups was observed. These results show that nickel chloride in experimental concentrations have no cytotoxic effect on BOECs because the evaluation of glucose uptake is crucial in the study of cell viability and numerous metabolic disorders. Experimental studies demonstrated time-dependent effect of nickel chloride on the BOECs. It was shown that both concentrations of nickel chloride 100 and 150 μg/ml caused a decrease in the proliferative growth of bovine oviduct cells during 72 hours of cultivation, however, a more pronounced effect was detected at a higher dose.

Keywords: in vitro cell culture, nickel chloride, proliferative activity, cytotoxicity