БІОЛОГІЧНА ДІЯ ЕРИТРОПОЕТИНУ У РІЗНИХ КОНЦЕНТРАЦІЯХ НА КУЛЬТУРУ КЛІТИН

Abstract

У статті описано біологічну дію різних концентрацій еритропоетину на тривалість окремих стадій розвитку культури клітин. На основі отриманих показників побудували криву росту культури клітин ендотеліосаркоми при інкубації з еритропоетином у різних концентраціях. З’ясували, що еритропоетин у всіх концентраціях сприяв швидшій адаптації клітин до нового середовища, проте не стимулював їх поділ. В статье описано биологическое действие различных концентраций эритропоэтина на продолжительность отдельных стадий развития культуры клеток. На основе полученных показателей побудовали кривую роста культуры клеток ендотелиосаркомы при инкубации с эритропоэтином в различных концентрациях. Выяснили, что эритропоэтин во всех концентрациях способствовал быстрой адаптации клеток к новой среде, однако не стимулировал их разделение. Nowadays cells cultivation in vitro becomes more and more widespread in various fields of biology, agriculture and medicine. The most essential characteristic of culture is growth, namely the study of the number of cells on the different life’s stages. Accordingly, a lot of new methods of cell’s growth parameters research tend to continually develop and improve. In addition, the study of the biologically active substances influence, such as erythropoietin, on the growth parameters of the cell culture is quite popular today. It allows to determine the biological effect of these substances, depending on their concentration, and to assess the life processes that occur in cells. Erythropoietin is a hormone, which, according to several researches, has a pleiotropic effect on physiological systems of the human body. It is shown that the erythropoietin receptors were also detected in endothelial cells, smooth and skeletal muscle, the myocardium. The purpose of the study was to determine the dependence of the individual stages duration on the development of cell culture endothelioma of the mouse from erythropoietin concentration. The cell culture of the mouse endothelium was obtained by explants separate parts of the tumor and standard cultivation procedures. To determine the effect of erythropoietin on the kinetics parameters of culture cells growth was formed four research groups: the first was incubated with erythropoietin at a concentration of 13 IU/ml, the second was 6.5 IU/ml, a third was 0.13 IU/ml and the fourth was a control group. The incubation lasted 32 days. Periodically Goryaeva’s camera was counting the number of cells in culture. On the basis of the obtained data the growth curve with subsequent analysis of its separate parts was observed. During the research procedures it was found out that the growth of culture cells was characterized by four phases: the lag phase, phase of exponential growth, stationary phase, and the degradation of culture. It was also found out that erythropoietin as a biologically active substance has not the same effect on the cell culture in varying concentrations. If the concentration is high (13 IU/ml), it has a moderate dampening effect on the growth of culture cells. Although it stimulated a rapid transfer of culture cells from the lag phase to the exponential phase compared to the control group, however, the maximum cell concentration has never reached the level of the control group. At an average concentration of (6,5 IU/ml) erythropoietin had a pronounced dampening effect. The duration of the lag phase was less than in the control group, but the duration of the exponential growth phase was the same as in the control group. The maximum concentration of cells was 3.8 times less than the control. At low concentrations (0,13 IU\ml) erythropoietin almost had not inhibitory action. It contributed to a more rapid transition of the cell cultures in exponential growth, compared to the control group, and the maximum concentration of cells reached almost the same level as in the control group. To sum up, in the all concentrations erythropoietin contributed to a more rapid cells’ adaptation to new environment (reducing the duration of lag phase), but did not stimulate cell division (increase in cell concentration), medium and high concentrations suppress it. It should be also noted that during incubation with erythropoietin in average concentrations of inhibition effect on cell division was more pronounced, compared to other concentrations.