A.A. Oleshkevich - Ph.D.(Biol.), Associate Professor, Department of Informational Technologies, Mathematics and Physics, Moscow State Veterinary Academy
E.V. Kaminskaya - Senior Research Scientist, Institute of Medical-Biological Problems Russian Academy of Sciences
A.M. Nosovskiy - Dr.Sc.(Biol.), Leading Scientific. Employee, Institute of Medical-Biological Problems Russian Academy of Sciences

The optimal concentrations of Con A, PHA, EDTA, and yeast extract PABA to stimulate the MDBK cell culture proliferation were obtained. The largest increase MDBK cell mass resulted when cells were treated with Con A and PHA. Con A at the optimum concentration of 1.0 mg/ml promoted increase in the proliferation rate and "crop" cells to 1.1-106per ml at an initial concentration landing 8.0-104 cells in 1 ml of cells and the number of control 3.1 - 105 per ml. PHA in the optimal concentration of 0.01 mg/ml increased cell mass up to 9,6-105 in 1 ml. The stimulatory EDTA effect was observed when cells were treated by 0.01 mg/ml concentration. This contributed to improvement similar proliferate activity, as well as when making PHA, the proliferate MDBK cells index 3 times increased in compare to the control. PABA concentration in the experiments varied in the range 0.001-1 mg/ml. The greatest stimulating effect on the growth had been shown by PABA at 0.01 mg/ml. Yeast extract, rich in vitamins, amino acids, also had a significant stimulatory effect on the growth of transplantable cell culture MDBK. Growth of cells at the optimal concentration of the yeast extract of 0.1 mg/ml was 8-105 cells/ml. Proliferation index increased by 1.75 times (from 5.75 to 10). The effect of ultrasound on the rate of cell proliferation. The stimulating effect upon the treatment of low high-frequency US intensities set within 0.03-0.05 W/cm2 exposure of 5-30s. Growth of cell mass compared with control was 65-130 %, depending on the duration of exposure. The maximum stimulating effect on the ultrasonic treatment of cells has been set to 0.05 W/cm2 exposure 10s. Higher intensity of irradiation (0.1 W/cm2) resulted in a significant reduction in growth of cells and their complete destruction when exposed to US system 0.2 W/cm2. With increasing exposure time and increasing intensity US reduced the number of living, intact cells. An increasing number of bits of cells in the medium changes the morphology of cells: cells are greatly reduced in size, there are processes of the cytoplasmatic membrane, it may be a gap of cell membranes, nuclei and mitochondria , as well as a complete degeneration of the cells. Plasmolemma destruction and nuclear membrane rupture occurred due to the occurrence of microflows inside cells that accompanies the ultrasonic cavitations in biological media. Maximum growth in control cells was 3,1-105 in 1 ml. Proliferation index increased as a result of US- stimulating growth rate from 3.8 to 9.0. Microscopic inspection revealed no morphological changes in MDBK cell culture( ultrasonic intensity 0.01-0.05 W-cm-2, 5s- 1 min).Thus, ultrasonic stimulation of growth and proliferation of MDBK cell culture allowed to increase growth and reduce the time for obtaining a unit of cell mass. The combined action of US and EDTA were tested (previously selected optimum US intensity 0.05 W/cm2, exposure time 5, 10, 30s), and the concentration of EDTA 10-4-10-1 mg/ml. However, mentioned above combined effect reduced the proliferation in comparison with the action of one of these factors. The largest harvest gave MDBK cells - 5.5-105/ml. Joint use of US and mutagen leads to increased mutagenic effect by changing the polarization membrane. The lower concentrations of EDTA in the combined exposure are productive, so EDTA concentrations required for stimulation of cellular proliferation were tenfold reduced (the increase of cells was 5.5-105/ml; 103dilution of stimulator). US with Con A and PHA increased the rate of proliferation of MDBK cell culture similarly. The data show that, after cell treatment with US their need in Con A and PHA as stimulators of growth is reduced more than tenfold. It is interesting to note that when cells were treated with both US and Con A, the concentration of Con A, which stimulates the proliferate activity of the MDBK cell culture, decreased 1000-fold, compared with the concentration of Con A, which stimulates the proliferation individually. Similar results were obtained with the combined action of PHA and US in optimal intensity of 0.05 W-cm-2, 10s. Stimulative PHA concentration decreased 10-fold. The number of cells increased 3-fold compared with the control. In the case of adding EDTA to the optimally "insonated" cell suspension, the cell proliferation stimulating concentration of EDTA was 10 times lower, but the number of grown cells was only 2-fold greater than the control.