R.K. Chailakhyan - Dr.Sc. (Med.), Professor, Head of Laboratory of Stromal Immune Regulation, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei of RAMS, Moscow, Russia. E-mail: rubenchail@yandex.ru Y.V. Gerasimov - Ph.D. (Biol.), Senior Research Scientist, Head of Laboratory of Stromal Immune Regulation, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei of RAMS, Moscow, Russia V.G. Lunin - Dr.Sc. (Biol.), Head of Laboratory of Bioactive Nanostructures, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei, RAMS, Moscow, Russia V.I. Korobko - Research Scientist, Laboratory of Bioactive Nanostructures, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei, RAMS, Moscow, Russia A.I. Kuralesova - Ph.D. (Biol.), Leading Research Scientist, Laboratory of Stromal Immune Regulation, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei of RAMS, Moscow, Russia A.G. Groshev - Ph.D. (Biol.), Senior Research Scientist, Laboratory of Stromal Immu ne Regulation, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei of RAMS, Moscow, Russia I.L. Moskvina - Research Scientist, Laboratory of Stromal Immune Regulation, Institute of Epidemiology and Microbiology n.a. N.F. Gamalei of RAMS, Moscow, Russia

Cytokines are low molecular weight protein regulatory substances produced by a variety of cells such as endothelial cells, macrophages, neutrophils, lymphocytes, platelets, fibroblasts, et al., and able to modulate their functional activity. The stimulatory or inhibitory effects of cytokines made by bonding them with a large number of receptors on the cell surface and can be directed to the cells involved in inflammation, regeneration and development of the immune response. The work has studied cytokine ratio in 54 samples the conditioned medium obtained by culturing the multipotent mesenchymal stromal cells (MMSCI), human bone marrow, which allows to evaluate the production of cytokines by these cells. Isolation of MMSC and cultivation was performed according to known methods. The age of donors ranged from a few months to 70 years. In 5 cases, stromal cells were isolated of giant cell tumor (T-bills) patients. We investigated 7 samples of conditioned medium from the strains subjected to cryopreservation and subsequent development. In the above samples conditioned medium to study synthetic activity was obtained from cultures multiclonal strains. At the same time, undoubtedly of interest to submit data on the synthesis of cytokines monoclonal strains, the strains obtained as a result of proliferation of a stromal cell. We studied the profiles and levels of cytokine synthesis 6 monoclonal strains. Fence conditioned medium produced at different passages from 1 to 12 in terms of the development of strains of the 4 to 19 days. Determination of cytokines was carried out using the "Biopleks." We investigated the synthesis IL-1α, IL 4, 6, 8, 10, FGF, G-CSF, GM-CSF, MIP- 1b, MCAF, TNF-α, LIF, VEGF, MIF, SCF and others. A comparison of parametric indices performed using Student's t-test, the definition of communication parameters by using Pearson's correlation analysis. Results were considered accurate at p <0,05.ya. The cytokines was determined in pg/ml. Studies have shown that the synthesis of cytokines when cultured MMSK occurs with varying intensity. The most actively produced, such as the cytokines VEGF - 19881,5-49449,2 pg / ml, IL-6-1506,5-25815,2 pg/ml. Then, in order of decreasing concentration were IL-8, IL-10, MCAF, TNF-α, FGFb, GM-CSF, G-CSF, MIP-1b and has the lowest concentration of IL-4 (4,3-19,3) pg / ml. In first cultures of human bone marrow was investigated production of stem cell factor (SCF), FGFb- a major cytokines that enhance the proliferation of MMSK in vitro cytokine TNF-α. The same value product SCF (57,5-71,3 pg/ml) in primary cultures with the same time of culture, but a different number of explanted cells (3,0 × 105-2,0h106) may indicate a limitation of cytokine production by increasing the cell density . With further development of strains (1 to 8 passages) products of SCF increases from 104 to 258 pg/ml. Production of this cytokine strains MMSK after cryopreservation with the same number of explanted cells and the culture time (5 days) decreased and amounted to 50-90 pg / ml. Products FGFb decreases with age with 295-389 pg/ml in the first year to 97-111 pg/ml to 70 years, and does not depend on the density of explanted cells. Similar dynamics of production was typical for the cytokine TNF-α, decreasing from 1510-1561 pg / ml in the first year to 61-187 pg/ml to 70 years. Interesting results were obtained in studies of the synthesis of cytokines in cultures of monoclonal strains. Under the same conditions, the initiation and maintenance of crops (duration of cultivation, the number of explanted cells age donors) cytokine synthesis monoclonal strains did not differ from the synthetic activity of polyclonal strains. In comparing the average concentrations of the cytokines in groups of patients and healthy donors GKO was obtained a significant difference between them is only in the level MIP-1b, whose content in the conditioned medium of cell cultures GKO patients was 93.67 pg / ml compared to 25.53 pg / ml in healthy donors. Other indicators have not found significant differences. In the process of the research was found that MMSK synthetic activity declines with age. By synthetic activity were observed significant differences between the number of passages and the period of incubation of cell cultures. Explantation of increased amounts of MMSK in a limited amount of medium quickly deplete its reserves and reducing the synthetic potential, the more cells there are actively proliferate, which also requires a considerable amount of material for the synthesis.