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The aim of the work was to study the impact of fetal rat brain cell supernatant (FRBCS) on the expression of transforming growth factor β1 (TGF-β1) and p53 in C6 cells of rat glioma in vitro. Materials and Methods: FRBCS was obtained from suspensions of fetal rat brain cells on the 14th (E14) day of gestation. C6 glioma cells were cultured for 48 h in the presence of FRBCS or FRBCS + anti-TGF-β1 monoclonal antibody. Immunocytochemical staining for TGF-β1 and p53 was performed. Results: The proportion of TGF-β1-immunopositive tumor cells in C6 glioma cultures was statistically significantly higher than in the control cell cultures of normal fetal rat brain. FRBCS reduced the proportion of TGF-β1-immunopositive tumor cells and increased the proportion of p53-immunopositive cells in C6 glioma cultures. In cells cultured with FRBCS + anti-TGF-β1 monoclonal antibody, the above effects of FRBCS were abrogated. Conclusion: The obtained results suggest that TGF-β1 seems to be responsible for decrease in TGF-β1 expression and increase in p53 expression in C6 glioma cells treated with FRBCS.

Platelets play an important part in the progression and pathological angiogenesis of brain glioma because of the different granules content and release of microvesicles that are the source of numerous mediators and bioactive substances, which probably provides a "strategy" for the tumour survival. The objective of study was exploring the effect of platelet-released secretion products of patients with brain glioma on the experimental model of tumour growth in vitro. For this purpose, the cells of glioma C6 were cultured for 72 hours under the addition of modified media containing platelet-released secretion products or conditioned media of peripheral blood cells of patients with glioma as well as persons of the comparison group without rough somatic pathology. In control glioma C6 cultures in standard conditions cell clusters were formed by the type of "spheroids", from which radial cell migration occurred, a tense cellular or reticular growth zone was formed, and tumour cells preserved their ability to mitotic division. Under the influence of platelet-released secretion products of patients with glioma, differently directed effects on cell mitotic activity and the number of cell clusters in glioma C6 cultures were detected depending on the degree of tumour malignancy: stimulating effect under the influence of platelet factors of patients with high-malignancy glioma (G4) and inhibitory effect – due to the influence of platelet factors of patients with differentiated glioma (G2). In contrast to the thrombocyte-released factors, the conditioned media of a common pool of peripheral blood cells of patients with G4 glioma suppressed the mitotic activity of tumour cells and did not affect the number of cell clusters. No changes in glioma C6 cultures were revealed after the influence of platelet-released secretion products of persons of the comparison group. The obtained data confirm the important role of platelets in the pathogenesis of brain glioma, pointing to the fundamental difference in the spectrum of biologically active molecules that are released by platelets of patients depending on the degree of tumour malignancy and are able to regulate the cell cycle and proliferative activity of the glioma tumour cells, which may have application as a diagnostic marker as well as predictive marker of response to antitumour therapy.

The transplantation of multipotent mesenchymal stromal cells (MMSCs) is considered to be a possible therapy of multiple sclerosis. For the clinical application of human umbilical cord-derived MMSCs (UC-MMSCs) it is necessary to develop a method of their cryopreservation taking into account the type of cryoprotective media and to investigate the possibility of using these cells for therapeutic purposes in vivo. The purpose of the study was to investigate the effect of UC-MMSCs, cryopreserved in solutions of different composition, on the processes of demyelination and remyelination of the spinal cord of rats with experimental allergic encephalomyelitis (EAE) as a model of multiple sclerosis. Materials and methods. The EAE was modeled by subcutaneous administration of homogenized spinal cord of adult rats with complete Freund's adjuvant. On the 18th day rats with moderate relapsing-remitting form of EAE were suboccipitally injected 1•106 UC-MMSCs, cryopreserved in cryoprotective media containing dimethyl sulfoxide (DMSO), fetal bovine serum (FBS), ethylene glycol, trehalose and sucrose at different composition. On the 35th and 60th days, the studies of ultrastructural changes of the lumbar spinal cord (L3-L5) were performed, assessing the degree of demyelination of nerve fibers by the ratio of myelin sheath (MS) thickness to the diameter of the axis cylinder (AC) of axons. Results. In rats with moderate EAE from the 35th to the 60th day after the modelling of the disorder, destructive changes and signs of demyelination in the spinal cord increased; the MS/AC index corresponded to the average degree of axon demyelination. Suboccipitally administered cryopreserved UC-MMSCs to EAE rats, depending on the used cryopreservation solution, slowed or stopped the demyelination, decreased the MS/AC index to a low degree of axonal demyelination. Reducing the concentration of DMSO in the cryopreservation medium from 10 % to 4 % and adding 6 % trehalose provided a better effectiveness of UC-MMSCs in decreasing the degree of demyelination in EAE. At the same time, the standard solution (10 % DMSO, 90 % FBS) provided these effect, but to a lesser extent. The use of a multicomponent cryopreservation medium containing 15 % ethylene glycol, 3 % DMSO, 10 % sucrose, 12 % trehalose and 60 % FBS did not achieve the goal of maintaining the effects of UC-MMSCs to reduce the degree of demyelination in EAE. Conclusions. To maintain the therapeutic properties of UC-MMSCs, it is advisable to add a reduced concentration of DMSO (4 %) and 6 % trehalose to the cryopreservation medium, supplemented with 90 % fetal bovine serum.