Перегляд за Автор "Kladnytska L.V."
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Документ Functional activity and morphological peculiarities of mesenchimal stem cells during in vitro cultivation conditions(ВЦ НУБіП України, 2018) Kladnytska L.V.; Mazurkevych A.I.; Khomych V.T.; Mazurkevych T.A.; Stegney Z.G.; Maluk М.О.; Garmanchuk L.V.; Velychko S.V.; Kharkevych Y.O.; Shelest D.V.; Velychko V.S.; Stupak В.The studies were conducted on 2-3-months-old males of C57BL/6 mice weighing 20-24 g. Obtaining and cultivating of mesenchimal stem cells (MSCs) were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. MSCs of the 2, 4, 7 and 12 passages were analyzed. Morphometric analysis was performed using a light microscopy. Morphometric parameters such as cell and nucleus area or nuclear-cytoplasmic ratio (NCR) were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and ImageJ 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological features of cells during cultivation changes: at first cells have a spindle-like shape with two long cytoplasmic processes, located bipolar. In later passages, cells have a significant number of cytoplasm processes, bipolar arrangement of processes changes to stellar. The NCR index of MSC significant decreases at the 4 passage by 12,9 % (p ≤ 0,05), at the 7 passage - by 35,3 % (p <0,001), at the 12 passage - by 76,6 % (p <0,001) compared to the initial state. The proliferative activity of the MSC of the bone marrow during cultivation significantly dereases at the later passages. Cell resistance to apoptosis induced by cultivation in the serum-free medium is fairly high. The number of cells in the state of apoptosis was 14,0±1,74 at the 4 passage and was reliably increased at the 12 passage to 22,67±1,55 % (p ≤ 0,05) during cultivation.Документ The influence of allogeneic bone marrow derived mesenchymal stem cells on indicators of functional state of immune organs in mice C57BL/6(ВЦ НУБіП України, 2018) Kladnytska L.V.; Mazurkevych A.I.; Velychko S.V.; Kharkevych Y.O.; Shelest D.; Velychko V.S.The studies were conducted on 2-3-months-old males of mice weighing 20-24 g. Оur work was to study the functional state of the organs of the immune system of C57Bl/6 mice after introduction of allogeneic MSCs of bone marrow origin. Obtaining and cultivating of MSCs were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. C57Bl/6 mice bone marrow aspirate cultured in a CO2 incubator at 37 оC and 5 % CO2 in DMEM with 10-15 % of fetal bovine serum, 1 % of antibiotic-antimycotic solution (Sigma-Aldrich, USA). The following groups of animals were formed: 1 group – intact (control group); 2 group - animals, to whom 0.5 ml of 0.9 % NaCl solution (placebo) were injected into the caudal vein; 3 group – animals, to whom 104 of allogeneic MSCs in 0.5 ml of phosphate buffer solution were injected into the caudal vein. The weight index, cellularity of thymus and spleen in C57Bl/6 mice investigated after the introduction of MSCs. The administration of allogenic mesenchymal stem cells of the bone marrow origin affects on the central and peripheral organs of the immune system. Administration of allogenic mesenchymal stem cells cause a significant increase in the content of lymphoid cells in the thymus at 7 and 18 days by 72 and 39 %, respectively (p < 0.01, p < 0.05) compared to the control. Administration of allogenic mesenchymal stem cells cause a significant increase in the weight index of the spleen and its cellularity at the 7 and 18 days of the immune response by 26 and 17 %, respectively (p < 0.01, p <0.05) compared to the control. the weight index of the spleen and it cellularity a significant increase at the 7 and 18 days of the immune response by 26 and 17 %, respectively (p < 0.05) compared to the control after administration of allogenic mesenchymal stem cells.