Comparison the impact of mesenchymal stromal cells, their microvesicles and plasma enriched with soluble platelet factors on survival and apoptosis of rat spleen lymphocytes in vitro Oksana V. Klimenkova, Michael P. Potapnev, Natalia V. Goncharova, Oleg A. Kudelich, Gennady G. Kondratenko Genes and Cells, 2024 BACKGROUND: The trophic function is one of the important but insufficiently studied features of mesenchymal stromal cells (MSCs), their microvesicles (MVs), and plasma enriched with soluble platelet factors (PRPs), which ensure the viability of target cells.AIM: To analyze the capability of MSCs, MVs, and PRP to affect the viability and spontaneous and activation-induced apoptosis of rat spleen lymphocytes during culturing in vitro.MATERIALS AND METHODS: MSCs were isolated from the mononuclear cell fraction of the femoral bone marrow of Wistar rats using the plastic adhesion method. MVs were obtained from the conditioned medium of MSCs by centrifugation at 14,500 g. PRP was prepared by freezing/thawing rat peripheral blood platelet concentrate. Rat spleen lymphocytes were isolated on a density gradient of 1.077 g/cm3. The viability and degree of lymphocyte apoptosis in vitro were assessed by flow cytometry for 7-AAD incorporation and binding to annexin V.RESULTS: The presence of MSCs at 10 and 20% concentrations caused an increase in the number of living intact and PMA-activated lymphocytes by the end of 3-day in vitro cultivation. Compared with controls, the amount of necrotic cells decreased 8.3–13.5 times, and the number of apoptotic cells decreased 2.3–4.0 times, mainly due to lymphocytes at the late stage of apoptosis. MVs of MSCs at the indicated concentrations did not significantly affect the viability of lymphocytes cultured in vitro but reduced the level of apoptosis of intact and PMA-activated lymphocytes by 3.6 (р=0.03) and 4.8–5.2 (р=0.048; р=0.03) times respectively. Studies have shown that 1.25% rat PRP has a growth-stimulatory activity against MSCs but not against lymphocytes cultured in vitro. In the culture of intact lymphocytes, PRPs did not significantly affect cell viability with a slight decrease (2.7–2.9 fold, p 0.05) in the number of necrotic cells. In the culture of PMA-activated lymphocytes, 1.25–2.50% PRP increased the number of living cells by 1.6–2.2 times (р=0.002; р=0.01) and the number of necrotic cells by two times (р=0.02).CONCLUSION: MSCs, MVs of MSCs, and PRP, in decreasing order, enhanced the viability of rat spleen lymphocytes and suppressed late apoptosis during in vitro cultivation. Lymphocytes activated by phorbol myristate acetate are more sensitive to their vital action compared with resting cells.
Regional use of extracellular microvesicles of mesenchymal stromal cells in acute necrotizing pancreatitis in an experiment O. A. Kudelich, G. G. Kondratenko, M. P. Potapnev, O. V. Klimenkova, N. V. Goncharova Transplantologiya the Russian Journal of Transplantation, 2024 Background. The significance of the problem of treatment of acute pancreatitis is due to an increase in the incidence with an increase in the number of necrotizing forms, accompanied by a high incidence of severe complications and high mortality.Objective. To identify the impact of regionally used extracellular microvesicles of mesenchymal stromal cells on the endogenous intoxication markers in acute necrotizing pancreatitis in the experiment.Material and methods. Acute pancreatitis was induced by the introduction of a 0.3 ml of 5% solution of non-ionic polyethylene glycol octylphenol ether detergent into the caudal part of the rat pancreas. The study was conducted on 42 adult Wistar rats, which were randomly divided into 4 groups. Group I (n=6) included intact animals, Group II (control group) (n=12) included rats with pancreatitis without treatment, Group III (n=12) consisted of rats with pancreatitis treated with analgesia + infusions of 0.9% sodium chloride solution (saline), Group IV (n=12) included rats with pancreatitis treated with analgesia+ saline infusions + regional application of extracellular microvesicles of mesenchymal stromal cells. Cells were obtained from the bone marrow of healthy animals. Microvesicles were obtained by differential centrifugation under sterile conditions. Microvesicles were administered one day after the pancreatitis induction through the catheter installed into the pathologically altered part of the pancreas. The dose of microvesicles was calculated as equivalent to (derived from) 1 million mesenchymal stromal cells. The hematological parameters, markers of the systemic manifestation of the pathological process (alpha-amylase, aspartate aminotransferase, alanine aminotransferase), the endogenous intoxication markers (lipid peroxidation activity, nitric oxide level), the systemic inflammatory response markers (tumor necrosis factor-alpha, interleukin-6) were studied on the 3rd and 7th day from the start of disease modeling.Results. Regional use of extracellular microvesicles of mesenchymal stromal cells in the treatment of acute experimental necrotizing pancreatitis at an early stage helped to normalize the level of blood platelets, reduce enzymeemia, elements of endogenous intoxication (interleukin-6, tumor necrosis factor-alpha), and the nitric oxide level.Conclusion. The early application of extracellular microvesicles of mesenchymal stromal cells in the treatment of acute necrotizing pancreatitis in an experiment has a positive effect on parameters, which are key links of pathogenesis and leading markers of this disease severity.
Comparative Evaluation of Cellular Origin Bioproducts Effects on the Course of Acute Necrotizing Pancreatitis in Experiment О. А. Куделич, Г. Г. Кондратенко, Михаил Петрович Потапнев, Татьяна Сергеевна Колесникова, О. В. Клименкова, et al. Surgery Eastern Europe, 2024 Цель. Обосновать возможность коррекции системной патологической реакции при экспериментальном остром некротизирующем панкреатите с помощью локального применения мезенхимальных стромальных клеток (МСК), их микровезикул (МВ) или плазмы, обогащенной растворимыми факторами тромбоцитов (ПОРФТ). Материалы и методы. Изучено влияние МСК, МВ МСК и ПОРФТ на гематологические и биохимические показатели при остром экспериментальном некротизирующем панкреатите. Исследование проведено на 54 половозрелых крысах-самцах линии Wistar. Модель острого некротизирующего панкреатита создавали стандартно введением 0,3 мл 5% неионного детергента полиэтиленгликоль-октилфенолового эфира в хвостовую часть поджелудочной железы крыс. На 3-и и 7-е сутки от начала моделирования заболевания оценивали гематологические показатели, маркеры системного проявления патологического процесса (альфа-амилаза, глюкоза, аспартатаминотрансфераза, аланинаминотрансфераза), маркеры эндогенной интоксикации (малоновый диальдегид, уровень оксида азота), маркеры системного воспалительного ответа (фактор некроза опухоли α, интерлейкин-6, СРБ). Результаты. Применение МСК и их микровезикул на ранней стадии острого некротизирующего панкреатита положительно влияет на количество тромбоцитов в периферической крови экспериментальных животных. В эти же сроки МСК предотвращали гипергликемию, что может свидетельствовать о цитопрототекторном их действии в отношении гормонпродуцирующих клеток поджелудочной железы. Использование МСК и ПОРФТ способствует нормализации активности α-амилазы в сыворотке крови, что также может косвенно свидетельствовать о системном антиферментном, опосредованном через цитопротекторное действие в отношении экзокринных клеточных структур поджелудочной железы. Использование ПОРФТ, МСК и МВ МСК при остром экспериментальном некротизирующем панкреатите уменьшает системный воспалительный ответ, что продемонстрировала динамика значений ФНО-α и ИЛ-6. В сыворотке крови животных, которым проводилось лечение МСК, уровень СРБ на протяжении всего эксперимента не повышался. Регионарное использование МСК препятствует росту уровня оксида азота и малонового диальдегида в сыворотке крови экспериментальных животных. Заключение. Раннее регионарное применение биопродуктов клеточного происхождения при экспериментальном остром некротизирующем панкреатите оказывает системный положительный эффект, что позволяет рассматривать их в качестве перспективных средств для патогенетического лечения данного заболевания. Purpose. To substantiate a possibility of correcting a systemic pathological reaction in experimental acute necrotizing pancreatitis by local application of mesenchymal stromal cells (MSCs), their microvesicles (MV MSCs), or plasma enriched with platelet factors (PRP). Materials and methods. The effect of MSCs, MB MSCs and PORFT on hematological and biochemical parameters in acute experimental necrotizing pancreatitis was studied. The study was carried out on 54 adults Wistar rats. Acute pancreatitis was induced by introducing 0.3 ml of 5% solution of non-ionic polyethylene glycol octylphenol ether detergent into the caudal part of rats’ pancreas. Hematological parameters, markers of systemic manifestation of the pathological process (alpha-amylase, glucose, AST, ALT), markers of endogenous intoxication (malondialdehyde, nitric oxide), and markers of systemic inflammatory response (tumor necrosis factor-alpha, interleukin-6, CRP) were evaluated on the 3rd and 7th day from the beginning of disease modeling. Results. MSCs and their microvesicles used at early stages of acute necrotizing positively affect the number of platelets in the peripheral blood of experimental animals. At the same time, MSCs prevented hyperglycemia, indicating their cytoprotective effect on hormone-producing pancreatic cells. The use of MSCs and PRP promotes normalization of α-amylase activity in serum, which may also indirectly indicate systemic anti-enzyme action, mediated through cytoprotective one, against exocrine cellular structures of the pancreas. MSCs, MB MSCs and PRP application in acute experimental necrotizing pancreatitis reduced the systemic inflammatory response, as demonstrated by TNF-α and IL-6 values changes. CRP blood serum levels in animals treated with MSCs did not increase throughout the experiment. A regional use of MSCs prevents increasing nitric oxide and malondialdehyde levels in blood serum of experimental animals. Conclusion. A regional use of cell origin bioproducts at early stages of experimental acute necrotizing pancreatitis provides a positive systemic effect, which allows considering them as promising agents for pathogenetic treatment of this disease.
In vitro expansion and lineage commitment of the human umbilical cord blood myeloid progenitors Cellular Transplantation and Tissue Engineering, 2012
Non-viral engineering of skin precursor-derived Schwann cells for enhanced NT-3 production in adherent and microcarrier culture A. Shakhbazau, D. Shcharbin, M. Bryszewska, R. Kumar, H. M. Wobma, et al. Current Medicinal Chemistry, 2012 Genetic engineering of stem cells and their derivatives has the potential to enhance their regenerative capabilities. Here, dendrimer- and lipofection-based approaches were used for non-viral neurotrophin-3 (NT-3) over-expression in Schwann cells differentiated from skin precursors (SKP-SCs). A variety of dendrimers were first tested for transfection efficiency on HEK 293T cells, with PAMAMNH2 G4 found most effective and used subsequently for SKP-SCs transfection. Plasmid-based expression resulted in increased NT-3 release from SKP-SCs in both adherent and microcarrier-based culture. In a proof-of-concept study, the microcarrier/SKP-SCs were implanted into the injured nerve, and transfected cells were shown to detach, integrate into the nerve tissue and associate with regenerating axons. Virus-free systems for transient neurotrophin expression are a feasible and biologically safe option to increase the therapeutic value of stem cells and stem cell-derived cells in nerve repair strategies. Further work to develop bioprocesses for expansion of SKP-SCs on microcarriers in bioreactors is still needed.
Modeling long-haul WDM transmission in nonlinear optical fibers using a fast split-step wavelet collocation method Piers 2004 Progress in Electromagnetics Research Symposium Extended Papers Proceedings, 2004
Comparison the impact of mesenchymal stromal cells, their microvesicles and plasma enriched with soluble platelet factors on survival and apoptosis of rat spleen lymphocytes in … OV Klimenkova, MP Potapnev, NV Goncharova, OA Kudelich, ... Genes & Cells 19 (3), 387-399 , 2024 2024
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MARKERS OF PLATELET DYSFUNCTION IN THE FIRST 48 HOURS OF LACUNAR STROKE RESULTING FROM CEREBRAL SMALL VESSEL DISEASE L Anatskaia, N Goncharova, L Matusevich, N Shcharbina, V Zabarovski, ... INTERNATIONAL JOURNAL OF STROKE 16 (2_ SUPPL), 50-50 , 2021 2021
Clinical benefits of single vs repeated courses of mesenchymal stem cell therapy in epilepsy patients F Hlebokazov, T Dakukina, M Potapnev, S Kosmacheva, L Moroz, ... Clinical neurology and neurosurgery 207, 106736 , 2021 2021 Citations: 37
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The features of cellular immunity in patients with treatment-resistant asthma Z Antanovich, N Goncharova European Respiratory Journal 50 (suppl 61) , 2017 2017
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P-GLYCOPROTEIN TRANSPORT ACTIVITY UNDER REDOX BALANCE CHANGES IN LYMPHOCYTES OF PATIENTS WITH B-CHRONIC LYMPHOCYTIC LEUKEMIA Y Harmaza, A Tamashevski, N Goncharova, E Slobozhanina, ... HAEMATOLOGICA 101, 721-721 , 2016 2016
Our experience in application of autologous bone-marrow-derived mesenchymal stem cells for therapy of patients with symptomatic epilepsy FP Hlebokazov, TV Dоkukina, SI Ighnatenko, SM Кosmacheva, ... Epilepsy and paroxysmal conditions 6 (1), 6-14 , 2016 2016 Citations: 2
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Clinical phenotype of destructive-regenerative endothelial response in the first 48 hours of lacunar stroke resulting from cerebral small vessels disease L Anatskain, N Goncharova, G Khulup CEREBROVASCULAR DISEASES 41, 198-198 , 2016 2016
EFFICIENCY OF MONTELUKAST TREATMENT OF THE CHILDREN, SUFFERING FROM BRONCHIAL ASTHMA II Balabolkin, IE Smirnov, AV Liapunov, OF Lukina, NV Goncharova, ... Current Pediatrics 5 (5), 35-38 , 2015 2015
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Effect of intravenous low intensity laser irradiation of blood on the level of angiogenic T-cells in acute lacunar stroke L Anatskaia, N Goncharova, I Seviaryn, GG Khulup, SS Marchenko, ... EUROPEAN JOURNAL OF NEUROLOGY 22, 808-808 , 2015 2015
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Influence of intravenous low-intensity laser irradiation of blood on the level of circulating endothelial progenitor cells in acute lacunar stroke L Anatskaia, N Goncharova, I Seviaryn, G Khulup INTERNATIONAL JOURNAL OF STROKE 10, 367-367 , 2015 2015
MOST CITED SCHOLAR PUBLICATIONS
Treatment of refractory epilepsy patients with autologous mesenchymal stem cells reduces seizure frequency: an open label study F Hlebokazov, T Dakukina, S Ihnatsenko, S Kosmacheva, M Potapnev, ... Advances in medical sciences 62 (2), 273-279 , 2017 2017 Citations: 69
Investigation of morphometric parameters<? xpp qa?> for granulocytes and lymphocytes as applied<? xpp qa?> to a solution of direct and inverse light-scattering problems GI Ruban, SM Kosmacheva, NV Goncharova, DV Bockstaele, VA Loiko Journal of biomedical optics 12 (4), 044017-044017-11 , 2007 2007 Citations: 61
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Morphometric model of lymphocyte as applied to scanning flow cytometry VA Loiko, GI Ruban, OA Gritsai, AD Gruzdev, SM Kosmacheva, ... Journal of Quantitative Spectroscopy and Radiative Transfer 102 (1), 73-84 , 2006 2006 Citations: 48
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Experimental search for new means of Pathogenetic therapy COVID-19: Inhibitor of H2-receptors famotidine increases the effect of oseltamivir on survival and immune status of … NV Goncharov, KA Vasilyev, IV Kudryavtsev, PP Avdonin, DA Belinskaia, ... Journal of Evolutionary Biochemistry and Physiology 58 (1), 230-246 , 2022 2022 Citations: 2
Our experience in application of autologous bone-marrow-derived mesenchymal stem cells for therapy of patients with symptomatic epilepsy FP Hlebokazov, TV Dоkukina, SI Ighnatenko, SM Кosmacheva, ... Epilepsy and paroxysmal conditions 6 (1), 6-14 , 2016 2016 Citations: 2
Growth-stimulating activity of platelet preparations in relation to mesenchymal stem cells in vitro SI Ihnatsenko, SM Kasmachova, MP Potapnev, EA Kahno, ... Proceedings of the National Academy of Sciences of Belarus, Medical series … , 2016 2016 Citations: 2
Investigation of human white blood cells: application to scanning flow cytometry VA Loiko, GI Ruban, OA Gritsai, AA Miskevich, SM Kosmacheva, ... International Conference on Lasers, Applications, and Technologies 2005 … , 2006 2006 Citations: 2
Discrepancy between direct and antibody-dependent cytotoxic activities of human LAK cells MP Potapnev, TS Garbuzenco, NV Goncharova, VD Zobnin, OV Shadrin, ... Immunology letters 41 (1), 13-17 , 1994 1994 Citations: 2
Dual Effect of Amino Acid Compositions on Antibacterial Activity of Human Neutrophilic Granulocytes MP Potapnev, SV Andreyev, NV Goncharova, OI Viatkina, EL Berdina, ... Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry 17 (1), 17-25 , 2023 2023 Citations: 1
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