Activation of regeneration of red and white pulp of the spleen after the combined transplantation of HSC and MSCS in terms of exposure to ionizing radiation

Author(s):  
И.Ю. Маклакова ◽  
Д.Ю. Гребнев ◽  
А.П. Ястребов

Цель - изучение влияния сочетанной трансплантации мультипотентных мезенхимальных стромальных (ММСК) и гемопоэтических стволовых клеток (ГСК), выделенных из плаценты, на регенерацию белой и красной пульпы селезенки в физиологических условиях и в условиях воздействия ионизующего излучения. Методика. Эксперименты выполнены белых лабораторных беспородных мышах-самцах. Облучение животных проводилось на гамма-терапевтической установке типа АГАТ-С с радионуклидным источником Co-60 типа ГИК-8-4, поглощенная доза составила 4,0 Гр, мощность поглощенной дозы 20 сГр/мин. Животным опытной группы внутривенно вводились аллогенные ММСК и ГСК соответственно в дозе 6 млн клеток/кг и 330 тыс. клеток/кг, суспендированные в 0,2 мл 0,9% раствора NaCl. Выделение гемопоэтических стволовых клеток осуществлялось методом прямой иммуномагнитной сепарации. Проводили морфометрию лимфоидных фолликулов селезенки (средняя площадь, средняя площадь В-зоны, средняя площадь герминативного центра, средняя площадь T-зоны), а также определялось среднее расстояние между центрами фолликулов и средняя клеточность красной пульпы. Результаты. Показано, что после воздействия ионизирующего излучения на фоне сочетанной трансплантации ММСК и ГСК происходит увеличение размеров лимфоидного фолликула за счет площади B-зоны фолликула, площади герминативного центра фолликула, восстановление содержания лимфобластов, пролимфоцитов и лимфоцитов до значений нормы. На фоне трансплантации ММСК и ГСК в условиях лучевой нагрузки установлено увеличение плотности клеток в красной пульпе селезенки и, как следствие, увеличение расстояния между центрами лимфоидных фолликулов. Увеличение плотности клеток в красной пульпе происходит как за счет увеличения содержания эритроидных клеток, так и за счет увеличения гранулоцитов. Заключение. Проведенные исследования свидетельствуют об эффективности сочетанной трансплантации ММСК и ГСК в отношении основных морфометрических показателей селезенки после воздействия ионизирующего излучения. The purpose of this work was to study the effect of combined transplantation of multipotent mesenchymal stromal (MSCS) and hematopoietic stem cells (HSCs) isolated from the placenta, on the regeneration of white and red pulp of the spleen under physiological conditions and in conditions of exposure to ionizing radiation. Methods. The experiments were performed with laboratory mice-males. We studied the influence of ionizing radiation dose of 4.0 Gy. Animals of the experimental group were intravenously infused into MMSC and GSK respectively at a dose of 6 million cells/kg and 330 thousand cells/kg, suspended in 0.2 ml of 0.9% NaCl solution. The selection of hematopoietic stem cells was carried out using the direct technique of immune magnetic separation. Were studied the following morphometric parameters of the spleen: the average area of lymphoid follicles, the average area of zone of lymphoid follicles, average size of germinal center of lymphoid follicles, average size T-zones of lymphoid follicles, the average distance between the centers of the follicles, the average cellularity of the red pulp. Results. As a result, of research obtained that after exposure to ionizing radiation on the background of combined transplantation of HSC and MSCS there is an increase in size of lymphoid follicle at the expense of area B-zone of the follicle, the area germinative center of the follicle, restoring the content of lymphoblasts and lymphoblasts and lymphocytes to normal values. On the background of transplantation MMSC and GSK in terms of radiation exposure changes and the red pulp of the spleen. The increase in the density of cells in the red pulp of the spleen and, as a consequence, of the increase of the distance between the centers of lymphoid follicles. The increase in the density of cells in the red pulp occurs due to the increase in the content of erythroid cells and by increasing granulocytes. Key words: ionizing radiation, multipotent mesenchymal stromal cells, hematopoietic stem cells, spleen, regeneration. Conclusion. Studies have shown the effectiveness of combined transplantation MSC and GSK in respect of the main morphometric parameters of the spleen after exposure to ionizing radiation.

2013 ◽  
Vol 94 (6) ◽  
pp. 911-914 ◽  
Author(s):  
D Yu Grebnev ◽  
A P Yastrebov ◽  
I Yu Maklakova

Aim. To study the changes of splenic morphometric parameters in aged laboratory animals exposed to ionizing radiation in the dose of 4Gr after multipotent mesenchymal and hematopoietic stem cells transplantation. Methods. The experiments were conducted on 72 white male laboratory mice at the third year of life with the body weight of 50 g. Multipotent mesenchymal stem cells and hematopoietic stem cells were obtained from 8 laboratory female mice with the body weight of 30 g aged 3-4 months, the gestation term was 14 days. The first group (36 animals exposed to radiation) was subdivided to 2 subgroups of 18 animals each. The suspension of multipotent mesenchymal stem cells (6 000 000 cells/kg) and hematopoietic stem cells (330 000 cells/kg) was introduced as single intravenous injection to the experimental subgroup (18 animals) 1 hour after the animals were exposed to radiation. The animals of control subgroup (18 animals) were injected 0.2 ml of normal saline. The second group included two subgroups 18 mice each that underwent the same procedure without being exposed to radiation. 9 animals from each group were withdrawn from the study at 1st and 7th day each. The lymphoid follicle gross area, area of the T-cell and B-cell zones, general numbers of cells in the red pulp of spleen, including erythrocyte and lymphocyte count, were measured in splenic histologic specimens using the morphometric «BioVision 2008» software. Results. It was shown that on the 7th day after exposure to ionizing radiation followed by stem cells transplantation, the area of thymus-independent zone of lymphoid follicle restored back to normal ranges. The effect of multipotent mesenchymal stem cells and hematopoietic stem cells transplantation also resulted in the increase of the number of cells in the red pulp of spleen. There were no significant changes observed in numbers of erythroid cells and white blood cells in the spleen red pulp compared to control subgroup. At the same time, the leukocyte number in the red pulp of spleen restored to normal values. Conclusion. The restoration of the basic morphometric parameters in spleen of aged laboratory animals exposed to ionizing radiation may be explained be increased homing of splenic colony-forming units with subsequent activation of extramedullary hematopoiesis in spleen, and apoptosis-reducing effect of multipotent mesenchymal stem cells.


Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2441-2441
Author(s):  
Diana Tronik-Le Roux ◽  
Johnny Nehme ◽  
Arthur Simonnet ◽  
Pierre Vaigot ◽  
Marie Anne Nicola ◽  
...  

Abstract Hematopoietic stem cells (HSC) are indispensable for the integrity of complex and long-lived organisms since they can reconstitute the hematopoietic system for life and achieve long term repopulation of lethally irradiated mice. Exposure of an organism to ionizing radiation (IR) causes dose dependant bone marrow suppression and challenge the replenishment capacity of HSC. Yet, the precise damages that are generated remain largely unexplored. To better understand these effects, phenotypic and functional changes in the stem/progenitor compartments of sublethally irradiated mice were monitored over a ten week period after radiation exposure. We report that shortly after sublethal IR-exposure, HSC, defined by their repopulating ability, still segregate in the Hoechst dye excluding side population (SP); yet, their Sca-1 (S) and c-Kit (K) expression levels are increased and severely reduced, respectively, with a concurrent increase in the proportion of SPSK cells positive for established indicators of HSC presence: CD150+/CD105+ and Tie2+. Virtually all HSCs quickly but transiently mobilize to replenish the bone marrow of myelo-ablated mice. Ten weeks after, whereas bone marrow cellularity has recovered and hematopoietic homeostasis is restored, major phenotypic modifications can be observed within the c-Kit+ Sca-1+ Lin−/low (KSL) stem/progenitor compartment: CD150+/Flk2− and Flk2+ KSL cell frequencies are increased and dramatically reduced, respectively. CD150+ KSL cells also show impaired reconstitution capacity, accrued γ-H2AX foci and increased tendency to apoptosis. This demonstrates that the KSL compartment is not properly restored 10 weeks after sublethal exposure, and that long-term IR-induced injury to the bone marrow proceeds, at least partially, through direct damage to the stem cell pool. Since thrombopoietin (TPO) has been shown to reduce haematopoietic injury when administered immediately after exposure to radiations, we asked whether TPO could restore the permanent IR-induced damage we observed in the HSC compartment. We first found in competitive transplant experiments that a single TPO administration rescued the impaired reconstitution capacity of HSC’s from animals exposed to sublethal IR. In addition, we observed that TPO injection right after irradiation considerably attenuates IR-induced long-term injury to the stem/progenitor compartment. Finally, the use of marrow cells from transgenic ubiquitous luciferase-expressing donors combined with bioluminescence imaging technology provided a valuable strategy that allowed visualizing HSC homing improvements of TPO-treated compared to untreated irradiated donors, and enabled the identification of a preferential cellular expansion sites which were inaccessible to investigation in most studies. Electronic microscopy analysis revealed that these sites show also differential activity of megakaryocytopoiesis with marked differences in the proplatelets reaching the vascular sinus. Altogether, our data provide novel insights in the cellular response of HSC to IR and the beneficial effects of TPO administration to these cells.


2019 ◽  
Vol 33 (7) ◽  
pp. 8138-8147 ◽  
Author(s):  
Hao Zeng ◽  
Mengjia Hu ◽  
Yukai Lu ◽  
Zihao Zhang ◽  
Yang Xu ◽  
...  

2011 ◽  
Vol 176 (6) ◽  
pp. 716-724 ◽  
Author(s):  
Satoru Monzen ◽  
Emiko Tashiro ◽  
Ikuo Kashiwakura

2020 ◽  
Vol 17 (2) ◽  
pp. 139-148
Author(s):  
I.Yu. Maklakova ◽  
◽  
V.V. Bazarniy ◽  
D.Yu. Grebnev ◽  
◽  
...  

The aim of this study was to study the changes in the morphological and functional state of the liver of old laboratory animals after its resection against the background of combined transplantation of multipotentmesenchymal stromal and hematopoietic stem cells.Materials and methods. The studies were performed on old laboratory animals, male mice of the age of 16-17 months. Resection of 2/3 of the liver was performed in old laboratory mice according to the method of C. Mitchell and H. Willenbring. One hour after subtotal resection, a combined transplantation of two types of cells was performed: multipotentmesenchymal stromal cells (at a dose of 4 million cells / kg) and hematopoietic stem cells (at a dose of 330 thousand cells / kg). The experiments on obtaining the MMSC and HSC culture were performed from the placenta chorion of 10 laboratory female mice 3-4 months old, weighing 20-25 g, gestational age 18 days. Assessment of liver regeneration was carried out on days 1, 3, 7 after resection of the liver by analyzing morphometric parameters of the liver and biochemical parameters of peripheral blood.Results. Studies have shown the activation of protein synthesizing liver function in old animals, a decrease in the level of cytolysis enzymes, an increase in the activity of cellular regeneration (increase in the hepatocyte nucleus area, the number of binuclear cells) of liver regeneration after resection against the introduction of stem cells.


2006 ◽  
Author(s):  
Hideyo Hirai ◽  
Pu Zhang ◽  
Tajhal Dayaram ◽  
Christopher Hetherington ◽  
Shin-ichi Mizuno ◽  
...  

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