EFFECT OF COMBINED TRANSPLANTATION OF MULTIPOTENT MESENCHYMAL STROMAL AND HEMATOPOIETIC STEM CELLS ON LIVER REGENERATION AFTER LIVER RESECTION IN OLD LABORATORY ANIMALS

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.

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.


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.


2020 ◽  
Vol 15 (3) ◽  
pp. 250-262
Author(s):  
Maryam Islami ◽  
Fatemeh Soleimanifar

Transplantation of hematopoietic stem cells (HSCs) derived from umbilical cord blood (UCB) has been taken into account as a therapeutic approach in patients with hematologic malignancies. Unfortunately, there are limitations concerning HSC transplantation (HSCT), including (a) low contents of UCB-HSCs in a single unit of UCB and (b) defects in UCB-HSC homing to their niche. Therefore, delays are observed in hematopoietic and immunologic recovery and homing. Among numerous strategies proposed, ex vivo expansion of UCB-HSCs to enhance UCB-HSC dose without any differentiation into mature cells is known as an efficient procedure that is able to alter clinical treatments through adjusting transplantation-related results and making them available. Accordingly, culture type, cytokine combinations, O2 level, co-culture with mesenchymal stromal cells (MSCs), as well as gene manipulation of UCB-HSCs can have effects on their expansion and growth. Besides, defects in homing can be resolved by exposing UCB-HSCs to compounds aimed at improving homing. Fucosylation of HSCs before expansion, CXCR4-SDF-1 axis partnership and homing gene involvement are among strategies that all depend on efficiency, reasonable costs, and confirmation of clinical trials. In general, the present study reviewed factors improving the expansion and homing of UCB-HSCs aimed at advancing hematopoietic recovery and expansion in clinical applications and future directions.


Author(s):  
Valentina Orticelli ◽  
Andrea Papait ◽  
Elsa Vertua ◽  
Patrizia Bonassi Signoroni ◽  
Pietro Romele ◽  
...  

2015 ◽  
Vol 39 (10) ◽  
pp. 1099-1110 ◽  
Author(s):  
Iordanis Pelagiadis ◽  
Eftichia Stiakaki ◽  
Christianna Choulaki ◽  
Maria Kalmanti ◽  
Helen Dimitriou

Blood ◽  
1997 ◽  
Vol 89 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Futoshi Hashimoto ◽  
Kikuya Sugiura ◽  
Kyoichi Inoue ◽  
Susumu Ikehara

Graft failure is a mortal complication in allogeneic bone marrow transplantation (BMT); T cells and natural killer cells are responsible for graft rejection. However, we have recently demonstrated that the recruitment of donor-derived stromal cells prevents graft failure in allogeneic BMT. This finding prompted us to examine whether a major histocompatibility complex (MHC) restriction exists between hematopoietic stem cells (HSCs) and stromal cells. We transplanted bone marrow cells (BMCs) and bones obtained from various mouse strains and analyzed the cells that accumulated in the engrafted bones. Statistically significant cell accumulation was found in the engrafted bone, which had the same H-2 phenotype as that of the BMCs, whereas only few cells were detected in the engrafted bones of the third-party H-2 phenotypes during the 4 to 6 weeks after BMT. Moreover, the BMCs obtained from the MHC-compatible bone showed significant numbers of both colony-forming units in culture (CFU-C) and spleen colony-forming units (CFU-S). These findings strongly suggest that an MHC restriction exists between HSCs and stromal cells.


Stem Cells ◽  
2001 ◽  
Vol 19 (1) ◽  
pp. 46-58 ◽  
Author(s):  
Kikuya Sugiura ◽  
Hiroko Hisha ◽  
Junji Ishikawa ◽  
Yasushi Adachi ◽  
Shigeru Taketani ◽  
...  

Blood ◽  
1997 ◽  
Vol 89 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Futoshi Hashimoto ◽  
Kikuya Sugiura ◽  
Kyoichi Inoue ◽  
Susumu Ikehara

Abstract Graft failure is a mortal complication in allogeneic bone marrow transplantation (BMT); T cells and natural killer cells are responsible for graft rejection. However, we have recently demonstrated that the recruitment of donor-derived stromal cells prevents graft failure in allogeneic BMT. This finding prompted us to examine whether a major histocompatibility complex (MHC) restriction exists between hematopoietic stem cells (HSCs) and stromal cells. We transplanted bone marrow cells (BMCs) and bones obtained from various mouse strains and analyzed the cells that accumulated in the engrafted bones. Statistically significant cell accumulation was found in the engrafted bone, which had the same H-2 phenotype as that of the BMCs, whereas only few cells were detected in the engrafted bones of the third-party H-2 phenotypes during the 4 to 6 weeks after BMT. Moreover, the BMCs obtained from the MHC-compatible bone showed significant numbers of both colony-forming units in culture (CFU-C) and spleen colony-forming units (CFU-S). These findings strongly suggest that an MHC restriction exists between HSCs and stromal cells.


Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1392-1392
Author(s):  
Yoko Okitsu ◽  
Hideo Harigae ◽  
Masanori Seki ◽  
Toru Fujiwara ◽  
Shinichiro Takahashi ◽  
...  

Abstract (Introduction) Aplastic anemia (AA) is characterized by peripheral pancytopenia and fatty bone marrow. An immunological attack to hematopoietic stem cells has been thought to be responsible for the development of the disease. Previously, we reported the expression of transcription factor GATA-2 is significantly decreased in CD34 positive cells in AA. Together with the phenotypes of hematopoietic stem cells in GATA-2 hetero-knockout mice, GATA-2 down-regulation may play a role in the reduction of a stem cell pool observed in AA. On the other hand, GATA-2 has been shown to be essential for the maintenance of immaturity of preadipocytes. If a pathological immune response in AA decreases the level of GATA-2 expression in not only hematopoietic stem cells but also stromal preadipocytes, it may accelerate the maturation of preadipocytes, leading to the formation of fatty bone marrow. To explore this possibility, the phenotypic change of stromal preadipocytes by suppression of GATA-2 was examined in this study. (Method) The GATA-2 expression level was suppressed by using siRNA for GATA-2 in mouse stromal preadipocyte cell lines, TBR9 and TBR343. After the treatment with siRNA, the adipocyte differentiation was induced by the incubation with insulin and dexamethasone for 7days. Then, the maturation level was examined by oil drops formation judged by oil red staining, and by the expression level of adipcin and PPAR-γ mRNA. Supporting activity of hematopoietic colony formation was also evaluated by using mouse fetal liver cells after siRNA treatment. (Results) By using designed siRNA, the GATA-2 expression was suppressed to 30% of control, whereas the expression level of GATA-3, which is co-expressed in preadipocytes, was unchanged. When GATA-2 was suppressed by siRNA, the oil drop formation and adipocyte-specific gene expression was significantly accelerated in both of stromal cells. Furthermore, the number of fetal liver hematopoetic colonies was significantly decreased by suppression of GATA-2, suggesting that GATA-2 down-regulation in stromal preadipocytes results in not only the acceleration of the maturation but also the reduced supporting activity of hematopoietic colony formation (Conclusion) These results suggest that suppression of GATA-2 in hematopoietic tissues induces the characteristic features of AA, i.e., decreased the number of hematopoietic stem cells and increased number of mature adipocytes.


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