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The multipotent stromal cells (MSCs) are considered as one of the most promising agents for regenerating the immune system due to its powerful secretion of reparative factors and immunomodulatory properties. The PURPOSE of the study is to investigate the effect of co-transplantation of bone marrow hematopoietic stem cells (HSCs) and thymic multipotent stromal cells (MSCs) on regeneration of murine immune system damaged by cyclophosphamide. MATERIALS AND METHODS. MSCs were obtained from thymuses of C57BL mice using explant technique. Bone marrow cells (BMCs) were obtained by flushing out the femur with nutrient medium. The immune deficiency of mice was modelled by the treatment with cyclophosphamide. After that, the cells were co-transplanted and the parameters of the immune system were evaluated. We determined the total number of erythrocytes, hematocrit, and hemoglobin concentration in peripheral blood; phases of the cell cycle and apoptosis of the cells of the mesenteric lymph nodes; the number of antibody-producing cells in the spleen; delayed type hypersensitivity (DTH); proliferative and cytotoxic activity of natural killer lymphocytes; phagocytic activity, level of spontaneous and induced bactericidal activity of peritoneal macrophages. RESULTS. It was shown that in contrast to BMCs, the use of MSCs alone or co-transplantation of these cells increased the spontaneous proliferative activity of lymphocytes with a significant decrease in the number of lymph node cells in G0/G1 phase by 9.2 % and an increase in the number of lymphocytes in G2-M+S phase by 35 %, as well as restoring cellularity of bone marrow, thymus and lymph nodes in mice treated with cyclophosphamide. Regeneration of erythropoiesis was stimulated by BMCs, which was manifested by the normalization of hematocrit and hemoglobin, and an increase in the number of reticulocytes in the blood by 2.2 times compared with the group of mice receiving cyclophosphamide. Co-transplantation had less pronounced, but similar effect. Transplantation of thymic MSCs stimulated the natural cytotoxicity of splenocytes by 2.7 times and substantially increased the number of antibody-producing cells in the spleen by 1.7 times compared with the group of mice receiving cyclophosphamide. Co-transplantation had a more pronounced suppressive effect on the blast transformation reaction induced by phytohemagglutinin by 1.7 times, but showed a stimulating effect on DTH response by 1.46 times. Transplantation of BMCs did not affect the functional activity of the immune system. CONCLUSION. The effects of co-transplantation of BMCs and thymic MSCs are realized in the several parts: stimulation of hematological parameters recovery (like under the effect of BMCs separately), normalization of cell number of lymphoid organs (as under the impact of thymic MSCs); inhibition of blast transformation activity and stimulation of DTH are the effects of co-transplantation.

klìtinna ta organna transplantologìâ

The effects of co-transplantation of bone marrow hematopoietic stem cell and thymic multipotent stromal cells on the immune system of mice during its regeneration after cyclophosphamide treatment