Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells
Fumio NakaharaDaniel K. BorgerQiaozhi WeiSandra PinhoMaria MaryanovichAli H. ZahalkaMasako SuzukiCristian D. CruzZichen WangChunliang XuPhilip E. BoulaisAvi Ma’ayanJohn M. GreallyPaul S. Frenette
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Objective To study the difference of mRNA expression level of hematopoietic growth factors between human umbilical cord stromal cells and bone marrow stromal cells in vitro.Methods We detected the mRNA expression level of GM-CSF,SCF,TPO of human umbilical cord blood and bone marrow stromal cells being cultured for 28d in vitro.Results Human umbilical cord blood stromal cells secrete more TPO and less SCF,GM-SCF than bone marrow stromal cells.Conclusion Human umbilical cord blood stromal cells and bone marrow stromal cells could expressi mRNA of hematopoietic growth factors such as GM-CSF,SCF,TPO.They all may regulate hematopoiesis through secreting multiple hematopoietic growth factors.
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The role of thymus in the regulation of stromal elements, responsible for haemopoiesis inducing microsurrounding transfer of animals, subjected to 10-hours immobilization, was studied. The development of bone marrow hyperplasia and stimulation of functional activity of stromal cells, responsible for haemopoiesis inducing microsurrounding transfer, were shown to be thymus dependent processes during stress.
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Radiosensitivity of hemopoietic stroma precursors from a long-term culture of murine bone marrow, as measured by the adherent cell layer implantation techniques, was characterized by D0 = 3.02 +/- 0.7 Gy and n = 1.6. Mature cells of the hemopoietic microenvironment survived after doses of up to 100 Gy. Their irreversible damage was only observed after 150-200 Gy irradiation. The results obtained support the suggestion of different histogenetical origin of the hemopoietic and stromal precursors.
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Abstract Hemopoietic cells develop in a complex milieu that is made up of diverse components, including stromal cells. Wnt genes, which are known to regulate the fate of the cells in a variety of tissues, are expressed in hemopoietic organs. However, their roles in hemopoiesis are not well characterized. In this study, we examined the roles of Wnt proteins in hemopoiesis using conditioned medium containing Wnt-3a. This conditioned medium dramatically reduced the production of B lineage cells and myeloid lineage cells, except for macrophages in the long-term bone marrow cultures grown on stromal cells, although the sensitivity to the conditioned medium differed, depending on the hemopoietic lineage. In contrast, the same conditioned medium did not affect the generation of B lineage or myeloid lineage cells in stromal cell-free conditions. These results suggested that Wnt proteins exert their effects through stromal cells. Indeed, these effects were mimicked by the expression of a stabilized form of β-catenin in stromal cells. In this study, we demonstrated that Wnt signaling regulates hemopoiesis through stromal cells with selectivity and different degrees of the effect, depending on the hemopoietic lineage in the hemopoietic microenvironment.
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