Effect of recombinant and purified human haematopoietic growth factors on in vitro colony formation by enriched populations of human megakaryocyte progenitor cells
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Nonadherent low density T‐lymphocyte depleted (NALT ‐ ) marrow cells from normal donors were sorted on a Coulter Epics 753 Dye Laser System using Texas Red labelled My 10 and phycoerythrin conjugated anti HLA‐DR monoclonal antibodies in order to obtain enriched populations of colony forming unit‐megakaryocyte (CFU‐MK). The CFU‐MK cloning efficiency (CE) was 1.1 ± 0.5% for cells expressing both high densities of My 10 and low densities of HLA‐DR (My 10 +++ DR + ). This procedure resulted in an 18‐fold increase in CE over NALT ‐ cells. The effect of purified or recombinant human haematopoietic growth factors including erythropoietin (Epo), thrombocytopoiesis stimulating factor (TSF), interleukin 1α (IL‐1α), granulocyte colony stimulating factor (G‐CSF), granulocyte‐macrophage colony stimulating factor (GM‐CSF), macrophage colony stimulating factor (M‐CSF or CSF‐1) and interleukin‐3 (IL‐3) on MK colony formation by My10 +++ DR + cells was determined utilizing a serum depleted assay system. Neither Epo, TSF, CSF‐1, IL‐lα nor G‐CSF alone augmented MK colony formation above baseline (2.5 ± 0.8/5 × 10 3 My 10 +++ DR + cells plated). In contrast, the addition of GM‐CSF and IL‐3 each increased both CFU‐MK colony formation and the size of colonies with maximal stimulation occurring following the addition of 200 units/ml of IL‐3 and 25 units/ml of GM‐CSF. At maximal concentration, IL‐3 had a greater ability to promote megakaryocyte colony formation than GM‐CSF. The stimulatory effects of GM‐CSF and IL‐3 were also additive in that the effects of a combination of the two factors approximated the sum of colony formation in the presence of each factor alone. The CFU‐MK appears, therefore, to express HPCA‐1 and HLA‐DR antigens. These studies also indicate that GM‐CSF and IL‐3 are important in vitro regulators of megakaryocytopoiesis, and that these growth factors are not dependent on the presence of large numbers of macrophages or T cells for their activity since the My 10 +++ DR + cells are largely devoid of these accessory cells.Keywords:
Interleukin 3
Colony-stimulating factor
Colony-forming unit
Hematopoietic growth factor
Hematopoietic growth factor
Colony-stimulating factor
Monocyte
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Objective To investigate the effect of rhM CSF on hematopoietic recovery of chemotherapeutical lesions. Methods 100mg/kg CTX were ip injected into mice for 3 consecutive days to make the model of chemotherapeutical lesions. Mice were also ip injected with rhM CSF at doses of 0 1 2 0μg/day following the injection of CTX. Results 1 Observation of the peripheral hematological changes showed: The peripheral white blood cell count of treatment group(0 5 2 0 μg/day rhM CSF) was significantly higher than that of the control group at day 8 ( P 0 01). Monocyte and granulocyte counts of treatment group(0 1 2 0 μg/day rhM CSF) were significantly higher than those of the control group at day 8( P 0 01). Among which, 1 0 μg/day rhM CSF was most active. 2 On day 7, the bone marrow CFU GM yield of control group was significantly lower than that of treatment group, and no evident difference was found between the effect of rhM CSF and rhM CSF plus G CSF as stimulating factor. Conclusion rhM CSF could accelerate hematopoietic recovery in chemotherapeutically lesioned mice.
Hematopoietic growth factor
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Monocyte
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INTRODUCTION The macrophage colony-stimulating-factor (M-CSF or CSF-1) was initially described as an hematopoietic growth factor required for the survival, proliferation, differentiation and a variety of other functions of cells of the monocyte/macrophage lineage [1-4]. It also synergizes with others cytokines to induce the proliferation of earlier hematopoietic progenitors [5, 6]. More recently, a role of M-CSF in immunological [...]
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Monocyte
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Interleukin 3
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The effect of recombinant human growth factors: rhIL-6, rhGM-CSF and rhEPO on normal human hematopoietic progenitor cells was studied by using liquid and semisolid culture systems. The results showed that the expanding folds of progenitor cells after culturing with the three growth factors were 4.3 +/- 0.6, 9.4 +/- 0.9 and 13.7 +/- 1.0 respectively, being markedly higher than those of control (2.4 +/- 0.7, P < 0.01. In the presence of IL-6 alone, no CFU-E colony was observed and only CFU-GM colonies were formed in the semisolid culture. When IL-6 was added with EPO, the number of CFU-E colonies was significantly higher than that of EPO alone (P < 0.01). In the presence of IL-6 plus GM-CSF or IL-6 plus GM-CSF and EPO, CFU-Mix colonies were observed. Moreover, the total number of colonies was significantly higher than that of IL-6 or GM-CSF alone (P < 0.01). It is suggested that IL-6 is an important hematopoietic regulator. IL-6 may act on hematopoietic progenitor cells to enhance their proliferation. The target cells of IL-6 are the same as those of GM-CSF. IL-6 has synergistic action with EPO and GM-CSF, to enhance erythropoiesis with EPO and proliferation of multipotential progenitor cells with rhGM-CSF and EPO. However, the mechanism of the synergism between IL-6 and other growth factors is still unknown.
Interleukin 3
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Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a well-characterized hematopoietic growth factor. Recently, using purified recombinant-derived material, we have found that GM-CSF is also a potent activator of mature functional macrophages. Thus, we have found that exogenous GM-CSF augments the primary plaque-forming response to sheep red blood cells and that this effect is due to upregulation of la antigen expression and interleukin 1 production by the macrophages. We also show that GM-CSF inhibits the replication of <i>Trypanosoma cruzi </i>in cultured peritoneal macrophages and causes an accelarated clearance of <i>Salmonella typhimurium </i>from the peritoneal cavity of mice. These data indicate that GM-CSF is a multifunctional molecule stimulating both hematopoiesis and mature macrophage function.
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