RESUMEN
Upon treatment with the phorbol ester, tetradecanoylphorbol 13-acetate (PMA), peripheral mononuclear blood cells from patients with acute myeloid leukemia secrete into serum-free cell-conditioned media (PMA-CCM) at least three distinct nondialysable 'hematopoietic' factors: granulocyte-colony-stimulating factor (G-CSF), granulocyte/macrophage-colony-stimulating factor (GM-CSF) and erythroid differentiation factor (EDF, activin A). G-CSF was identified by its stimulation of [3H]thymidine incorporation into a G-CSF-responsive cell line, NSF-60, and the inhibition of its stimulation by a G-CSF-specific monoclonal antibody (MAB). GM-CSF was identified by its stimulation of [3H]thymidine incorporation into a GM-CSF-responsive line, TALL-101, and the inhibition of its stimulation by a GM-CSF-specific MAB. EDF was identified by its ability to stimulate erythroid differentiation in mouse erythroleukemia cell lines, its identical retention times to those of authentic EDF on three successive reverse-phase HPLC columns and characterization of its penultimate N-terminal residue as leucine which is the same as that of authentic EDF. Both authentic EDF and the erythroid-stimulating activity in PMA-CCM were found to act synergistically with a suboptimal inducing concentration of a well-studied inducing agent, dimethyl sulfoxide, in inducing erythroid differentiation. In addition, a fourth activity was observed in PMA-CCM: normal human fetal bone marrow cell-proliferation stimulating activity (FBMC-PSA). FBMC-PSA was identified by its ability to stimulate the growth of granulocytes and macrophages in FBMC suspension cultures, which neither recombinant G-CSF or GM-CSF were found to do.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Inhibinas/metabolismo , Leucemia Mieloide/metabolismo , Acetato de Tetradecanoilforbol/farmacología , Activinas , Enfermedad Aguda , Médula Ósea/embriología , Medios de Cultivo , Sustancias de Crecimiento/metabolismo , Humanos , Células Tumorales CultivadasRESUMEN
An 8-21 fold multiplication of myeloid cells and macrophages was observed in tissue culture from human fetal bone marrow. Proliferation of the cells was triggered by medium conditioned by acute myelocytic leukemic cells exposed to 12-O-tetradecanoyl phorbol 13-acetate (TPA). The medium was designated as TPA-treated cell conditioned medium (TPACCM). The cycle of events in fetal bone marrow cell culture began with a sharp decrease in the total number of cells. At this juncture a predominance of primitive macrophage-like cells positive for macrophage markers was present in the culture. The process of multiplication began with rapid proliferation of promyelocytes. Simultaneous proliferation of granulocyte-macrophage (GM) colony forming cells (CFC) indicated that at least some of the promyelocytes might act like CFC. The absolute number of committed CFC then increased 5-11 fold, the number that approximated a total multiplication of FBM cells. The proliferation continued with the culture containing some blast cells and showing simultaneous differentiation of the cells into mature granulocytes and macrophages. The cells with macrophage markers persisted throughout the culture period. To determine more precise definition of the role of primitive macrophages and promyelocytes in FBM cell multiplication, experiments with purified fractions of these cells have to be done. TPACCM purification and isolation of active substances is also suggested. These investigations might result in obtaining a pool of BM cells in vitro suitable for BM transplantation.
Asunto(s)
Células de la Médula Ósea , Células Madre Hematopoyéticas/citología , Forboles/farmacología , Acetato de Tetradecanoilforbol/farmacología , Médula Ósea/embriología , División Celular , Ensayo de Unidades Formadoras de Colonias , Medios de Cultivo , Técnicas de Cultivo , Feto/citología , Granulocitos/citología , Humanos , Leucemia Mieloide Aguda , Macrófagos/citologíaRESUMEN
The capacity of fetal bone marrow to form stromal cell colonies, granulocyte-macrophage colonies, stromal cell monolayers and to produce granulocyte-macrophage colony stimulating activity from these monolayers was evaluated in comparison to adult bone marrow. Granulocyte-macrophage colony formation on routine feeder layers was approximately equal in both cases. The fetal bone marrow colonies were larger in size than those from adult precursors. There were at least ten times more stromal cell precursors in fetal bone marrow than in adult bone marrow. The fetal bone marrow stromal cell monolayer formed within 7-12 days whereas adult stromal monolayer formation required 3-4 weeks. Fetal bone marrow stromal cell monolayer produced granulocyte-macrophage colony-stimulating activity (GM-CSA) effective only for fetal GM precursors, whereas adult bone marrow produces GM-CSA effective for fetal and adult normal and CML bone marrow.