Composition and functional integrity of the in vitro hemopoietic microenvironment in acute myelogenous leukemia: effect of macrophage colony-stimulating factor

Abstract

In the present work, we have investigated the composition and hemopoietic supportive capacity of eleven normal and six acute myelogenous leukemia (AML) marrow-derived stromal adherent layers, established in the absence or in the presence of recombinant human colony-stimulating factor 1 (rhCSF-1, macrophage colony-stimulating factor). Two of six AML adherent layers were deficient in composition (i.e., no confluency, reduced numbers of macrophages and fibroblastic progenitors, and no fat cell formation), resulting in reduced CSF-1 production and a poor hemopoietic supportive capacity (assessed by the ability of an irradiated stroma to sustain the growth of myeloid, erythroid, and multipotential progenitors derived from a second innoculum of normal bone marrow). Four out of six AML adherent layers showed levels of macrophages, fibroblastic progenitors, fat cells, and CSF-1 similar to those observed in adherent layers from normal bone marrow; however, their capacity to sustain normal hemopoiesis was still significantly reduced. The deficient hemopoietic supportive capacity of all AML adherent layers correlated with the presence of a soluble activity in the culture supernatant that inhibited hemopoietic colony formation. Addition of rhCSF-1 during the establishment of AML adherent layers significantly increased their hemopoietic supportive capacity. In contrast, the hemopoietic supportive capacity of normal adherent layers was reduced by rhCSF-1. The opposite effects of rhCSF-1 on the hemopoietic supportive capacity of normal and AML adherent layers correlated with the levels of the soluble inhibitory activity, that is, increased levels in cultures containing rhCSF-1-treated normal adherent layers, and slightly reduced levels in cultures of rhCSF-1-treated AML layers. These results indicate that, despite a morphologically normal composition in most cases (four out of six), the hemopoietic microenvironment developed in long-term marrow culture (LTMC) from all AML marrows analyzed has a deficient hemopoietic supportive capacity, due, at least in part, to the production of hemopoietic inhibitor(s). Such a deficiency can be partially overcome by establishing the stroma layers in the presence of rhCSF-1.