Production of human granulocyte colony stimulating factor by various kinds of stromal cells in vitro detected by enzyme immunoassay and in situ hybridization

Abstract

Abstract. Production of human granulocyte colony stimulating factor (G-CSF) by stromal cells was studied in vitro. Induction of G-CSF by interleukin 1 (IL-1) and lipopolysaccharide (LPS) was compared using enzyme immunoassay in various kinds of stromal cells. Primary human bone marrow stromal cells, a human bone marrow-derived stromal cell line (KM-102), and peripheral blood monocytes secreted small amounts of G-CSF without stimulation, while vascular endothelid cells and skin fibroblasts secreted G-CSF only when induced by IL-1 or LPS. The production of G-CSF by monocytes was stimulated predominantly by LPS, whereas that by KM-102 cells, endothelial cells, and fibroblasts was induced by IL-1 but much less so by LPS. IL-1 and LPS stimulated similar levels of G-CSF production by primary bone marrow strom d cells which consisted of various types of cells. In situ hybridization for G-CSF mRNA showed that only a small proportion of primary bone marrow stromal cells expressed a large amount of G-CSF mRNA upon stimulation. The positive cells were round or oval in shape, while most of the spindle-shaped stromal cells were negative for specific grains. Although further characterization of positive

Cytokines secreted by bone marrow stromal cells protect c-KIT mutant AML cells from c-KIT inhibitor-induced apoptosis

“…As seen in Figure 2E, over 80% of Kasumi-1 cells exhibited cell surface expression of the G-CSF receptor compared to <20% of SKNO-1 and HMC1.1 cells. In agreement, it has been previously shown that Kasumi-1 cells express significantly more G-CSF receptor mRNA than SKNO-1 cells 11 . These data suggest that the cell surface expression of the G-CSF receptor is correlated with the capacity of the cytokine to reverse sensitivity to c-KIT inhibition in Kasumi-1 cells and its failure to do so in SKNO-1 and HMC1.1 cells.…”

G-CSF Administration Accelerates Cutaneous Wound Healing Accompanied With Increased Pro-Hyp Production In db/db Mice

“…It has also been reported that inflammation-related products, such as IL-1 and lipopolysaccharide, stimulate G-CSF production by fibroblasts originating from the skin [27]. In this study, we showed that G-CSF levels in wounded tissue reached a plateau as early as 3 hours after total skin excision, and spindle-to ovalshaped mesenchymal cells (probably fibroblasts) in the wounded tissue express G-CSF.…”

Macrophage Depletion Disrupts Immune Balance and Energy Homeostasis

“…Therefore, we hypothesized that macrophages directly regulate G-CSF expression. We used OP9 bone-marrow stromal cells, which have been known to secrete G-CSF especially after stimulation by either lipopolysaccharide or IL-1 [33], [34], [35]. To study whether macrophages regulate G-CSF expression through secreted cytokines, we treated OP9 cells with conditioned media from either bone-marrow or peritoneal derived macrophages of wild type mice.…”