Glucocorticoid receptors and cortico‐sensitivity in a human clonal monocytic cell line, CM‐SM

Abstract

CM-SM is a clonal line of human precursor mononuclear phagocytes inducible to macrophage differentiation in response to the tumor promoter phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Untreated CM-SM cells contain single class, high-affinity (KD = 4.0 X 10(-9) M) glucocorticoid-specific receptor sites (approximately 60,000 per cell), as measured by a whole cell assay, at 37 degrees C, using [3H]triamcinolone acetonide (TA). Exposure of CM-SM to dexamethasone (DEX) produced a progressive, dose- and time-related series of changes in CM-SM cell growth, saturation density, morphology, and functional properties, with half-maximal effects at about 10(-9) M for DEX. TA-receptor sites rapidly decreased (about 70%) after DEX treatment, without any apparent change in steroid specificity and affinity. After 5 days in culture with a saturating concentration (3.6 X 10(-8) M) of hormone, the cells reached a saturation density of about 9.0 X 10(6) viable cells/ml (about 4.0 X 10(6) viable cells/ml in the controls), while the modal volume of the resulting cell population was approximately 60%, as compared to the volume of untreated cells. DEX-treated cells appeared less differentiated than controls, as assessed by combined morphologic, antigenic, and cytoenzymatic analyses. DEX almost completely inhibited TPA activation of the following macrophage functions: adherency to the culture plate, expression of lysosomal enzymes, Fc and C3 receptors, and stimulation of phagocytosis. After removal of DEX, the cells, within a few passages, returned to a state apparently identical to the untreated controls and could be induced to macrophage differentiation in response to TPA.

Glucocorticoid receptors and cell cycle progression in human melanoma cell lines

“…in good agreement with reports in the literature for glucocorticoid receptors in WI-38 human fibroblasts (Rosner and Cristofalo, 1981) and monocytes in vitro (Ranelletti et al, 1983). However, the numbers per cell were quite high compared to leukemic cells (Konior Yarbro et al, 1977;Kontula et al, 1980;Costlow et al, 1982).…”

“…We have investigated the effect of 1 h treatment with a pulse dose of a synthetic glucocorticoid on the cell cycle progression and proliferation of human melanoma cells in vitro. In four cell lines studied, the number of saturable glucocorticoid binding sites: 51,000 to 170,000 per cell (Table 3), were -5 -=" 100 in good agreement with reports in the literature for glucocorticoid receptors in WI-38 human fibroblasts (Rosner and Cristofalo, 1981) and monocytes in vitro (Ranelletti et al, 1983). However, the numbers per cell were quite high compared to leukemic cells (Konior Yarbro et al, 1977;Kontula et al, 1980;Costlow et al, 1982).…”

Epstein-Barr Virus DNA Sequences in Precursor Monocyte-Macrophage Cell Lines Established from the Bone Marrow of Children with Maturation Defects of Haematopoiesis

“…Four autonomous cell lines were established from bone marrow blood samples obtained from young patients affected by a variety of disorders: a 3 year-old male with congenital hypoplastic (DiamondBlackfan) anaemia (line CM-S), a 7 month-old male with cyclic neutropenia (line AD-S), a 1 year-old female also with cyclic neutropenia (line CV-S) and a 6 month-old male with cyclic dysmyelopoiesis associated with pancreatic insufficiency (Shwachman-Diamond syndrome) (line ROV-S). The establishment, growth properties, ultrastructural and in vitro differentiation characteristics of the four lines have been presented in detail (Monaco et al, 1982;Ranelletti et al, 1983;Bertolini et al, 1984;Sambuy et al, 1985;Revoltella et al, 1986;Romitti et al, 1986). Tables 1 and 2 summarize the observed i mmunotype and other various markers of the lines, Fig.…”

“…1 illustrates the typical appearance of the cell lines. Briefly, they can be characterized as monocyte-macrophage progenitor cell lines in that the cells grow in suspension in an undifferentiated form, but they can be stimulated to differentiate and resemble macrophage cells in the presence of appropriate chemicals, such as the tumour promoter 12-0-tetradeeanoyl phorbol 13-acetate (TPA) (Monaco et al, 1982;Ranelletti et al, 1983Ranelletti et al, , 1986Bertolini et al, 1984;Sambuy et al, 1985;Revoltella et al, 1986;Romitti et al, 1986). Criteria for monoeyte-macrophage differentiation in response to TPA included adherence to the culture vessel, loss of replicative capacity, positive non-specific esterase activity, production of colony-stimulating factors for granulocyte and macrophage precursors, interleukin 1 (IL-1) and lysozyme, expression of histocompatibility class II (HLA DR and DQ) antigens and monocyte-restricted membrane antigens, Fc and C3b rosette formation, immunophagocytosis and phagocytic properties.…”

Growth factors that stimulate human granulocyte—macrophage colony formation produced by the cell line CM‐S

“…In the presence of suitable chemical inducing agents such as some tumor promoter phorbol diesters, the cells become adherent, stop dividing, differentiate, and rapidly acquire morphologic and functional properties of the monocyte-macrophage lineage, including lysosomal enzymes, Fc and C, rosette formation, and the capacity to phagocytose inert particles or bacilli Ranelletti et al, 1983). The cells are maintained in Dulbecco's modified Eagle's medium (DMEM) (Grand Island Biological, New York) supplemented with 20% fetal calf serum (Flow Laboratories, Ayrshire, Scotland), 10% trypticase soy broth (BBL Microbiology Systems, Becton Dickinson, Cockeysville, MD), 100 Uiml penicillin, and 50 pg/ml streptomycin, cultured in tightly closed plastic flasks (T30, Falcon Division, Becton Dickinson, Oxnard, CA) at 37°C.…”