The human promyelocytic HL60 cell line: a model of myeloid cell differentiation using dimethylsulphoxide, phorbol ester and butyrate.
The release of the reactive oxygen species that accompanies the oxidative burst was studied in HL60 cells differentiated with either dimethylsulphoxide, butyrate or phorbol myristate acetate in order to establish the extent to which differentiated cells are phenotypically similar to human neutrophils, monocytes and macrophages. When phorbol myristate acetate was used as a stimulus, the rates of superoxide production by dimethylsulphoxide and butyrate differentiated HL60 cells was not significantly different from those observed in neutrophils and monocytes isolated from normal peripheral blood. Similar results were obtained when luminol-dependent chemiluminescence was measured in the presence of horseradish peroxidase using phorbol myristate acetate as the stimulus. However, in the absence of horseradish peroxidase, the luminol-dependent chemiluminescence in the dimethylsulphoxide and butyrate-differentiated HL60 cells was significantly lower than that of the control cells isolated from human blood, reflecting the absence of myeloperoxidase in the differentiated cells. In contrast, HL60 cells differentiated by phorbol myristate acetate failed to show any increased generation of superoxide or luminol-dependent chemiluminescence upon stimulation. Impaired release of lysosomal enzymes by the chemically differentiated cells suggests impairments in the extent of differentiation resulting in cells with defective azurophilic degranulation processes. It is concluded that HL60 cells differentiated by the above agents are somewhat controversial models of promyelocyte differentiation into typical neutrophilic, monocytic and macrophage-like cells.
Ahmed
N
N
Department of Biochemistry, Faculty of Science, Australian National University, Canberra ACT.
Williams
J F
JF
Weidemann
M J
MJ
D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002454:Cell Differentiation / Q000187:drug effects; D003574:Cytochrome c Group / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008163:Luminescent Measurements; D008247:Lysosomes / Q000201:enzymology; D008954:Models, Biological; D009504:Neutrophils / Q000378:metabolism; D009580:Nitroblue Tetrazolium; D010084:Oxidation-Reduction; D010641:Phenotype; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002087:Butyrates; D003574:Cytochrome c Group; D020148:Butyric Acid; D009580:Nitroblue Tetrazolium; D013755:Tetradecanoylphorbol Acetate; D004121:Dimethyl Sulfoxide