Bioassay of human granulocyte colony-stimulating factor using human promyelocytic HL-60 cells.
A new method for an assay of human granulocyte colony-stimulating factor (hG-CSF) has been developed using human promyelocytic HL-60 cells. The proliferation of HL-60 cells had been suppressed by the addition of dimethyl sulfoxide (DMSO) or retinoic acid (RA). These differentiating agent-treated HL-60 cells exhibited an increase in their number in response to recombinant hG-CSF (rhG-CSF). Neither dibutyl-cAMP nor interferon-gamma (IFN-gamma)-treated HL-60 cells, however, showed an increase in their number in response to rhG-CSF. The proliferation rate of DMSO-pretreated HL-60 cells was linearly increased from 0.3 to 10 ng/ml of rhG-CSF. L-Value of HL-60 cells assay was 0.027 +/- 0.012. The activities of non-glycosylated rhG-CSF produced by Escherichia coli and glycosylated rhG-CSF produced by chinese hamster ovary (CHO) cells were compared using DMSO-treated HL-60 cells; no significant difference between them. DMSO-treated HL-60 cells also responded to interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF), but did not respond to erythropoietin or macrophage colony-stimulating factor, suggesting that the responsiveness of these cells to growth factor is restricted to myelogenic cytokines. In conclusion, DMSO- or RA-treated HL-60 cells are useful for the measurement of bioactivity of hG-CSF.
Yamaguchi
T
T
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Tokyo, Japan.
Yamaguchi
T
T
Kogi
M
M
Yamamoto
Y
Y
Hayakawa
T
T
D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D006224:Cricetinae; D016207:Cytokines / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D006031:Glycosylation; D016179:Granulocyte Colony-Stimulating Factor / Q000032:analysis*; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects*; D011994:Recombinant Proteins; D014212:Tretinoin / Q000494:pharmacology*
D016207:Cytokines; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D014212:Tretinoin; D004121:Dimethyl Sulfoxide