Functional Differentiation of HL-60 Cells by Dimethylsulfoxide and Phorbol 12-myristate 13-acetate

ABSTRACT

PURPOSE: During hematopoietic differentiation of HL-60 cells by DMSO and PMA, we demonstrated functional changes of HL-60 cells-phagocytic activity, respiratory burst, antibody-dependent cell-mediated cytotoxicity(ADCC) and opsonophagocytic activity. METHODS: HL-60 cells(ATCC CCL-240), cultured in RPMI 1640 and supplemented with 10% FBS, were induced to differentiate by adding 1.0% DMSO or 16nM PMA. On the 4th and 7th day after stimulation as well as before stimulation, the cells were analyzed for functional differentiation. Phagocytic activity was measured by flow cytometry after incubation of the cells with fluorochrome-conjugated beads. Respiratory burst was measured by chemiluminescence assay. ADCC was measured by hemoglobin release assay. Opsonophagocytic activity was measured by fungicidal assay using Candida albicans. RESULTS: Phagocytic activity of HL-60 cells was not increased by differentiation with DMSO. But PMA induced increase of phagocytic activity on 7th day. Respiratory burst studied by chemiluminescence was increased up to 4-fold on 7th day by DMSO. PMA induced increase upto 3-fold on 4th day. ADCC was increased upto 3-fold on 4th day by DMSO, but PMA induced little increase in ADCC. Opsonophagocytic activity was increased upto 3-fold on 4th day by DMSO or PMA. On differentiation with DMSO, respiratory burst correlates with FcgammaRI and FcgammaRII. ADCC and opsonophagocytic activity correlate with CD11b/CD18. On differentiation with PMA, phagocytic activity correlates with FcgammaRII. Respiratory burst correlates with CD11b. ADCC and opsonophagocytic activity of HL-60 cells correlate with CD18. CONCLUSION: Treatment of HL-60 cells with DMSO or PMA induces functional maturation differently. Phagocytic activity, respiratory burst, ADCC and opsonophagocytic activity of HL-60 cells correlated with expresson of FcgammaR and Mac-1.