For the in vitro model, we used the HL-60 (ATCC® CCL-240™, Manassas, VA, USA) cell line from a human’s promyelocytic leukemia [83 (link)]. These cells were first cultured in suspension in complete culture medium (CCM) consisting of RPMI 1640 culture medium (Biowest, Nuaillé, France) containing 10% fetal bovine serum (Biowest, Nuaillé, France) and 1% penicillin/streptomycin (GIBCO, Invitrogen, Paisley, UK) into an incubator at 37 °C in 5% CO2.
Once the cells were growing at an exponential rate, they were differentiated into macrophage-like cells by culturing them in 96-well plates at 2 × 105 cells/well in 100 μL of CCM in the presence of 0.1 μg mL−1 phorbol myristate acetate (PMA) for 24 h. The cells were then reconditioned by changing the previous medium for 100 µL of fresh CCM, and once again incubated under the same conditions for another 24 h.
Once the cells were growing at an exponential rate, they were differentiated into macrophage-like cells by culturing them in 96-well plates at 2 × 105 cells/well in 100 μL of CCM in the presence of 0.1 μg mL−1 phorbol myristate acetate (PMA) for 24 h. The cells were then reconditioned by changing the previous medium for 100 µL of fresh CCM, and once again incubated under the same conditions for another 24 h.
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