*5.5. Human THP-1 Pre-Monocyte Culture*

The non-adherent human pre-monocyte of the THP-1 cell lineage was acquired from the Rio de Janeiro Cell Bank (Rio de Janeiro, RJ, Brazil, cat.: BCRJ: 0234) and cultured according to the manufacture instructions. The suspension cells were cultured in 75 cm3 culture flasks (Corning Inc., New York, NY, USA) with RPMI-1640 medium (Gibco, Invitrogen Corp., Waltham, MA, USA) supplemented with 23 mM NaHCO3, 13 mM C6H12O6, 10 mM Hepes, 2 mM L-glutamine, 1 mM sodium pyruvate, 10% FBS (Fetal Bovine Serum, Cultilab, São Paulo, SP, Brazil), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Gibco, Invitrogen Corp., Waltham, MA, USA). The cell was maintained at 37 ◦C in an atmosphere containing 5% CO2 at a density of 2–8 × <sup>10</sup><sup>5</sup> cells/mL, not exceeding the concentration of 1 × 106 cells/mL. Total cell number and cell viability were periodically monitored in a Neubauer chamber by Trypan blue exclusion (Trypan blue 0.4% in PBS pH 7.2; 1:1; *v*/*v*).
