Differentiation of THP-1 monocytes into macrophages

THP-1 macrophage differentiation. THP-1 cells (5 × 10⁵ cells/well) were seeded in 24-well plates in RPMI 1640 supplemented with 10% FBS and 1% penicillin-streptomycin. After 24 h, cells were treated with PMA (50 ng/mL) for 24 h to induce differentiation. Cells were then washed with PBS to remove PMA and cultured for an additional 24 h in PMA-free complete medium to allow recovery/resting. Differentiation into macrophage-like cells was confirmed by flow-cytometric assessment of CD68 expression.