Induction and Identification of Macrophage Polarization

THP-1 cells and the culture medium were purchased from Procell (Wuhan, China; 1×10⁶cells, third generation; Identified through STR; No Mycoplasma contamination detected). THP-1 cells were inoculated in medium and cultured at 37 °C, 5% CO2, and 90% humidity in an incubator. THP-1 cells were seeded in 6-well plates, and M0 macrophages were stimulated with 100 ng/mL phorbol 12-myristate 13-acetate (PMA, Solarbio, China) for 48 h. M0 macrophages were incubated with 20 ng/mL IL-4 and 20 ng/mL IL-13 (Solarbio) for 48 h to induce M2 polarization. Macrophages were collected, and subsequently subjected to qRT-PCR analysis to detect CD11b, CD14, CD68, iNOS, IL-10, and CD206 to identify M0 and M2 macrophages (Table 1 for primer sequences).

Primers Used in This Manuscript