Thp1 cell line

MC38 cell line was obtained from Cell Resource Center of the Institutes of Biomedical Sciences at Fudan University (Shanghai, China) and maintained in the appropriate culture medium suggested by suppliers. THP1 cell line was obtained from Shanghai Institute of Cell Biology (Shanghai, China) and cultured with RPIM 1640 (Gibco, Grand Island, NY, USA) containing 10% FBS in 5% CO₂ at 37 °C.
Bone marrow cells were flushed out from the femurs and tibias of female C57BL/6 mice. After centrifugation for 5 min at 300×g, erythrocytes were eliminated and the remaining cells were cultured with DMEM supplemented with 10% fetal bovine serum and 10 ng/mL M-CSF (Peprotech, Rock Hill, NJ, USA; Cat. #315–02). Culture fluid was exchanged with culture medium every 3 days. Under these conditions, adherent macrophages were obtained within 7 days. Cells were harvested and seeded on 12-well plates without M-CSF for 6 h, then the cells were used for the experiments as BMDMs.

The bone marrow-derived macrophages (BMDMs) were generated from C57BL/6 mice as previously described, with minor modification. In brief, the tibiae and femurs were flushed with PBS for three times. After centrifugation at 1600 rpm (5424R, Eppendorf) for 5 min, the cells were cultured in RPMI 1640 medium supplemented with 10% FBS and 20 ng/mL rmM-CSF. The culture medium was exchanged every 3 days, and the adherent macrophages were obtained at 7 days.
The human monocytic THP-1 cell line was purchased from the Shanghai Institute of Cell Biology (Shanghai, China) and cultured in a humidified atmosphere with 5% CO₂ at 37 °C in RPMI 1640 medium supplemented with 10% FBS.