Glioma Cell Lines and Macrophage Cultivation

Glioma cell lines U87 and human mononuclear macrophage line (THP-1) were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human microglia line HMC3 and murine glioma cell line GL261 were obtained from American Type Culture Collection (Manassas, VA, USA). The extraction of the patient-derived primary glioma cells (PGC28) were extracted as previously described.^{21 (link)} U87 and GL261 were maintained in Dulbecco’s modified Eagle’s medium (DMEM, 10566024, Gibco) supplemented with 10% fetal bovine serum (FBS, 16140071, Gibco) and 1% penicillin/streptomycin (10378016, Gibco). PGC28 and THP-1 were maintained in RPMI-1640 medium (61870036, Gibco) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. THP-1 monocytes were primed with 5 nM PMA (P1585, Sigma) for 48 hours to become THP1-derived macrophages. HMC3 were maintained in Minimum Essential Medium (MEM, 12561056, Gibco) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. All cells were cultured at 37°C with 5% CO₂.