M2 macrophage generation medium dxf

DU145, 22RV1 and THP-1 cells were obtained from the Chinese Academy of Sciences Committee on Type Culture Collection Cell Bank (Shanghai, China). All cell lines were cultured in RPMI-1640 medium (Sigma-Aldrich; Merck, Darmstadt, Germany) supplemented with 10% fetal bovine serum and maintained in 5% CO₂ at 37 °C. These cell lines were authenticated by the Chinese Academy of Sciences Committee using short tandem repeat (STR) profiling. To obtain THP-1-derived M2 macrophages, THP-1 cells (2×10⁵ cells/well) were treated with 10 ng/ml phorbol 12-myristate 13-acetate (PMA) for 24 h and then cultured with IL-4 (25 ng/ml) and IL-13 (25 ng/ml) for another 48 h. Human PBMCs (peripheral blood mononuclear cells) were purchased from Shycbio (Shanghai, China) and stored in a -80 ℃ ultralow temperature freezer. For PBMC-derived M2 macrophages, an appropriate amount of complete M2 macrophage Generation Medium DXF (Promo Cell, Heidelberg, Germany) was incubated with the cells according to the manufacturer's instructions.

Human primary M2 macrophages (PromoCell) were cultivated with M2-Macrophage Generation Medium DXF (PromoCell). Cells were detached by Macrophage Detachment Solution DXF (PromoCell) following the manufacturer's protocol. After detaching, cells were filled immediately into Nutrimix bags (B. Braun Melsungen) (2 × 10⁶ cells in 10 mL medium each bag) for parabolic flight, or in serological pipettes (1 mL cell suspension with 0.25–0.5 × 10⁶/mL) for clinorotation experiment [18 (link), 19 (link)].