Establishing Hypoxic Bladder Cancer Cell Lines

Cell lines in this study include six human BCa lines (T24, UMUC3, J82, 5637, BIU87, and RT4), a human bladder epithelial cell line (SV-HUC-1), and a human embryonic kidney cell line (293 T). All cell lines were provided by the Stem Cell Bank, Chinese Academy of Sciences in Shanghai and have recently been authenticated. RPMI-1640 medium was used for SV-HUV-1, T24, 5637, BIU87 and RT4 cell culture, DMEM medium was applied for UMUC3 and 293 T cell culture, and MEM medium was utilized for J82 cell culture. All culture mediums were freshly prepared with 10% fetal bovine serum (FBS). To simulate the hypoxic conditions, cells were subjected to a 4-h pretreatment of CoCl2 (100 μM, Sigma c8661) prior to collection [36 (link), 37 (link)].