Murine Colon and Lung Cancer Cell Cultures

Cells of the murine colon carcinoma cell line CT26 (harboring a ras mutation but wild type for p53 [14 (link)]) and the Lewis lung carcinoma cell line LLC (harboring mutations in both ras and p53 [15 (link), 16 (link)]) were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, USA) and the Riken BioResource Center (Ibaraki, Japan), respectively. Both lines were cultured in RPMI1640 medium (Sigma-Aldrich Inc., Tokyo, Japan) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Tokyo, Japan), 100 mg/mL streptomycin and 100 U/mL penicillin (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan). Cells were incubated in a humidified atmosphere at 37°C with a 5% CO₂. For subculturing, cells were rinsed with Ca^2 + − and Mg²⁺ − free phosphate-buffered saline (PBS; FUJIFILM Wako) and dispersed with 0.25% trypsin containing 0.5 mM ethylenediaminetetraacetate (FUJIFILM Wako). The number of cells were counted with a Countess II FL (Thermo Fisher Scientific) or a Scepter 2.0 (Merck KGaA, Darmstadt, Germany).