Ct26 colorectal carcinoma

DC 2.4 cells were collected from C57BL/6 mice and infected with GM-CSF-transfected medullary cells with myc/raf oncogenes, as previously described (27 (link)). CT26 colorectal carcinoma (American Type Culture Collection, Manassas, VA, USA) and DC 2.4 cell lines were cultivated in RPMI 1640 (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at 37°C in an incubator under humid conditions with 50 ml/l CO₂. C57BL/6 mice (n=6; male) aged 6–8 weeks (weight, 18–20 g) were obtained from the Chinese Academy of Science Shanghai Laboratory Animal Center (Shanghai, China). Animal experiments in this study were conducted in accordance with the internationally accepted principles for laboratory animal use and care. The study was approved by the Committee on the Ethics of Animal Experiments of China Medical University (Shenyang, China).

The 4T1 breast cancer, MC38 colon adenocarcinoma, CT26 colorectal carcinoma, and B16-F10 (B16) melanoma cell lines were purchased from the American Type Culture Collection. 4T1, MC38, and B16 cells were cultured in RPMI (Gibco) supplemented with 10% fetal bovine serum, 1% nonessential amino acids, 2 mM l-glutamine, 0.5% penicillin/streptomycin, and 55 µM 2-mercaptoethanol. These cell lines were authenticated by morphology, phenotype, and growth, and routinely screened for Mycoplasma, and were maintained at 37°C in a humidified 5% CO₂ atmosphere.