Murine Pancreatic Tumor Model Establishment

The “FC1242” (C57BL/6 background) murine PDA cell line was isolated from the pancreatic tumor of a KPC mouse (pdx1^cre/+;KRas^LSL-G12D/⁺; p53^R172H/+) as we described previously^{51 (link)}. In select experiments, we utilized FC1242 KPC-derived tumor cells (1 × 10⁶), which we engineered to express OVA using pCI-neo-cOVA (gift of Maria Castro; Addgene plasmid # 25097) as we described^{21 (link)}. To establish orthotopic pancreatic lesions in vivo, tumor cells (10⁵) were suspended in phosphate-buffered saline (PBS) with Matrigel (BD) in a 1:1 ratio and were injected into the body of the pancreas via laparotomy. In select experiments, DC (2.5 × 10⁴) were admixed with tumor cells and co-transferred to pancreata of mice. In other experiments, OTII T cells (5 × 10⁴) were admixed with equal numbers of FC1242 and FC1242.Ova tumor cells (2.5 × 10⁴ each) and administered subcutaneously. For our liver metastases model, PDA cells (1 × 10⁶) were administered into the portal venous system via direct splenic injection followed by splenectomy, as we reported^{52 (link)}.