Orthotopic Pancreatic Tumor Model in Mice

Animal protocols were approved by the University of Colorado Animal Care and Use Committee. Eight weeks old C57BL/6 and Nlrp3^−/− male mice were purchased from Jackson Laboratories. Eight weeks old NCG male mice were purchased from Charles Rivers. For orthotopic pancreatic tumor challenge, mice received an intrapancreatic injection of FC1242 tumor cells derived from KPC mice backcrossed with C57BL/6 background (14 (link)). Cells were kindly provided by Dr. S.D. Karam through an agreement with Dr. Tuveson, who generated them. FC1242 cells were plated for 2 days in RPMI1640 (Corning) with 10% (v/v) FBS (Corning) and 1% penicillin-streptavidin (Corning). Cells were collected and suspended in 1:1 solution with RPMI1640 medium containing 1% penicillin-streptavidin and Matrigel (Corning). A total of 1 × 10⁵ cells in 20 µL were injected into the body of the pancreas via laparotomy. Mice were sacrificed at 14 or 21 days after the surgery, and tumor dimensions and weight were recorded. Tumor volumes were calculated as described previously (15 (link)). For survival, 2 × 10⁵ cells were injected following the same protocol. Natural death and humane endpoints were applied to collect datapoints following the guidelines from the University of Colorado (Aurora, CO).