Generation and Characterization of Murine Pancreatic Tumor Cell Lines

The generation of murine pancreatic tumor cell lines KPDC253, KPDC145 and KPDC143 was described previously^{34 (link)}. Briefly, a ~ 10 mm³ tumor piece was isolated at necropsy from mice with genetically-induced pancreatic ductal adenocarcinoma. The tumor was mechanically and enzymatically digested with 2 mg/mL Collagenase (MP Biomedical) in DMEM supplemented with 10% FBS for 30 min at 37 °C. All tumor tissues were cultured overnight in DMEM supplemented with 10% FBS. Adherent cells were isolated, serially diluted, and plated in 96-well plates. Single cell clones were identified and expanded and the Drp1 genotype was confirmed by PCR. In accordance with institutional guidelines, all mice used in the generation of these cell lines were monitored and euthanized when they reached pre-determined endpoints or exhibited features associated with disease such as weight loss. All animal studies and procedures were approved by the University of Virginia Institutional Animal Care and Use Committee.