Established and Patient-Derived PDAC Cell Culture

Established PDAC cell lines, BxPC-3 and Panc-1, were obtained from American Type Culture Collection (Manassas, VA) and were grown in RPMI-1640 (Life Technologies, Grand Island, NY) and high glucose DMEM (Hyclone, Logan, UT), respectively. Low-passage patient-derived PDAC cells, 10.05, as well as CAFs were grown in high glucose DMEM without sodium pyruvate (Life Technologies)^{54 (link),87 (link)}. All medium was supplemented with 10% heat-inactivated fetal bovine serum (HI FBS; Life Technologies). Medium for BxPC-3 and Panc-1 was also supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin (Sigma Aldrich, St. Louis, MO), while 10.05 and CAFs were cultured in absence of antibiotics. Cells were maintained in a humidified environment of 5% CO₂ in air at 37 °C. All cells were passaged at 70–90% confluency; established PDAC lines and CAFs were used below passage 20. Patient-derived PDAC cells were used below passage 10, were authenticated by STR analysis (CellCheck with IDEXX BioResearch) and were tested regularly for mycoplasma contamination.