Diverse Pancreatic Cancer Cell Lines

Pa01C, Pa02C, Pa04C, Pa14C and Pa16C cell lines were provided by A. Maitra (MD Anderson Cancer Center). The remaining PDAC cell lines and HEK-293T line were obtained from American Type Culture Collection (ATCC) and were maintained in either DMEM or RPMI 1640 supplemented with 10% FBS. Cell lines 12282, 14837 and 192 were derived from the doxycycline-inducible iKRAS mouse model^{3 (link)} and cultured in RPMI 1640 supplemented with 10% FBS. The RIE-1 cell line was received from R. Coffey (Vanderbilt University) and cultured in RPMI 1640 supplemented with 10% FBS. All cell lines were maintained in a humidified chamber with 5% CO₂ at 37 °C. Organoids were cultured at 37 °C in 5% CO₂. Cells were seeded in growth factor–reduced Matrigel (Corning) domes and fed with complete human feeding medium: advanced DMEM/F12-based WRN condition medium (L-WRN, ATCC CRL-3276), 1× B27 supplement, 10 mM HEPES, 0.01 µM GlutaMAX, 10 mM nicotinamide, 1.25 mM N-acetylcysteine, 50 ng ml^–1 hEGF, 100 ng ml^–1 hFGF10, 0.01 µM hGastrin I, 500 nM A83–01, 1 µM PGE2 and 10.5 µM Y27632. All cell lines were regularly monitored for mycoplasma contamination and immediately discarded if a positive result was detected. All cell line identities were verified by short-tandem-repeat profiling.