Pancreatic Cancer Cell Line Cultivation

PANC-1, CAPAN-1, BxPC-3, and AsPC-1 pancreatic cancer cell lines were purchased from American Type Culture Collection (ATCC) (Manassas, USA). Cell lines were selected based on their variable MUC1-CE expression, ranging from strong (PANC-1), moderate (CAPAN-1), moderate-weak (BxPC-3) and weak (AsPC-1) levels (Hull et al. 2020 (link)). PANC-1 cells were cultured in Dulbecco’s Modified Eagle Medium supplemented with 10% foetal calf serum (FCS) and 1% penicillin/streptomycin (P/S). CAPAN-1 cells were cultured in Iscove’s Modified Dulbecco’s Medium supplemented with 20% FCS and 1% P/S. BxPC-3 and AsPC-1 cells were cultured using Roswell Park Memorial Institute 1640 medium supplemented with 10% FCS and 1% P/S. For all experiments, cells were grown to confluence in T75 flasks in a 5% carbon dioxide in air environment set to 37 °C. At confluence, cells were washed twice with DPBS and detached from the flasks using TrypLE™ Select Enzyme (1X) (Thermo Fisher Scientific Australia Pty Ltd, Scoresby, Australia). All cell lines were used within three months of resuscitation.