Primary 83,320 KPC cells have previously been isolated from end-stage Pdx1-Cre, LSL-KrasG12D/+, LSL-Trp53R172H/+ KPC mice (27 (link), 29 (link), 35 (link)). Telomerase-immortalized fibroblasts (TIFs) were also generated previously (18 (link), 28 (link), 76 (link)). KPC cells, TIFs, and human embryonic kidney (HEK) 239T cells were maintained in Dulbecco’s modified Eagle’s media (DMEM; high glucose, pyruvate; Gibco) supplemented with 10% fetal bovine serum (FBS; HyClone) and 10 mM Hepes (Gibco). TKCC05 cells were maintained in media consisting of DMEM/Ham’s F12 media (Gibco) supplemented with 7.5% FBS, 10 mM Hepes, 13.3 mM glucose, hydrocortisone (40 ng/ml), insulin (0.1 IU/ml), and human recombinant epidermal growth factor (EGF; 10 ng/ml). TKCC10 cells were maintained in 1:1 M199 media/Ham’s F12 medium (Gibco) supplemented with 7.5% FBS, 15 mM Hepes, 2 mM glutamine, 1× MEM vitamins, apotransferrin (25 ng/ml), insulin (0.2 IU/ml), 6.5 mM glucose, hydrocortisone (40 ng/ml), EGF (20 ng/ml), triiodothyronine (0.5 pg/ml), and O-phosphoryl ethanolamine (2 μg/ml). All cells were cultured in the presence of penicillin-streptomycin (100 U/ml and 100 mg/ml, respectively) and maintained at 37°C and 5% CO2. Experiments were conducted at 20% oxygen in a Heracell 150i CO2/O2 incubator for KPC, TIF, and TKCC05 cell lines and at 5% oxygen for TKCC10 lines. All cell lines were mycoplasma-free.
Murphy K.J., Reed D.A., Vennin C., Conway J.R., Nobis M., Yin J.X., Chambers C.R., Pereira B.A., Lee V., Filipe E.C., Trpceski M., Ritchie S., Lucas M.C., Warren S.C., Skhinas J.N., Magenau A., Metcalf X.L., Stoehr J., Major G., Parkin A., Bidanel R., Lyons R.J., Zaratzian A., Tayao M., Da Silva A., Abdulkhalek L., Gill A.J., Johns A.L., Biankin A.V., Samra J., Grimmond S.M., Chou A., Goetz J.G., Samuel M.S., Lyons J.G., Burgess A., Caldon C.E., Horvath L.G., Daly R.J., Gadegaard N., Wang Y., Sansom O.J., Morton J.P., Cox T.R., Pajic M., Herrmann D, & Timpson P. (2021). Intravital imaging technology guides FAK-mediated priming in pancreatic cancer precision medicine according to Merlin status. Science Advances, 7(40), eabh0363.
Other organizations :
UNSW Sydney, The Kinghorn Cancer Centre, Garvan Institute of Medical Research, The Netherlands Cancer Institute, Oncode Institute, Turku Centre for Computer Science, University of Turku, Åbo Akademi University, University of Sydney, Royal North Shore Hospital, Glasgow Royal Infirmary, University of Glasgow, Victorian Comprehensive Cancer Centre, University of Melbourne, Inserm, Université de Strasbourg, University of South Australia, Centre for Cancer Biology, South Australia Pathology, University of Adelaide, Centenary Institute, Royal Prince Alfred Hospital, Anzac Research Institute, Chris O’Brien Lifehouse, Monash University, University of California, San Diego, La Jolla Bioengineering Institute
Culture media: Dulbecco's modified Eagle's media (DMEM; high glucose, pyruvate; Gibco) supplemented with 10% fetal bovine serum (FBS; HyClone) and 10 mM Hepes (Gibco) for KPC cells, TIFs, and human embryonic kidney (HEK) 239T cells
TKCC05 cells maintained in DMEM/Ham's F12 media (Gibco) supplemented with 7.5% FBS, 10 mM Hepes, 13.3 mM glucose, hydrocortisone (40 ng/ml), insulin (0.1 IU/ml), and human recombinant epidermal growth factor (EGF; 10 ng/ml)
TKCC10 cells maintained in 1:1 M199 media/Ham's F12 medium (Gibco) supplemented with 7.5% FBS, 15 mM Hepes, 2 mM glutamine, 1× MEM vitamins, apotransferrin (25 ng/ml), insulin (0.2 IU/ml), 6.5 mM glucose, hydrocortisone (40 ng/ml), EGF (20 ng/ml), triiodothyronine (0.5 pg/ml), and O-phosphoryl ethanolamine (2 μg/ml)
All cells cultured in the presence of penicillin-streptomycin (100 U/ml and 100 mg/ml, respectively) and maintained at 37°C and 5% CO2
Experiments conducted at 20% oxygen for KPC, TIF, and TKCC05 cell lines and at 5% oxygen for TKCC10 lines
All cell lines were mycoplasma-free
positive controls
None specified
negative controls
None specified
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