Pa01C, Pa02C, Pa03C, Pa04C, Pa14C and Pa16C cell lines were provided by A. Maitra (MD Anderson Cancer Center). The remaining PDAC cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were maintained in recommended media (DMEM or RPMI 1640 supplemented with 10% FBS and penicillin/streptomycin). Previously established organoids derived from pancreatic cancer were provided by David Tuveson (Cold Spring Harbor Laboratory). No patient samples were used in this study. All cell lines were maintained in a humidified chamber with 5% CO2 at 37°C. Pancreatic organoid 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. Colorectal cancer organoids were maintained as described (16 (link)). All cell lines were validated using short-tandem-repeat (STR) profiling and confirmed regularly as mycoplasma-negative.
Pdac cell lines
Manufactured by American Type Culture Collection
Sourced in United States
PDAC cell lines are a collection of human pancreatic ductal adenocarcinoma (PDAC) cell lines available from the American Type Culture Collection (ATCC). PDAC cell lines are derived from primary tumor samples and are used for research purposes.
Automatically generated - may contain errors
3 protocols using pdac cell lines
1
Establishing Pancreatic and Colorectal Cancer Organoids
2
PDAC Cell Line Cultivation and Authentication
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PDAC cell lines were purchased from the American Type Culture Collection. Cells were regularly tested negative for Mycoplasma and authenticated through Short Tandem Repeat (STR) analysis. All cancer cells were cultured in Dulbecco’s Modified Eagle Media with 10% fetal bovine serum (ThermoFisher Scientific, UK), at 37 °C with 5% carbon dioxide.
3
Validating NanoVelcro Chips for Pancreatic Cancer
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The performance of NanoVelcro Chips has previously been demonstrated and validated for several types of cancer, including prostate cancer and melanoma (Hou et al, 2013 (link)). However, we first sought to validate both the NanoVelcro and NanoVelcro/LCM chips, as well as our single-cell analysis, for pancreatic cancer using 4 PDAC cell lines obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) with various cellular phenotypes (Supplementary Table S1 ). Details of cell culture and calibration are available in the Supplementary Methods . In brief, PDAC cells were spiked in healthy donor blood to create artificial blood samples, and run using the method outlined below and in the Supplementary Methods . For initial calibration, 200 cells from each cell line were spiked into 1-ml healthy donor blood and were processed on the NanoVelcro platform at various flow rates in triplicate to determine the optimum flow rate for capture efficiency (Supplementary Figure S1A ). A flow rate of 1.0 ml h−1 was found to be the optimum flow rate, and was subsequently used for all patient samples (Supplementary Figure S1A ). Using the same method, capture efficiencies for each of the 4 cell lines was calculated (Supplementary Figure S1B ).
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