Establishment and Characterization of Pancreatic Cancer Cell Lines

Human cell lines PANC-1 and HPDE were obtained from American Type Culture Collection (ATCC, Rockville, MD) or as a generous gift from Mokenge Malafa (Moffitt Cancer Center, Tampa, FL), respectively. L3.6pl metastatic variant was derived as previously described [17 (link), 18 (link)]. The selection of L3.6pl^GemRes gemcitabine-resistant pancreatic cancer cells was conducted as previously described [19 (link)]. All human PC cell lines were authenticated within 6 months by short tandem repeat (STR) analysis. All cells were maintained and stimulated in high-nutrition media [Dulbecco’s modified Eagle’s medium/F12 (DMEM/F12) Advanced (Life Technologies, Carlsbad, CA), 10 % fetal bovine serum (FBS) (Atlanta Biologicals, Atlanta, GA), 4 mM GlutaMAX^™ (Life Technologies), 20 ng/mL human epidermal growth factor (Life Technologies), 40 ng/mL dexamethasone (Sigma-Aldrich, St. Louis, MO) and antibiotic antimycotic solution (Sigma)] in 5 % CO₂/95 % air at 37 °C. The creation of L3.6pl^CXCL10 CXCL10-constitutively expressing cells were created by transfection with pCMV6.puromycin vector (Origene, Rockville, MD) containing human CXCL10 cDNA and grown in selective medium containing puromycin (10 μg/mL). Isolated clones were expanded, and CXCL10 secretion was confirmed by ELISA (BD Biosciences, Franklin Lakes, NJ).