α smooth muscle actin

Patient-derived PC cell lines were isolated from patient-derived xenografts as previously described (16 (link),17 (link)). Epithelial cell purity was confirmed by expression of HLA ABC, EpCam and cytokeratins 18 and 19 (Biolegend, San Diego, CA). Patient-derived tumor associated stroma (TAS) cell lines were generated as previously described (15 (link)). Cell purity was confirmed by uniform expression of α-smooth muscle actin (R&D Systems, Minneapolis, MN) by flow cytometry and immunocytochemistry. Both PC and TAS cell lines were maintained and stimulated in growth medium [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), Primocin™ (Invivogen, San Diego, CA) and antibiotic antimycotic solution (Sigma)] in 5% CO2/95% air at 37°C.

Patient-derived PPCE were isolated from patient-derived xenografts [20 (link)]. Epithelial cell purity was confirmed by expression of HLA-ABC and cytokeratins 18 and 19 (Bio-legend, San Diego, CA) by flow cytometry and immunofluorescent microscopy. Patient-derived TAS outgrowths were generated from direct culture of gross human pancreatic adenocarcinoma surgical specimens as previously validated [21 (link)]. Cell purity was confirmed by high expression levels of α-smooth muscle actin (R&D Systems, Minneapolis, MN) by flow cytometry and immunocytochemistry.