Pancreatic Cancer Cell Line Characterization

Six human pancreatic cancer cell lines, AsPC-1, BxPC-3, Mia PaCa-2, PANC-1, SU86.86 and T3M4 (kind gifts from Professor Helmut Freiss, Heidelberg University, Germany) were cultured in DMEM or RPMI 1640 media (Hyclone, Thermo Fisher Scientific Inc, Waltham, MA) supplemented with 10% fetal bovine serum (FBS, Hyclone), in a humidified incubator with 5% CO₂ at 37°C. Extracellular matrix (ECM) and AMD3100, the specific inhibitor of CXCR4 [39 (link)], were purchased from Sigma-Aldrich (St. Louis, MO). Recombinant CXCL12, rapamycin and the antibody against CXCL12 were products of R&D Systems. Torin1 [40 (link)] and antibodies for Western blotting of CXCR7, CXCR4 and VEGF were got from Abcam. The antibody for immunoprecipitation of CXCR7 was produced by Thermo. Other antibodies for immunoblotting were all obtained from Cell Signaling Technology (Beverly, MA).