Establishment and Characterization of Pancreatic Cell Lines

BxPC-3, Hs766T, Capan-1, MIA PaCa-2, and THP-1 (human monocytic cell line) cell lines were purchased from the American Type Culture Collection. Paca-44, PT45, HPAF and A8184 cell lines were kindly provided by Dr. Piemonti (San Raffaele Scientific Institute). Cell lines were cultured in IMDM 10% fetal bovine serum (FBS) (Lonza) and, in the case of THP-1, β-mercaptoethanol (50 mM) (Sigma). Primary cultures of tumor cells (PCC#353 and PCC#406) and CAFs were established from tumor samples collected at surgery, as described in [10 (link)]. Briefly, tumor pieces were put in culture in IMDM medium (Lonza) plus 10% FBS and CAFs obtained by outgrowth. Alternatively, to obtain distinct cell populations after few passages tumor cells and CAFs were separated with anti-fibroblast Ab-coated beads (Miltenyi Biotec). Primary tumor cells and CAFs were characterized by western blot (WB) for expression of pan-cytokeratin and α-SMA, respectively, as shown in [10 (link)]. Cell lines were periodically tested for Mycoplasma contamination using the MycoAlert™ Mycoplasma Detection kit (Lonza).