Establishment of pancreatic cancer cell lines with endoscopic ultrasound‐guided biopsy via conditionally reprogrammed cell culture

Abstract

Recent studies have identified the mutational landscape ofpancreaticcancerand suggested tumor-specific subtypes. However, the major hurdle against personalized treatment is the difficulty to obtain sufficientcancertissues from most inoperable cases. We investigated whether patient-derivedconditionallyreprogrammedcells (CRCs) can be constructed using a small piece of tumor tissue usingendoscopicultrasound (EUS)-guided fine needlebiopsy(FNB). Thirty patients withpancreaticsolid mass (mean size, 34.6mm) were enrolled prospectively. Among 22 patients who were diagnosed withpancreaticductal adenocarcinoma, we established patient-derivedpancreaticcancercelllinesfrom eight patients (36.4%). Immunofluorescence colony staining for CRCs showed that the cytoplasm ofcancercells was clearly stained with anti-cytokeratin 19 monoclonal antibody. In the soft agar colony formation assay, CRCs formed colonies compared with the negative control by day 15. In vivo, implanted CRCs showed tumor engraftment and hematoxylin and eosin staining showedpancreaticcancerductal structure. All established CRCs showed a KRAS mutation. In conclusion, we established patient-derivedpancreaticcancercelllineswith a small tumor tissue obtained by EUS-FNB. With in vitro drug sensitivity and genomic studies, established patient-derivedcelllinescan be used in identification of new targets for diagnosis and treatment ofpancreaticcancer.