Establishment of Biliary Tract Cancer Cell Lines

Cell lines were established from pathologically proven primary biliary tract and ampulla of Vater cancer samples of six Korean patients. Of these, two cancer cell lines originated in extrahepatic bile duct cancer, one in intraheatic bile duct cancer, and one in adenocarcinoma of gall bladder, and two in ampulla of Vater cancer. Solid tumours were finely minced with scissors and disassociated into small aggregates by pipetting. Appropriate amounts of finely minced neoplastic-tissue fragments were seeded into 25 cm² flasks. Tumour cells were initially cultured in ACL-4 medium supplemented with 5% heat-inactivated foetal bovine serum (AR5) (Park et al, 1987 (link), 1995 (link)). After establishment, cultures were maintained in RPMI 1640 supplemented with 10% heat- inactivated foetal bovine serum. If stromal-cell growth was noted in initial cultures, differential trypsinisation was used to obtain a pure tumour-cell population. Cultures were maintained in humidified incubators at 37°C in an atmosphere of 5% CO₂ and 95% air. A-431, HeLa, and K-562 cell lines from the American Type Cell Culture (ATCC) and SNU-1 cell line from Korean Cell Line Bank (KCLB) were used for PCR controls. For growth properties and morphology study in vitro, population doubling times and cell viability were determined and cells grown on 75-cm² culture flasks were observed daily by phase-contrast microscopy (Park et al, 1995 (link)).