A cell line, HuH-28 was established in vitro from a patient with cholangiocellular carcinoma (CCC). This cell line has been in continuous culture over 10 month period with slow growth potential. HuH-28 was composed of spindle-shaped cells as major population besides a small percentage of polygonal-shaped cells. Chromosome number of the cells were distributed near the hypotriploid region on the 3rd passage. HuH-28 cells were not transplantable into nude mice, but secreted some tumor markers including alkaline phosphatase (ALP), gamma glutamyltranspeptidase (GGT), beta 2-microglobulin (BMG), ferritin, elastase-1 and tissue polypeptide antigen (TPA). This HuH-28 cell line will represent a good model for the investigation of carcinogenesis, histogenesis+ and diagnosis of CCC.