We have recently established a cholangiocellular carcinoma (CCC) cell line, designated KMC-1, from a nude mouse subcutaneous tumor which developed after inoculation of a surgically resected peripheral type CCC from a 62-year-old Japanese male patient. KMC-1 cells grew over a 26-month period and passaged 57 times. These cells retained the morphologic characteristics of both the original tumor and the subcutaneous tumor in the nude mouse, which mainly consisted of irregular tubules and invaded surrounding interstitial tissue in part with an indurate pattern. KMC-1 cells grew in a monolayer pavement-like cell arrangement with tubular formation in part. Some cells and/or glands had a mucin-like substance inside. The doubling time of KMC-1 cells growing in serum-containing medium was 54 h at passage 31. Cell growth in serum-free medium was slow but steady. The number of chromosomes was distributed in range from 73 to 83 with modes of 76 and 78. KMC-1 cells secreted some tumor markers such as DUPAN-2, CA125, TPA, hCG, CA19-9 and ferritin, however, the secretion of DUPAN-2, and CA19-9 and ferritin were only detectable in serum-containing and serum-free medium, respectively. These findings suggest that KMC-1 cells will provide a variety of experimental models for research on CCC and the mechanisms of tumor marker secretion.