Primary cultured cells and an established cell line of human hepatocellular carcinomas

Cancer cells obtained from human hepatocellular carcinoma nodules were subjected to primary culture, and a hepatoma cell line was established. The cell clumps obtained by needle puncture were plated directly in plastic tissue culture flasks without any cell dissociation procedures. Cell clusters became attached to flasks in 24 h with an efficiency of about 90%. No fibroblast outgrowth was observed. Primary cultured cells were composed of polygonally shaped epithelial cells with dense cytoplasm and one or more large nuclei. They excreted plasma protein biosynthetic markers of hepatocytes into the culture medium. Plasma protein synthesis of primary cultured hepatoma cells decreased as the age of the primary cultures increased. Cells seeded in September 1980 started to grow continuously after 5 months of cultivation. A new hepatocellular carcinoma cell line (designated as KG55T) was established from these growing cells. KG55T cells have been subcultured for more than 20 passages and form a monolayer of polygonal epithelial cells which pile up after they reach confluence. The cells had a doubling time of 50-60 h and a plating efficiency of 60-65%. Albumin, alpha 1-antitrypsin and alpha 2-macroglobulin syntheses and tyrosine aminotransferase activity were detected. At the 10th passage, KG55T cells were pseudotriploid (mode, 69), and 8q+ and 15q+ translocations were distinctive of this cell line. The morphological characteristics and the capacity for plasma protein synthesis of the primary cultured hepatoma cells and cells of the established hepatoma cell line were compared.