Establishment of a human hepatoma cell line, HCC-T

A continuous human cell line designated HCC-T was established from the hepatocelluar carcinoma (HCC) of a 69-year-old Japanese male patient. Serum hepatitis A antibody, hepatitis B surface, e and core antigens and antibodies to those viral antigens were all negative in this patient. He had no histories of a large alcoholic consumption and blood transfusion. It is thought that the HCC had developed from liver cirrhosis which was a sequela of nonA nonB type chronic hepatitis. HCC-T cells were cultured in tissue culture flasks attaching to the bottom, in RPMI-1640 medium supplemented with 10% fetal calf serum. The population doubling time of HCC-T cell was estimated as 24h in this culture medium. Morphological study based on the light and electron microscopies revealed that HCC-T cells were HCC cells of epithelial-like shape. Intracellular production of alphafetoprotein was demonstrated by direct immumofluorescence technique. The chromosome analysis revealed that HCC-T had a male sex chromosome and a chromosome mode of 64. It was demonstrated that HCC-T cells were able to grow without anchoring dependence in a soft agar medium. When HCC-T cells were subcutaneously transplanted to athymic BALB/c nude mice, tumors developed at sites and minimum transplantable cell number was 2×10⁶. Light microscopic observation showed that the transplanted tumor tissue was similar in morphology to the original HCC tissue.