SPHEROID CULTURES OF HUMAN HEPATOBLASTOMA CELLS (HUH-6 LINE) AND THEIR APPLICATION FOR CYTOTOXICITY ASSAY OF ALCOHOLS

Spheroid cultures of human hepatoblastoma cells (HuH-6 line) were esta blished by rotating 3 x 10(6) cells/3 ml culture medium in 25-ml Erlen meyer flasks on a gyratory shaker. The size of the spheroids rapidly i ncreased until 4 days of culture, and thereafter their size gradually increased until 8 days of culture. A considerable amount of lactate de hydrogenase (LDH) was detected in the culture medium at 24 h after see ding because of cell damage by subculturing, but thereafter the amount released was small, indicating that the spheroids were in healthy con dition. Albumin production, one of the differentiated functions of hep atocytes, was higher in spheroid cultures than in monolayer cultures. Using this spheroid culture model, the cytotoxic effects of alcohols o n HuH-6 cells were studied by measuring the activity of LDH released i n the medium from damaged cells. The results indicate that the increas ing order of toxicity of the alcohols was as follows: methanol < ethan ol < propanol.