Establishment of two rat hepatoma cell strains produced by a carcinogen initiation, phenobarbital promotion protocol

Two Fischer 344 rat hepatoma cell strains, JM1 and JM2, have been isolated from a primary hepatocellular carcinoma. Primary tumor formation was induced in a two-thirds partially hepatectomized rat by a single low dose (70 mg/kg of diethylnitrosamine followed by chronic phenobarbital administration (0.1 g/100 ml drinking water). The primary tumors were passed three times by subcutaneous implantation of tumor fragments into the inguinal region of syngeneic recipients. The fourth pass was by injection of tumor cells directly into the livers of recipient rats. Several weeks later, the tumor-containing rat livers were subjected to collagenase perfusion. Two cell lines emerged from tissue culture of the cells isolated by perfusion. Each cell line was cloned by serial dilution. Cells JM1 and JM2 were tumorigenic when injected into syngeneic rats. The tumors, which arose from injected cell strains, exhibited several characteristics of hepatocellular carcinoma. Morphology was examined by light and electron microscopy. Histochemical studies of JM1 and JM2 cells grown in vitro and in vivo were done. The levels of tyrosine aminotransferase and three microsomal enzymes of importance to drug and carcinogen metabolism were investigated. To our knowledge, this is the first report of cell strains derived from an initiation promotion protocol in rats.