DEVELOPMENT OF CONTINUOUS LIVER-CELL CULTURES FROM THE MARINE TELEOST, SPOT (LEIOSTOMUS-XANTHURUS, PISCES, SCIAENIDAE)

This study was designed to identify the optimal conditions for spot (L eiostomus xanthurus) liver cell survival and propagation in vitro. Liv ers were digested by trypsin disaggregation. Liver cells were obtained in yields that ranged from 4 to 64 X 10(4) cells/g of live body weigh t (average 15 X 10(4) cells/g). The isolated liver cells attached to t he substrate within 2-6 h and formed confluent monolayers by the 6-8th day of culture. Intercellular junctions were frequently observed betw een neighboring hepatocytes. The best results were obtained when the c ells were seeded at a density of 1 X 10(6) cells/25 cm(2) flask. RPMI- 1640 and L-15 media supported cellular growth in the presence or absen ce of 5% CO2. Cultured spot liver cells formed a monolayer over a wide range of osmolalities (i.e. 295 to 355 mOsm/kg). Incubation at either 27 degrees C or 21 degrees C best supported the growth of the culture d cells. Cultured spot liver cells were able to synthesize DNA, RNA, a nd protein. Coral tree extract, concanavalin A, and bacterial lipopoly saccharides augmented DNA synthesis in cultured liver cells. Subcultur ing was possible and three continuous cultures (designated as SLW, SLN , and SLF) with the potential to become immortal cell lines were devel oped. Undoubtedly, immortal liver cell lines will be useful in vitro m odels that will allow further investigation of the mechanisms of hepat ic metabolism, tumor formation, and xenobiotic activation.