Phenotypical stability of a human hepatoma cell line, HuH-7, in long-term culture with chemically defined medium.
A chemically defined medium containing selenium was further improved, and the improved medium showed excellent ability to support growth and colony formation of a human hepatoma cell line, HuH-7. The improved defined medium (designated as IS-RPMI) contained small amounts of unsaturated fatty acids, inorganic trace elements including selenium, and HEPES buffer. Many of these additions exert only a small individual effect on the growth of cells, but their cumulative effects were substantial. HuH-7 cells replicated continuously with a doubling time of 47-51 hr in IS-RPMI. A conditioned medium from a semiconfluent population increased the colony-forming ability of HuH-7 cells in IS-RPMI. The addition of insulin to the medium gave good colony growth, thus duplicating the effect of the conditioned medium. Although some alterations of morphological and growth characteristics of HuH-7 cells were observed, the differentiated liver functions, as revealed by production of plasma proteins including high levels of alpha 1-fetoprotein, were maintained for a period of more than 3 years and through 70 passages under the above culture conditions.
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000648:ultrastructure*; D003114:Colony-Forming Units Assay; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D006531:HEPES; D006801:Humans; D007122:Immunoelectrophoresis; D007328:Insulin / Q000494:pharmacology; D008113:Liver Neoplasms / Q000648:ultrastructure*; D051379:Mice; D010641:Phenotype; D012643:Selenium; D013997:Time Factors; D000509:alpha-Fetoproteins / Q000032:analysis
D003470:Culture Media; D007328:Insulin; D000509:alpha-Fetoproteins; D012643:Selenium; D006531:HEPES