Phenotypical stability of a human hepatoma cell line, HuH-7, in long-term culture with chemically defined medium

Gan. 1984 Feb;75(2):151-8.


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.

MeSH terms

  • Animals
  • Carcinoma, Hepatocellular / ultrastructure*
  • Colony-Forming Units Assay
  • Culture Media
  • Culture Techniques / methods
  • Humans
  • Immunoelectrophoresis
  • Insulin / pharmacology
  • Liver Neoplasms / ultrastructure*
  • Mice
  • Phenotype
  • Selenium
  • Time Factors
  • alpha-Fetoproteins / analysis


  • Culture Media
  • Insulin
  • alpha-Fetoproteins
  • Selenium