Phenotypical stability of a human hepatoma cell line, HuH-7, in long-term culture with chemically defined medium.
1984; National Institutes of Health; Volume: 75; Issue: 2 Linguagem: Inglês
Autores
Hidekazu Nakabayashi, Kazuhisa Taketa, Takashi Yamane, Masahiro Miyazaki, Keiko Miyano, Jiro Sato,
Tópico(s)Muscle metabolism and nutrition
ResumoA 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.
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