A Long-Term Culture of Human Hepatocytes Which Show a High Growth Potential and Express Their Differentiated Phenotypes

The present study succeeded for the first time in cultivating for more than 2 months human normal hepatocytes which showed a high growth potential and expressed their differentiated phenotypes. Constituents of culture medium were critical for this culture, and the medium optimized for their growth c...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1999-03, Vol.256 (1), p.184-191
Main Authors: Hino, Hiroshi, Tateno, Chise, Sato, Hajime, Yamasaki, Chihiro, Katayama, Shigeru, Kohashi, Toshihiko, Aratani, Akio, Asahara, Toshimasa, Dohi, Kiyohiko, Yoshizato, Katsutoshi
Format: Article
Language:English
Subjects:
3T3 Cells
Adult
Aged
Albumins - analysis
Albumins - secretion
Animals
Ascorbic Acid - analogs & derivatives
Ascorbic Acid - pharmacology
Biomarkers - analysis
Blood Proteins - pharmacology
Cell Culture Techniques - methods
Cell Differentiation - drug effects
Cell Division - drug effects
Cell Size - drug effects
Cell Survival - drug effects
Cells, Cultured
Culture Media, Conditioned
Dimethyl Sulfoxide - pharmacology
DNA - biosynthesis
Epidermal Growth Factor - pharmacology
Female
Galactose - metabolism
Humans
Lidocaine - metabolism
Liver - cytology
Liver - drug effects
Liver - metabolism
Liver - secretion
Male
Mice
Middle Aged
Niacinamide - pharmacology
Time Factors
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Summary:The present study succeeded for the first time in cultivating for more than 2 months human normal hepatocytes which showed a high growth potential and expressed their differentiated phenotypes. Constituents of culture medium were critical for this culture, and the medium optimized for their growth contained fresh human serum, fetal bovine serum, Swiss 3T3-cell conditioned medium, L-ascorbic acid 2-phosphate, epidermal growth factor, nicotinamide, and dimethyl sulfoxide. Hepatocytes steadily replicated and formed colonies which continued to increase in size up to around 35 days. The number of hepatocytes in the most replicative colonies increased 17-fold during 31 days. Cells in colonies expressed normal differentiated hepatocytic phenotypes for as long as 35 days. These hepatocytes retained normal liver functions at least for 70 days such as to secrete albumin, and to metabolize lidocaine and D-galactose.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1999.0288