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Biochem Biophys Res Commun ; 256(1): 184-91, 1999 Mar 05.
Article in English | MEDLINE | ID: mdl-10066444

ABSTRACT

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.


Subject(s)
Cell Culture Techniques/methods , Liver/cytology , 3T3 Cells , Adult , Aged , Albumins/analysis , Albumins/metabolism , Animals , Ascorbic Acid/analogs & derivatives , Ascorbic Acid/pharmacology , Biomarkers/analysis , Blood Proteins/pharmacology , Cell Differentiation/drug effects , Cell Division/drug effects , Cell Size/drug effects , Cell Survival/drug effects , Cells, Cultured , Culture Media, Conditioned , DNA/biosynthesis , Dimethyl Sulfoxide/pharmacology , Epidermal Growth Factor/pharmacology , Female , Galactose/metabolism , Humans , Lidocaine/metabolism , Liver/drug effects , Liver/metabolism , Male , Mice , Middle Aged , Niacinamide/pharmacology , Time Factors
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