Expression of cathepsin B and aryl hydrocarbon hydroxylase activities, and of apolipoprotein B in human hepatoma cells maintained long-term in a serum-free medium.
We have established the human hepatoma cell line, HepG2, in a defined, serum-free medium. These cells were maintained and studied over a 100-generation period (i.e. 10 serial transfers). Cells maintained in serum-free medium exhibited growth parameters (i.e. saturation density, efficiency of plating, and population doubling time) similar to those obtained with HepG2 cells maintained in serum-supplemented medium. Serum-free cells were also similar to their serum-supplemented counterparts with respect to the expression of cathepsin B activity and the induction of aryl hydrocarbon hydroxylase by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Significantly, HepG2 cells maintained in serum-free conditions also retained the ability to synthesize and secrete proteins, including the liver plasma protein, apo-lipoprotein B. These results indicate that the serum-free medium used in this study supports the long-term growth and maintenance of human hepatoma, HepG2, cells in culture. Inasmuch as these cells retain phenotypes, including differentiated markers previously reported for their serum-supplemented counterparts, they may provide a more reliable, standardized culture system to study the expression, secretion, and regulation of proteins during biological and pathologic processes.
Dufresne
M
M
Department of Biological Sciences, University of Windsor, Ontario, Canada.
Jane
D
D
Theriault
A
A
Adeli
K
K
D001055:Apolipoproteins B / Q000096:biosynthesis*; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis*; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000378:metabolism*; D002401:Cathepsin B / Q000096:biosynthesis*; D002455:Cell Division; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006801:Humans; D011506:Proteins / Q000032:analysis; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000378:metabolism*
D001055:Apolipoproteins B; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D011506:Proteins; D001189:Aryl Hydrocarbon Hydroxylases; D002401:Cathepsin B