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 |
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Authors: | Michael Dufresne Derek Jane Andre Theriault Khosrow Adeli |
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Affiliation: | (1) Department of Biological Sciences, University of Windsor, N9B 3P4 Windsor, Ontario, Canada;(2) Department of Chemistry and Biochemistry, University of Windsor, N9B 3P4 Windsor, Ontario, Canada |
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Abstract: | Summary 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-tetra-chlorodibenzo-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. |
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Keywords: | HepG2 serum-free medium AHH activity cathepsin B apolipoprotein B |
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