Transfection of hepatic genes into adult rat hepoatocytes in primary culture and their tissue‐specific expression

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Abstract
We describe in this paper a method for studying transient gene expression in a primary culture of adult rat hepatocytes. After isolation by collagenase perfusion, hepatocytes in a monolayer were transfected with foreign DNA by the calcium phosphate precipitation technique during the first 24 hours after plating. When they were transfected with a plasmid containing the gene for chloramphenicol acetyltransferase driven by the early promoter of simian virus 40, hepatocytes reproducibly expressed high levels of chloramphenicol acetyltransferase (CAT); this transient expression was much higher than that obtained with the rat hepatoma cell line H4II. Different medium conditions have been tested: an optimal level of CAT activity can be obtained using a serum‐free, hormonally defined medium. Using these techniques, we have investigated the expression of liver‐specific genes transferred into hepatocytes. We show that the lv‐pyruvate kinase promoter is active in these hepatocytes while it is silent in fibroblasts.Moreover, the use of serum‐free medium may allow investigation of the role of hormones and nutrients in cells which respond normally to these effectors.