Comparison of RAP-PCR analysis of gene expression in fresh and immortalised rat hepatocyte cell lines

Abstract

Primary rat hepatocytes dedifferentiate rapidly losing the activities of the drug metabolising enzymes involved in the detoxification of xenobiotics in the liver. An alternative approach to using primary hepatocytes for toxicity testing is the development of immortalised hepatocyte cell lines via the transfection of primary hepatocytes with SV40 DNA. In order to assess the suitability of immortalised lines as an alternative to primary cell cultures we have used RNA arbitrarily primed polymerase chain reaction to compare gene expression in immortalised rat hepatocyte cell lines with that in primary rat hepatocytes. We have found that differences exist in the RNA transcripts between fresh and immortalised hepatocytes highlighting RNA arbitrarily primed polymerase chain reaction as a useful screening method for identifying immortalised lines which retain the most 'normal' phenotype in relation to the primary cells from which they originated.