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Cytotechnology

, Volume 34, Issue 1–2, pp 159–163 | Cite as

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

  • H. T. Kelly
  • E. Hill
  • F. Reid
  • M. H. Grant
  • C. MacDonald
Article

Abstract

Primary rat hepatocytes dedifferentiate rapidly losing theactivities of the drug metabolising enzymes involved in thedetoxification of xenobiotics in the liver. An alternativeapproach to using primary hepatocytes for toxicity testing isthe development of immortalised hepatocyte cell lines via thetransfection of primary hepatocytes with SV40 DNA. In order toassess the suitability of immortalised lines as an alternativeto primary cell cultures we have used RNA arbitrarily primedpolymerase chain reaction to compare gene expression inimmortalised rat hepatocyte cell lines with that in primary rathepatocytes. We have found that differences exist in the RNAtranscripts between fresh and immortalised hepatocyteshighlighting RNA arbitrarily primed polymerase chain reaction asa useful screening method for identifying immortalised lineswhich retain the most `normal' phenotype in relation to theprimary cells from which they originated.

gene expression immortalised hepatocytes RAP-PCR RT-PCR 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • H. T. Kelly
    • 1
  • E. Hill
    • 1
  • F. Reid
    • 1
  • M. H. Grant
    • 2
  • C. MacDonald
    • 3
  1. 1.Biological SciencesUniversity of PaisleyPaisleyUK
  2. 2.Dept. HaematologyGlasgow Royal InfirmaryGlasgowUK
  3. 3.Biological SciencesUniversity of PaisleyPaisleyUK

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