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Cytotechnology

, Volume 17, Issue 1, pp 45–52 | Cite as

A specific microassay for evaluating hepatic LDH activity in co-cultures of hepatocytes with other cells

  • M. Teresa Donato
  • José V. Castell
  • M. José Gómez-Lechón
Article
  • 46 Downloads

Abstract

This study describes the development of a simple, rapid and reproducible microassay for determining the intracellular LDH activity of rat hepatocytes present in a co-culture system with other cells. The procedure involves treatment of cellular homogenates with an anti-LDH antiserum that specifically inhibits the LDH activity of rat hepatocytes. The assay is performed in 96-well plates and LDH activity can be measured directly in the same wells using a colorimetric method. The difference in LDH activity values measured before and after antiserum incubation reflects the LDH content of the hepatocytes in the sample. The advantages of this method are the small number of cells required, a reduction in sample handling and the possibility of differentiating LDH activity in hepatic and non-hepatic cells. The possible applications of this technique as a parameter for biochemical data and as a test for cytotoxicity studies in co-cultures are also discussed.

Key words

Co-cultures cytotoxicity test hepatocytes LDH activity microassay 

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Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • M. Teresa Donato
    • 1
  • José V. Castell
    • 1
  • M. José Gómez-Lechón
    • 1
  1. 1.Unidad de Hepatología Experimental, Centro de InvestigaciónHospital Universitario La FeValenciaSpain

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