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

Cytotechnology
M T DonatoM J Gómez-Lechón

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.

References

Mar 1, 1992·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·D Utesch, F Oesch
Nov 1, 1990·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·M T DonatoJ V Castell
Sep 1, 1988·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·F GouletO Morin
Aug 1, 1989·Differentiation; Research in Biological Diversity·W Kuri-Harcuch, T Mendoza-Figueroa
May 4, 1987·Journal of Immunological Methods·A Montoya, J V Castell
Sep 1, 1984·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·J M BegueA Guillouzo
Jan 1, 1991·Toxicology in Vitro : an International Journal Published in Association with BIBRA·M T DonatoJ V Castell

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