Measurement of Cytochrome P450 Enzyme Induction and Inhibition in human cells

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Scope of the method

The Method relates to

Human health

The Method is situated in

Basic Research

Type of method

In vitro - Ex vivo

This method makes use of

Human derived cells / tissues / organs

Animal and human derived cells

Specify the type of cells/tissues/organs

parenchymal liver cells, stem cell-derived hepatocyte-like cells

Description

Method keywords

in vitro
hepatic cell line
luminescence
liver enzyme
viability test

Scientific area keywords

Toxicology
Drug metabolism
drug screening
in vitro cell culture

Method description

By the use of monolayer cultures as an in vitro system, the effects of drugs on CYP3A activity can be evaluated. It relies on the use of a luminogenic CYP3A substrate, namely luciferin-6′-pentafluorobenzylether (luciferin- PFBE). Upon biotransformation by CYP3A, luciferin-PFBE is converted into luciferin, which generates light when combined with a luciferin detection reagent. The normalization of the data relies on the cell number and cell viability and is evaluated by another bioluminescence reaction in which the levels of adenosine- 5′-triphosphate (ATP), the basic energy source of living cells, is measured.

Lab equipment