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Cytochrome P450 (CYP) enzyme induction in vitro method using the cryopreserved human hepatocytes

Topic : Toxicokinetics

Test Method Number: TM2009-13 (EU)
Short Name of TM: CryoHeps
Year received: 2009
Responsible Organisation: EURL ECVAM - European Union '
Protocol(s)/SOP(s): 193

Method Description

The human cytochrome P450 (CYP) enzyme induction method assesses the potential of test chemicals to induce the activity of three CYP enzymes (CYP1A2, CYP2B6 and CYP3A sub-family) in cryopreserved primary human hepatocytes.
At the molecular level, CYP enzyme induction is a process controlled by a set of nuclear receptors associated with downstream signal transduction pathways.
Experimentally, at the end of cell exposure to the test chemical, a cocktail of three specific CYP enzymatic substrate probes is applied to the cells and CYP enzyme induction is assessed through the measurement of of the specific metabolites formation by CYP1A2, 2B6 and 3A sub-family by analytical liquid chromatography/mass spectrometry (LC/MS) technique.
The CYP enzyme induction in vitro methods (see also TM2009-14) are reliable and relevant methods for the generation of human metabolism data in animal-free testing strategies assessing adverse health effects of...

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