Preliminary study to characterise the metabolic competence of in vitro cellular systems
The purpose of this report is to describe the experimental procedure used to perform a preliminary study to characterise the metabolic competence of in vitro cellular systems.
As a first study, the metabolic competence was assessed by monitoring the presence of four specific enzymes by using selected probe substrates. Each probe substrate can be metabolised by only one enzyme and therefore the formation over time of metabolites is indicative for the presence of a certain enzyme. The concentrations of each metabolite over-time were measured by using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS).
The method developed to monitor the presence of the four enzymes was then used to compare the metabolic competence between two cellular systems: HepaRG (human liver cells known to be metabolically competent) and AR-CALUX (human osteoblastic osteosarcoma U2-OS cells having low or none metabolic activity).
The results and experience achieved will serve to ultimately support the planned work of Directorate F3 focused on developing an EURL ECVAM recommendation, which will be followed by an OECD Guidance Document, with the objective to characterise and describe in vitro hepatic metabolic clearance methods in order to facilitate their regulatory uptake and use to support chemical risk assessment.
â
ZORZOLI Maria Chiara;
MENDOZA Emilio;
MORATH Siegfried;
COECKE Sandra;
LOSTIA Alfonso;
2016-12-23
Publications Office of the European Union
JRC104214
978-92-79-64650-8,
1831-9424 (online),
EUR 28365 EN,
OP LB-NA-28365-EN-N (online),
https://publications.jrc.ec.europa.eu/repository/handle/JRC104214,
10.2788/154828 (online),
Additional supporting files
| File name | Description | File type | |
|---|
