title: A cocktail of metabolic probes demonstrates the relevance of primary human hepatocyte cultures in a microfluidic biochip for pharmaceutical drug screening.
abstract: In this paper, we compare the biotransformation capacities of cryopreserved primary human hepatocytes cultivated in a liver microfluidic biochip and in plates. The hepatocytes were exposed to the CIME cocktail (Carte d'Identité MEtabolique), a mixture of seven probes (acetaminophen, amodiaquine, caffeine, dextromethorphan, midazolam, omeprazole and tolbutamide) for key enzymes involved in the xenobiotic metabolism and pharmacokinetics. The purpose of the cocktail was to give an overview of the metabolic profile of the hepatocytes due to concomitant exposure and a simultaneous mass spectrometric detection method of the metabolites. The results showed a greater activity for CYP1A2, CYP2C9, CYP2C19 CYP2D6, CYP3A and UGT1A1 after 4 h of incubation in the microfluidic biochip when compared to the plate cultures. Furthermore, the metabolic ratio time-course measured at 1 h, 3 h and 4 h indicated that the enzymatic activity increased when the hepatocytes were cultivated in the microfluidic biochip, in contrast with their response in the plate cultures. These results illustrated the functional relevance of liver culture in the PDMS microfluidic biochip. The original method based on a microfluidic culture coupled with CIME cocktail analysis allowed the maintenance and the evaluation of the metabolic performances of the primary human hepatocytes through a new rapid assay. This metabolic analysis can thus become the reference situation when parallel studies of drug metabolism and toxicities are planned with functional hepatocytes in biochips.
authors.affiliation: CNRS UMR 6600, Laboratoire de Biomécanique et Bioingénierie, Université de Technologie de Compiègne, France.
authors.forename: Jean-Matthieu
authors.initials: JM
authors.lastname: Prot
authors.forename: Orianne
authors.initials: O
authors.lastname: Videau
authors.forename: Céline
authors.initials: C
authors.lastname: Brochot
authors.forename: Cécile
authors.initials: C
authors.lastname: Legallais
authors.forename: Henri
authors.initials: H
authors.lastname: Bénech
authors.forename: Eric
authors.initials: E
authors.lastname: Leclerc
mesh_terms: D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D008658:Inactivation, Metabolic; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism*; D012015:Reference Standards; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D004364:Pharmaceutical Preparations