title: Metabolic characterization of primary rat hepatocytes cultivated in parallel microfluidic biochips.
abstract: The functionality of primary rat hepatocytes was assessed in an Integrated Dynamic Cell Cultures in Microsystem (IDCCM) device. We characterized the hepatocytes over 96 h of culture and evaluated the impact of dynamic cell culture on their viability, inducibility, and metabolic activity. Reverse Transcription quantitative Polymerase Chain Reaction (RTqPCR) was performed on selected genes: liver transcription factors (HNF4α and CEBP), nuclear receptors sensitive to xenobiotics (AhR, PXR, CAR, and FXR), cytochromes P450 (CYPs) (1A2, 3A2, 3A23/3A1, 7A1, 2B1, 2C6, 2C, 2D1, 2D2, and 2E1), phase II metabolism enzymes (GSTA2, SULT1A1, and UGT1A6), ABC transporters (ABCB1b and ABCC2), and oxidative stress related enzymes (HMOX1 and NQO1). Microperfused-cultured hepatocytes remained viable and differentiated with in vivo-like phenotype and genotype. In contrast with postadhesion gene levels, the first 48 h of perfusion enhanced the expression of xenosensors and their target CYPs. Furthermore, CYP3A1, CYP2B1, GSTA2, SULT1A1, UGT1A1, ABCB1b, and ABCC2 were upregulated in IDCCM and reached above postextraction levels all along the duration of culture. Metabolic activities were also confirmed with the detection of metabolism rate and induced mRNAs after exposure to several inducers: 3-methylcholanthrene, caffeine, phenacetin, paracetamol,, and midazolam. Finally, this metabolic characterization confirms that IDCCM is able to maintain rat hepatocytes functions to investigate drug metabolism.
authors: CNRS UMR 7338, Laboratoire de Biomécanique et Bio ingénierie, Université de Technologie de Compiègne, France.
authors: Audrey
authors: A
authors: Legendre
authors: Régis
authors: R
authors: Baudoin
authors: Giulia
authors: G
authors: Alberto
authors: Patrick
authors: P
authors: Paullier
authors: Marie
authors: M
authors: Naudot
authors: Thibault
authors: T
authors: Bricks
authors: Jessy
authors: J
authors: Brocheton
authors: Sébastien
authors: S
authors: Jacques
authors: Jérôme
authors: J
authors: Cotton
authors: Eric
authors: E
authors: Leclerc
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002470:Cell Survival; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004867:Equipment Design; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D015262:Xenobiotics / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D004364:Pharmaceutical Preparations; D018160:Receptors, Cytoplasmic and Nuclear; D015262:Xenobiotics; D003577:Cytochrome P-450 Enzyme System
keywords: cytochrome P450; drug delivery system; efflux pumps; hepatic metabolism; hepatocytes; in vitro models; phase II metabolism