Metabolic characterization of primary rat hepatocytes cultivated in parallel microfluidic biochips. Issue 9 (19th February 2013)

• Record Type: Journal Article
• Authors: Legendre, Audrey; Baudoin, Régis; Alberto, Giulia; Paullier, Patrick; Naudot, Marie; Bricks, Thibault; Brocheton, Jessy; Jacques, Sébastien; Cotton, JérôME; Leclerc, Eric; Nakashima, Emi; Brouwer, Kim; Hammarlund‐Udenaes, Margareta; Terasaki, Tetsuya
• Is Part Of: Journal of pharmaceutical sciences. Volume 102:Issue 9(2013:Sep.)
• Page Start: 3264
• Page End: 3276
• Publication Date: 2013-02-19
• Subjects: Pharmacy -- Periodicals; 615.1
• DOI: 10.1002/jps.23466
• Languages: English
• ISSNs: 0022-3549

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>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 (<italic>HNF4α</italic> and <italic>CEBP</italic>), nuclear receptors sensitive to xenobiotics (<italic>AhR</italic>, <italic>PXR</italic>, <italic>CAR</italic>, and <italic>FXR</italic>), cytochromes P450 (CYPs) (<italic>1A2</italic>, <italic>3A2</italic>, <italic>3A23</italic>/<italic>3A1</italic>, <italic>7A1</italic>, <italic>2B1</italic>, <italic>2C6</italic>, <italic>2C</italic>, <italic>2D1</italic>, <italic>2D2</italic>, and <italic>2E1</italic>), phase II metabolism enzymes (<italic>GSTA2</italic>, <italic>SULT1A1</italic>, and <italic>UGT1A6</italic>), ABC transporters (<italic>ABCB1b</italic> and <italic>ABCC2</italic>), and oxidative stress related enzymes (<italic>HMOX1</italic> and <italic>NQO1</italic>). Microperfused‐cultured hepatocytes remained viable and differentiated with <italic>in vivo</italic>‐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, <italic>CYP3A1</italic>, <italic>CYP2B1</italic>, <italic>GSTA2</italic>, <italic>SULT1A1</italic>, <italic>UGT1A1</italic>, <italic>ABCB1b</italic>, and <italic>ABCC2</italic> 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. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:3264–3276, 2013</p> </abstract>