Microfluidic biochip for the perifusion of precision‐cut rat liver slices for metabolism and toxicology studies

Early detection of kinetic, metabolic, and toxicity (ADME‐Tox) profiles for new drug candidates is of crucial importance during drug development. This article describes a novel in vitro system for the incubation of precision‐cut liver slices (PCLS) under flow conditions, based on a poly(dimethylsiloxane) (PDMS) device containing 25‐µL microchambers for integration of the slices. The microdevice is coupled to a perifusion system, which enables a constant delivery of nutrients and oxygen and a continuous removal of waste products. Both a highly controlled incubation environment and high metabolite detection sensitivity could be achieved using microfluidics. Liver slices were viable for at least 24 h in the microdevice. The compound, 7‐ethoxycoumarin (7‐EC), was chosen to test metabolism, since its metabolism includes both phase I and phase II metabolism and when tested in the conventional well plate system, correlates well with the in vivo situation (De Kanter et al. 2004. Xenobiotica 34(3): 229–241.). The metabolic rate of 7‐EC was found to be 214 ± 5 pmol/min/mg protein in the microdevice, comparable to well plates, and was constant over time for at least 3 h. This perifusion system better mimics the in vivo situation, and has the potential to significantly contribute to drug metabolism and toxicology studies of novel chemical entities. Biotechnol. Bioeng. 2010;105: 184–194. © 2009 Wiley Periodicals, Inc.

[1]  A. J. Gandolfi,et al.  Dynamic Organ Culture is Superior to Multiwell Plate Culture for Maintaining Precision-Cut Tissue Slices: Optimization of Tissue Slice Culture, Part 1 , 1995 .

[2]  J. Walsh,et al.  An improved HPLC assay for the assessment of liver slice metabolic viability using 7-ethoxycoumarin. , 1995, Drug metabolism and disposition: the biological fate of chemicals.

[3]  R. de Kanter,et al.  EMPIRICAL VALIDATION OF A RAT IN VITRO ORGAN SLICE MODEL AS A TOOL FOR IN VIVO CLEARANCE PREDICTION , 2006, Drug Metabolism and Disposition.

[4]  P. Olinga,et al.  Precision-cut tissue slices as a tool to predict metabolism of novel drugs , 2007, Expert opinion on drug metabolism & toxicology.

[5]  F. Oesch,et al.  New Hepatocyte In Vitro Systems for Drug Metabolism: Metabolic Capacity and Recommendations for Application in Basic Research and Drug Development, Standard Operation Procedures , 2003, Drug metabolism reviews.

[6]  H. Langenhove,et al.  Investigation of the permeability and selectivity of gases and volatile organic compounds for polydimethylsiloxane membranes , 2003 .

[7]  Teruo Fujii,et al.  Enhanced maintenance and functions of rat hepatocytes induced by combination of on-site oxygenation and coculture with fibroblasts. , 2008, Journal of biotechnology.

[8]  B. J. Kane,et al.  Liver-specific functional studies in a microfluidic array of primary mammalian hepatocytes. , 2006, Analytical chemistry.

[9]  Shuichi Takayama,et al.  Leakage-free bonding of porous membranes into layered microfluidic array systems. , 2007, Analytical chemistry.

[10]  Shinji Sugiura,et al.  Pressure‐driven perfusion culture microchamber array for a parallel drug cytotoxicity assay , 2008, Biotechnology and bioengineering.

[11]  M. Clemens,et al.  Hepatocyte and kupffer cells co-cultured on micropatterned surfaces to optimize hepatocyte function. , 2006, Tissue engineering.

[12]  J. Taylor,et al.  Alterations in albumin secretion and total protein synthesis in livers of thyroidectomized rats. , 1981, The Biochemical journal.

[13]  I. Kola,et al.  Can the pharmaceutical industry reduce attrition rates? , 2004, Nature Reviews Drug Discovery.

[14]  Ivan Rusyn,et al.  Role of the Kupffer cell in mediating hepatic toxicity and carcinogenesis. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[15]  L. Griffith,et al.  A microfabricated array bioreactor for perfused 3D liver culture. , 2002, Biotechnology and bioengineering.

[16]  E. Verpoorte,et al.  Stabilization of two-phase octanol/water flows inside poly(dimethylsiloxane) microchannels using polymer coatings , 2006, Analytical and bioanalytical chemistry.

[17]  Luke P. Lee,et al.  An artificial liver sinusoid with a microfluidic endothelial-like barrier for primary hepatocyte culture. , 2007, Biotechnology and bioengineering.

[18]  P. Olinga,et al.  Liver slices as a model to study fibrogenesis and test the effects of anti-fibrotic drugs on fibrogenic cells in human liver. , 2008, Toxicology in Vitro.

[19]  M. López-Garcı́a.  Endogenous nitric oxide is responsible for the early loss of P450 in cultured rat hepatocytes , 1998, FEBS letters.

[20]  P. Balladur,et al.  Morphological and Biochemical Integrity of Human Liver Slices in Long-Term Culture: Effects of Oxygen Tension , 2004, Cell Biology and Toxicology.

[21]  John Greenman,et al.  Development of a microfluidic device for the maintenance and interrogation of viable tissue biopsies. , 2008, Lab on a chip.

[22]  Hanry Yu,et al.  A practical guide to microfluidic perfusion culture of adherent mammalian cells. , 2007, Lab on a chip.

[23]  Philip Britz-McKibbin,et al.  On-line preconcentration strategies for trace analysis of metabolites by capillary electrophoresis. , 2003, Journal of chromatography. A.

[24]  Jing Zhang,et al.  Novel intra-tissue perfusion system for culturing thick liver tissue. , 2007, Tissue engineering.

[25]  D. Beebe,et al.  Cell culture models in microfluidic systems. , 2008, Annual review of analytical chemistry.

[26]  G. Whitesides,et al.  Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane). , 1998, Analytical chemistry.

[27]  Laurent Griscom,et al.  In situ micropatterning technique by cell crushing for co-cultures inside microfluidic biochips , 2008, Biomedical microdevices.

[28]  R. de Kanter,et al.  Prediction of whole-body metabolic clearance of drugs through the combined use of slices from rat liver, lung, kidney, small intestine and colon , 2004, Xenobiotica; the fate of foreign compounds in biological systems.

[29]  P. Olinga,et al.  Drug-metabolizing activity of human and rat liver, lung, kidney and intestine slices , 2002, Xenobiotica; the fate of foreign compounds in biological systems.

[30]  Hanry Yu,et al.  Perfusion culture improves the maintenance of cultured liver tissue slices. , 2007, Tissue engineering.

[31]  Aaron Sin,et al.  Development of a Microscale Cell Culture Analog To Probe Naphthalene Toxicity , 2008, Biotechnology progress.

[32]  S. Ostrovidov,et al.  Membrane-Based PDMS Microbioreactor for Perfused 3D Primary Rat Hepatocyte Cultures , 2004, Biomedical microdevices.

[33]  P. Olinga,et al.  Comparison of five incubation systems for rat liver slices using functional and viability parameters. , 1997, Journal of pharmacological and toxicological methods.

[34]  P. H. Bach,et al.  Optimizing preincubation conditions for precision-cut rat kidney and liver tissue slices: effect of culture media and antioxidants. , 1998, Toxicology in vitro : an international journal published in association with BIBRA.