Development of a new microfluidic platform integrating co-cultures of intestinal and liver cell lines Thibault Bricks a , Patrick Paullier a , Audrey Legendre a , Marie-José Fleury a , Perrine Zeller a , Franck Merlier b , Pauline M. Anton c , Eric Leclerc a,⇑ a CNRS-UMR 7338, Laboratoire de Biomécanique et Bioingénierie, Université de Technologies de Compiègne, France b CNRS FRE 3580, Laboratoire de Génie Enzymatique et Cellulaire, Université de Technologies de Compiègne, France c EGEAL, Institut Polytechnique Lasalle Beauvais, France article info Article history: Received 25 September 2013 Accepted 17 February 2014 Available online 22 March 2014 Keywords: Co-cultures Microfluidic biochips Caco-2 TC7 HepG2 C3A Phenacetin Drug screening abstract We developed a new biological model to mimic the organ–organ interactions between the intestine and the liver. We coupled polycarbonate cell culture inserts and microfluidic biochips in an integrated fluidic platform allowing dynamic co-cultures (called IIDMP for Integrated Insert in a Dynamic Microfluidic Platform). The intestinal compartment was simulated using Caco-2 TC7 cells and the liver one by HepG2 C3A. We showed that Caco-2 TC7 viability, barrier integrity and functionality (assessed by paracellular and active transport), were not altered during co-cultures in the bioreactor in comparison with the con- ventional insert Petri cultures. In parallel, the viability and metabolism of the HepG2 C3A cells were maintained in the microfluidic biochips. Then, as proof of concept, we used the bioreactor to follow the transport of phenacetin through the intestinal barrier and its metabolism into paracetamol by the CYP1A of the HepG2 C3A cells. Our results demonstrated the performance of this bioreactor with cell co-cultures compared to static co-culture controls in which weak biotransformation into paracetamol was detected. Our study illustrated the interest of such a bioreactor combining the advantages of a cell culture barrier and of liver microfluidic cultures in a common framework for in vitro studies. Ó 2014 Elsevier Ltd. All rights reserved. 1. Introduction The use of new in vitro methods to understand and investigate the metabolism and toxicity of drugs increased considerably in the last few years (Lau et al., 2004). Indeed, these methods can allow the culture of cells in controlled environments mimicking the in vivo microenvironment (Inamdar and Borenstein, 2011). Then, they can be used to predict the toxicity and metabolism of drugs with a higher reliability than traditional in vitro methods (Bhogal et al., 2005). Their uses should be also linked to a decrease in the number of preclinical tests and the probability of unforeseen tox- icity during the drug discovery steps. In addition, they can be used to generate various types of data (genomics, physiological based kinetics) for integration into complex and relevant in silico complex framework (Blaauboer, 2003). Traditionally, in vitro methods are based on static cell cultures in plates or Petri dishes. These methods provide a simple, accessi- ble interface for testing the metabolism and toxicity of molecules. However, they are not able to mimic the complexity of in vivo situations such as organ complexity or organ–organ interactions. In this context, numerous new in vitro methods have been devel- oped to open the possibility to cultivate cells in three dimensional environments and/or dynamic conditions under a controlled flow of culture medium (Chang and Hughes-Fulford, 2009; Ghaemmaghami et al., 2012; Inamdar and Borenstein, 2011; Shuler et al., 1996; Sweeney et al., 1995). These types of culture have been associated with better functionality and viability of the cells (Kim et al., 2012; Xu et al., 2012). In addition to these in vitro methods, the use of microfluidic biochips are able to repro- duce several physiological features such as the cellular microenvi- ronment, dynamic conditions found in vivo, the hepatic zonation, 3D cultures (Allen et al., 2005; Inamdar and Borenstein, 2011; Toh et al., 2009; Xu et al., 2012). These biochips can be fabricated with materials that are non-toxic and that have flexible properties, such as Polydimethylsiloxane (PDMS) (Ghaemmaghami et al., 2012; Huh et al., 2011; Inamdar and Borenstein, 2011). Different kinds of tissue cultures have been investigated in the biochips and thus used to reproduce multi-organ interactions (Huh et al., 2011). Each compartment of these biochips can represent the http://dx.doi.org/10.1016/j.tiv.2014.02.005 0887-2333/Ó 2014 Elsevier Ltd. All rights reserved. ⇑ Corresponding author. Tel.: +33 (0)3 44 23 7943. E-mail address: eric.leclerc@utc.fr (E. Leclerc). Toxicology in Vitro 28 (2014) 885–895 Contents lists available at ScienceDirect Toxicology in Vitro journal homepage: www.elsevier.com/locate/toxinvit