Improvement of HepG2/C3a cell functions in a microfluidic biochip

Abstract

Current developments in tissue engineering and microtechnology fields allow the use of microfluidic biochip as microtools for in vitro investigations. In the present study, we describe the behavior of HepG2/C3a cells cultivated in a poly(dimethylsiloxane) (PDMS) microfluidic biochip coupled to a perfusion system. Cell culture in the microfluidic biochip for 96 h including 72 h of perfusion provoked a 24 h delay in cell growth compared to plate cultures. Inside the microfluidic biochip, few apoptosis, and necrosis were detected along the culture and 3D cell organization was observed. Regarding the hepatic metabolism, glucose and glutamine consumptions as well as albumin synthesis were maintained. A transcriptomic analysis performed at 96 h of culture using Affymetrix GeneChip demonstrated that 1,025 genes with a fold change above 1.8 were statistically differentially expressed in the microfluidic biochip cultures compared to plate cultures. Among those genes, phase I enzymes involved in the xenobiotic's metabolism such as the cytochromes P450 (CYP) 1A1/2, 2B6, 3A4, 3A5, and 3A7 were up-regulated. The CYP1A1/2 up-regulation was associated with the appearance of CYP1A1/2's activity evidenced by using EROD biotransformation assay. Several phase II enzymes such as sulfotransferases (SULT1A1 and SULT1A2), UDP-glucuronyltransferase (UGT1A1, UGT2B7) and phase III transporters (such as MDR1, MRP2) were also up-regulated. In conclusion, microfluidic biochip could and provide an important insight to exploring the xenobiotic's metabolism. Altogether, these results suggest that this kind of biochip could be considered as a new pertinent tool for predicting cell toxicity and clearance of xenobiotics in vitro.

Predictive toxicology using systemic biology and liver microfluidic “on chip” approaches: Application to acetaminophen injury

“…2, Table 2). Our previous work has shown thatnecrosis and apoptosis occurred only to a small extent in the biochips (Prot et al 2011a). …”

“…Bioartificial organs can now take advantage of recent developments in microtechnology to produce systems on a very small scale (Griffith and Naughton 2002;Powers et al, 2002;Sivaraman et al, 2005;Chao et al, 2010;Prot et al, 2011a;Baudoin et al, 2007;Novik et al, 2010;Baudoin et al, 2011). The cellular organization brought about by the micro-topography of these systems and their dynamic microfluidic culture conditionsappear to be key features for reproducing in vivo environments.…”

“…In our previous work, we showed that hepatocytes grown in microfluidic biochip maintain the activity of their main enzymes for xenobiotic metabolism (CYP1A, CYP2B, CYP3A4, several SULT and UGT sub-families and various phase 3 transporters such as MDR1 and MRP2) (Prot et al, 2011a(Prot et al, , 2011b. In a study of the well-known hepatotoxic drug acetaminophen (APAP) in HepG2/C3a cells, we demonstrated that the use of biochips helps reproduce some of its in vivo reported mechanism of toxicity, such as GSH depletion and mitochondrial damage (Prot et al, 2011c).…”

Chip-based liver equivalents for toxicity testing – organotypicalness versus cost-efficient high throughput

“…57 The biochip environment contributed to the upregulation of principal enzymes involved in phase I and II metabolism and phase III transporters. 58 Data…”

Amperometric Screen-Printed Galactose Biosensor for Cell Toxicity Applications

“…Hepatocellular carcinoma cells have been used to screen complex mixtures, in addition to screening individual chemical compounds for potential hepatotoxic effects. Assays using these cells have been used successfully to provide data for determining potential mechanisms of liver toxicity (Kelly and Sussman 2000;Flynn and Ferguson 2008;Li and Chan 2009;Prot 2011). …”