The importance of in vitro hepatotoxicity testing during early stages of drug development in the pharmaceutical industry demands effective bioreactor models with optimized conditions. While perfusion bioreactors have been proven to enhance mass transfer and liver specific functions over a long period of culture, the flow-induced shear stress has less desirable effects on the hepatocytes liver-specific functions. In this paper, a two-dimensional human liver hepatocellular carcinoma (HepG2) cell culture flow model, under a specified flow rate of 0.03mL/min, was investigated. Besides computing the distribution of shear stresses acting on the surface of the cell culture, our numerical model also investigated the cell culture metabolic functions such as the oxygen consumption, glucose consumption, glutamine consumption, and ammonia production to provide a fuller analysis of the interaction among the various metabolites within the cell culture. The computed albumin production of our 2D flow model was verified by the experimental HepG2 culture results obtained over 3 days of culture. The results showed good agreement between our experimental data and numerical predictions with corresponding cumulative albumin production of 2.9x10(-5) and 3.0x10(-5)mol/m(3), respectively. The results are of importance in making rational design choices for development of future bioreactors with more complex geometries. Biotechnol. Biotechnol. Bioeng. 2014;111: 885-895. (c) 2013 Wiley Periodicals, Inc.