Drug-induced hepatotoxicity is a major contributor to the high attrition rates of drug candidates and, for the post-launch withdrawals of drugs in the market. The assessment of hepatotoxicity remains difficult because of the challenges associated with in vivo models like poor correlation, ethical issues and high cost. Most of the current models are focused on hepatocytes alone, lacking the contribution of non-parenchymal cells (NPCs) like Kupffer cells, sinusoidal endothelial cells and stellate cells. Therefore, in vitro human cell-based toxicological models that incorporate NPCs alongside hepatocytes hold the power to enable more realistic cell interactions and cumulative drug response. In this study, we have optimized the best co-culture condition by screening different cell ratio of human liver cancer cell line (HepG2) and human umbilical vein endothelial cells (HUVECs) (1:4,1:1,4:1, respectively) with various cell culture media (EndoGRO-VEGF, EMEM, culture media mixes (1:4,1:1,4:1, respectively), in a Corning® Costar® Ultra-Low attachment 96-well plate to allow the formation of a 3D co-culture cell spheroid. From this screening, we could compare different combinations of cell co-culture ratio and co-culture media, which generated compact co-culture spheroids with an improved hepatic response, as evaluated by the amount of albumin secreted. This hepatic tissue construct was further used as an in vitro model to predict the drug-induced hepatotoxicity of three model drugs (acetaminophen, rifampicin and, ketoconazole). In summary, we have successfully fabricated a highly functional hepatic tissue that has the potential to be used as an in vitro toxicology model as well as used for cell therapy. Authors acknowledge the financial support from Portuguese Foundation for Science and Technology R&D grant POCI-01-0145-FEDER-016715 (PTDC/BBB-ECT/4317/2014) for the project MicroLiver and European Research Council grant agreement ERC-2012-ADG 20120216-321266 for the project ComplexiTE.