The appearance or relief of inflammation through drug therapy could differentially affect levels of enzymes involved in metabolism of co-administered drugs with potential pharmacological and toxicological consequences. An in vitro model that mimics liver inflammation may provide better predictive data in preclinical testing. Here, we supplement the HepatoPac platform with primary Kupffer macrophages in order to mimic one component of inflammation. Species-matched Kupffer cells were added to human and rat HepatoPac at multiple ratios (to mimic both the normal and inflamed state of the liver) after stabilization to generate a tri-culture with primary hepatocytes and embryonic fibroblasts (HepatoPac- Kupffer cell co-culture). Recent evidence suggests that interaction between inflammatory stress and certain drugs may precipitate toxic responses. We also assess whether stimulation of HepatoPac-Kupffer cell co-cultures with LPS sensitizes the cultures to trovafloxacin (TVX) toxicity. Rat or human HepatoPac-Kupffer cell co-cultures were treated with increasing concentrations of TVX (+/- LPS) and assessed for changes in hepatic ATP content. In conclusion, rat or human HepatoPac- Kupffer cell co-cultures may be used to predict drug induced liver injury mediated by inflammatory stress.