The pharmaceutical industry has an unmet need for predictive human models for drug metabolism and pharmacokinetics, drug-drug interactions, and drug-induced liver injury (DILI) that can better emulate human response to drugs. Current 2D liver models often fail to capture responses seen in the clinic, as the cellular microenvironment does not accurately reflect what is found in vivo. Here we applied a vascularized human Liver-Chip model that contains primary human hepatocytes, sinusoidal endothelial cells, and Kupffer cells, cultured under physiological fluid flow in a spatial configuration that recapitulates their cytoarchitecture in the liver resulting in long-term viability and improved functionality. The Organ-Chip’s fluidic structure allows for easy effluent sampling for the detection of various biomarkers and cytokine release, and the optical clarity of the Organ-Chip allows for various morphological analyses throughout the culture duration. This provides a platform to investigate mechanistic insights of various DILI and demonstrates the value of the human Liver-Chips for predicting human metabolism, safety testing, risk assessment, and drug discovery and development.