Abstract: With the introduction of the principles of the "3R" and the "Toxicology Testing in the 21st Century: Vision and Strategy", the focus of toxicological evaluation has shifted from in vivo testing to the application of in vitro models, thereby promoting the development of in vitro alternative models and attracting increasing attention from scholars. The liver is an important site for the metabolism of exogenous chemicals, and some exogenous chemicals metabolized by the liver can cause liver damage characterized by inflammation, oxidative stress, and necrosis. Therefore, it is crucial to use hepatotoxicity alternative models to detect the hepatotoxicity of exogenous chemicals. This article focused on the research of the in vitro alternative models of 2D hepatocytes, liver slices, and 3D liver for the evaluation of hepatotoxicity, and also provided a systematic description of the biomarkers of drug-induced liver injury (DILI), diagnostic methods for DILI, and emerging technologies related to DILI.