The liver is a common target of toxic effect of a number of xenobiotics, which is in particular a result of its central role in intermediary and energetic metabolism and in biotransformation processes. Ethical, economic, legislative, research and other reasons do not allow testing all of newly-synthesized compounds in in vivo conditions. Hence new methods and approaches for hepatotoxicity testing in vitro have been developing. The most important systems for study of toxicity and metabolic activity in vitro are isolated perfused liver, liver slices, isolated liver cells in suspensions or in primary cultures including co-culture methods and special 3D techniques, various subcellular fractions and stabilised cell lines. These models can be used for cytotoxicity and genotoxicity screening, evaluation of potential hepatoprotective capacity of different compounds, study of toxic injury and characterization of hepatotoxicity mechanisms. Currently there is no an ideal in vitro liver model system for testing of hepatotoxic substances in vitro, nevertheless use of these model systems reduces economic costs and ethic and legislative problems. Model systems in vitro afford opportunity to study in detail mechanisms of hepatotoxicity in comparison with in vivo conditions. Definition of their actual advantages and disadvantages allows choosing a suitable model system for study of particular problem. We cannot imagine current research of liver toxicity without using these model sytems.