Liver disorders constitute a worldwide increasing problem. Both the study of the mechanisms underlying liver disease and the development of liver disease therapeutics heavily rely on the use of experimental models. Among those, human-based in vitro systems are more and more preferred over laboratory animals because of ethical reasons. Primary human hepatocytes and their cultures are still considered as the gold standard liver-based in vitro models, as they provide a good reflection of the in vivo situation. Nevertheless, these in vitro systems deal with the gradual deterioration of the differentiated morphological and functional phenotype. This can be overcome by following a number of strategies, such as by using spheroid/organoid and sandwich culture configurations. Further improvement in this area in view of enhancing overall translational value of primary human hepatocytes and their cultures warrants interdisciplinary collaboration.