Abstract: Functional hepatocytes have been used in researches on cell transplantation, bio-artificial liver therapy, liver tissue engineering, drug metabolism and toxicity. Primary hepatocytes (PHs), as a source of functional hepatocytes, cannot meet the growing demands in those researches. Functional hepatocytes differentiated in 2D or 3D culture systems from stem cells are considered an alternative cell source. 3D culture systems, mimicking the biological environment in vivo, have recently garnered great attention because they often promote levels of cell differentiation and tissue organization not possible in conventional 2D culture systems. Here, we review the advances in researches of hepatic differentiation from stem cells in different 3D culture systems, focus on the materials used to make the 3D culture scaffolds.