Establishment of protocols for the differentiation of hepatic cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells could contribute to regenerative cell therapies or drug discovery and development. However, the differentiation efficiency of endoderm-derived cells, such as hepatic cells, from human ES and iPS cells is poor because hepatic cells are differentiated via multiple lineages including endodermal cells, hepatic progenitor cells, and mature hepatocytes. We show here the protocols for efficient hepatic differentiation from human ES and iPS cells by adenovirus vector-mediated gene transfer.