Part B: Directed Differentiation of Human Embryonic Stem Cells into Hepatic Cells

Hepatocytes comprise most of the liver and are mainly responsible for metabolizing dietary molecules, detoxifying compounds and storing glycogen. During the embryonic development, hepatocytes progressively modify their gene expression profile according to their role in each stage of the development. Human embryonic stem (ES) cells provide a major tool in understanding liver development, and may also serve as a source of hepatic cells for cellular therapy. In this chapter we review recent research in the field of hepatic differentiation from human ES cells. We discuss methodologies for differentiation of human ES cells towards hepatic cells using various protocols, and we detail our protocol for deriving hepatic-like cells using genetic manipulations. The chapter includes the protocols for creating hES cell lines carrying a reporter gene, differentiation of human ES cells by embryoid bodies (EBs) formation and sorting of the hepatic-like cells using fluorescence activated cell sorting (FACS).