Abstract
Pluripotent stem cell (PSC)-derived hepatocyte-like cells (HLCs) have shown great potential as an alternative to primary human hepatocytes (PHHs) for in vitro modeling. Several differentiation protocols have been described to direct PSCs toward the hepatic fate. Here, by leveraging recent knowledge of the signaling pathways involved in liver development, we describe a robust, scalable protocol that allowed us to consistently generate high-quality bipotent human hepatoblasts and HLCs from both embryonic stem cells and induced PSC (iPSCs). Although not yet fully mature, such HLCs were more similar to adult PHHs than were cells obtained with previously described protocols, showing good potential as a physiologically representative alternative to PHHs for in vitro modeling. PSC-derived hepatoblasts effectively generated with this protocol could differentiate into mature hepatocytes and cholangiocytes within syngeneic liver organoids, thus opening the way for representative human 3D in vitro modeling of liver development and pathophysiology.
•We generated human hepatoblasts and hepatocyte-like cells (HLCs) from pluripotent stem cells•Timed action on Wnt/β-catenin and TGFβ pathways improved maturity and yield of HLCs•Hepatoblasts matured into hepatocytes and bile ducts within complex liver organoids•The protocol is robust and showed potential for scalability and drug testing
The authors describe a new robust differentiation protocol to generate human hepatoblasts and hepatocyte-like cells (HLCs) from pluripotent stem cells. By leveraging the latest knowledge about the signaling pathways involved in liver development, they managed to improve maturity and yield of the differentiation, thus obtaining cells that show great potential to model and study human liver development, physiology, and disease both in 2D and 3D.