Methods for producing hepatocytes from pluripotent human stem cells are disclosed herein. The stem cells are plated on a cell culture substrate comprising two laminins. The stem cells are then exposed to different cell culture mediums to induce differentiation. The resulting hepatocytes have higher metabolic capacity compared to hepatocytes cultured on different substrates.

The present invention provides methods comprising both genetic and chemical means for the production of hepatocytes from a variety of cell sources, particularly pluripotent stem cells.

Compositions and methods are described herein for inducing reprogramming of non-pluripotent cells across lineage and differentiation boundaries to generate endodermal progenitor cells and hepatocytes. Compositions and methods for expansion of endodermal progenitor cells without loss of phenotype are also described herein.

Provided are: an agent for organoid formation and proliferation in the absence of an extracellular matrix, the agent including a Hippo signaling pathway inhibitor as an active component; a method for organoid proliferation; an organoid proliferated by the above-described proliferation method; an agent for establishing an organoid in the absence of an extracellular matrix; a method for producing an organoid; an organoid produced by the above-described production method; a regenerative medicine preparation; and a method for screening for an agent that enables culture of an organoid in the absence of an extracellular matrix.

Provided is an isolated population of human pluripotent stem cells comprising at least 50% human pluripotent stem cells characterized by an OCT4+/TRA1-60/TRA1-81/SSEA1+/SSEA4 expression signature, and novel methods of generating and maintaining same in a pluripotent, undifferentiated state a suspension culture devoid of cell clumps. Also provided are novel culture media, cell cultures and methods for culturing pluripotent stem cells in a suspension culture or a two-dimensional culture system while maintaining the cells in a proliferative, pluripotent and undifferentiated state. The novel culture media comprise interleukin 11 (IL11) and Ciliary Neurotrophic Factor (CNTF); bFGF at a concentration of at least 50 ng/ml and an IL6RIL6 chimera; or an animal contaminant-free serum replacement and an IL6RIL6 chimera. Also provided are methods for generating lineage-specific cells from the pluripotent stem cells.

Disclosed are improved methods of making liver organoids, and methods of making population organoid panels which can be used, for example, for genotype-phenotype analysis or screening of test compounds. Also disclosed are methods of assessing the risk/prognosis of fatty acid liver disease in subjects having a SNP variant GCKR-rs1260326, as well as novel methods of treating such subjects.