The present invention relates to expandable liver organoids, a medium composition for differentiation thereof, and a method for producing liver organoids using the same, and the liver organoids according to the present invention exhibit the characteristics of more mature hepatocytes than 2D differentiated hepatocytes, can be subcultured up to 90 times or more, and exhibit the expandability for maintaining the characteristics of mature hepatocytes even after multiple subcultures, and thus can be usefully utilized for predicting toxicity, regeneration, and inflammatory response, drug screening, and modeling of diseases such as hepatic steatosis.

The present invention describes an efficient and rapid protocol to generate hepatocyte cells from human induced pluripotent stem cells (iPSCs). According to the method, the iPS cells can be differentiated into functional hepatocyte cells in a short time course (less than 15-20 days). The iPSC-derived hepatocyte cells of the invention have a similar gene expression profile to mature hepatocytes. Moreover, the iPSC-derived hepatocyte cells is proved to rescue lethal fulminant hepatic failure in a non-obese diabetic severe combined immunodeficient mouse model. The rapid and efficient differentiation protocol for generation of iPSC-derived hepatocyte cells may offer an alternative option for treatment of liver diseases.

This application relates to non-human primate (NHP) induced pluripotent stem cell (IPSC)-derived hepatocytes, for example, Cynomolgus monkey (Macaca fascicularis) induced pluripotent stem cell-derived hepatocytes, and methods of producing the same. Moreover, this application relates to methods of using NHP IPSC-derived hepatocytes for drug screening, drug safety assessment and in models of infection.

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.

The present application pertains to a sialyloligosaccharide for augmenting the stemness of stem cells and a use thereof and provides a composition for augmenting stemness of stem cells, a method of culturing stem cells in a medium containing a sialyloligosaccharide, a method of augmenting stemness of stem cells, a stem cell with augmented stemness, obtained by the method, and a cell therapy composition including the stem cells as an active ingredient. The composition or the method according to one or more embodiments may suppress the aging of stem cells and may maintain and augment stemness.

Liver metabolism studies are limited by the inability to expand primary hepatocytes in vitro while maintaining their metabolic functions. Human hepatic three dimensional organoids have been established for use in these studies, but hepatic organoids from adult donors had impaired expansion. Methods of achieving expansion of adult donor-derived hepatic organoids (HepAOs) and HepG2 cells (HepGOs) from single cells in organoid cultures using combinations of growth factors and small molecules are described, with assessment of expansion dynamics, gluconeogenic and HNF4α expression, and albumin secretion. The invention discloses conditions including limiting A8301 and incorporating FSK and OSM to allow the expansion of HepAOs from adult donors and HepGOs with gluconeogenic competence. These models increase the repertoire of human hepatic cellular tools available for use in liver metabolic assays.

The present invention relates to a composition for inducing direct conversion from a somatic cell into one or more kinds selected from the group consisting of an induced Hepatic stem cell (iHSC), a hepatocyte, and a cholangiocyte, and a method of direct conversion of a somatic cell into one or more kinds selected from the group consisting of an induced Hepatic stem cell, a hepatocyte, and a cholangiocyte.

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.

This disclosure relates generally to new methods of maintaining the expression of hepatic genes in human hepatocytes and method for maintaining the functional hepatic enzyme activity of primary hepatocytes in culture. The disclosure also encompasses new methods of deriving a population of pure hepatocytes without selecting or sorting the cells from the cultured pluripotent cells.

The present invention is in the field of pluripotent stem cells. In particular the invention relates to a method for (closed system) induction of differentiation of pluripotent stem cells towards a pre-selected cell type, such as, for example, cardiomyocytes or endothelial cells. The method as disclosed herein is particularly useful to upscale the production of cells derived from pluripotent stem cells, in particular (human) cardiomyocytes and/or endothelial cells derived from pluripotent stem cells.