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.

A method for producing pancreatic endocrine cells, the method including introducing one or more genes of a GLIS family or one or more gene products thereof and a Neurogenin3 gene or one or more gene products thereof into somatic cells.

A cultured tissue, comprising: glandular cells; glandular cavities formed from the glandular cells; and ducts formed from epithelial cells, wherein the glandular cavities and the ducts are functionally connected ex vivo.

Provided are a method for expanding a hepatocyte in vitro and an application thereof. A culture system is provided for reprogramming a human hepatocyte into a proliferating intermediate-state cell between a mature hepatocyte and a liver progenitor cell. The liver repopulation ability of the system was verified in animals. The method does not require the introduction of an exogenous gene into a hepatocyte, and the expansion of the hepatocyte can be realized by conventional culture. The obtained hepatocyte can be passaged, and can be cultured to maturation to obtain a functional mature human hepatocyte.

A method of constructing a hepatic precursor-like cell line is provided. The deposited number of the hepatic precursor-like cell line is CCTCC NO: C2019120. The construction method includes using primary hepatocytes as seed cells to establish the hepatic precursor-like cell line, which combines with controlling the conditions of passage culture, subculture, virus infection, proliferation culture and confluence culture. Moreover, a hepatocyte-specific transcription factor, FOXA3, is expressed in the immortalized cell line to make the hepatic precursor-like cell line differentiate easily. An application of the hepatic precursor-like cell line in the field of the bioartificial liver is also provided.

The invention relates to a process for the manufacture of a population of human allogeneic liver-derived progenitor cells (HALPC). The process comprises the use of a xeno- and serum-free culture medium comprising purified native or recombinant human serum albumin.

The invention relates to a method for inducing human cholangiocyte differentiation of progenitor cells called hepatoblasts. More specifically, the invention relates to a method for differentiating hepatoblasts to cholangiocytes by culturing said hepatoblasts with a particular medium having interleukin-6 (IL-6) activity. The differentiation method can specifically induce cholangiocyte dilferentiation from hepatoblasts, and the human cholangiocytes differentiated according to the invention may be useful for drug discovery for treatment of cholangiopathies and bioengineered livers.

The invention relates to a method for inducing human cholangiocyte differentiation of progenitor cells called hepatoblasts. More specifically, the invention relates to a method for differentiating hepatoblasts to cholangiocytes by culturing said hepatoblasts with a particular medium having interleukin-6 (IL-6) activity. The differentiation method can specifically induce cholangiocyte differentiation from hepatoblasts, and the human cholangiocytes differentiated according to the invention may be useful for drug discovery for treatment of cholangiopathies and bioengineered livers.

The disclosure provides materials and methods useful in forming at least one bile duct or treating cholestatic disease or injury by transdifferentiating hepatocytes to cholangiocytes by delivery of an effective amount of an expressible Transforming Growth Factor β Type I Receptor (TGFBR1), Transforming Growth Factor β Type II Receptor (TGFBR2), SMAD3, SMAD1, SMAD2, SMAD5 or SMAD8/9, in either in vivo or in vitro environments. Another aspect provides a method of forming at least one bile duct or treating a cholestatic disease or injury by delivering an effective amount of JAG1, JAG2, DLL1, DLL3, DLL4, NOTCH1, NOTCH2, NOTCH3, NOTCH4 or the respective NOTCH intracellular domains either in vivo or in vitro. Also provided are methods for correcting mutant alleles of genes in the TGFβ and/or Notch pathways, e.g., JAG1 or NOTCH2, using ZFNs, TALENs, CRISPR or any other genome editing technique. Additionally, methods are provided for inducing increased expression of a normal, or wild-type, allele of a TGFβ or Notch pathway gene such as TGFBR1 or JAG1 using CRISPRa technology. Yet another aspect is drawn to a method of forming at least one bile duct or treating a cholestatic disease or injury by delivering an effective amount of a wild-type hepatocyte or a hepatocyte that has not been engineered to overexpress a gene product.