The present invention discloses a method for assessing the capacity of a substance to treat or prevent hepadnavirus infection. A reporter virus carrying genetic information for a first fragment of a recombinase and a reporter cell expressing a second fragment of the recombinase are used. When the reporter virus infects the reporter cell, the two fragments of the recombinase associate and excise a stop cassette that is flanked by two recombination sites and blocks the expression of a reporter gene. Accordingly, the present invention relates to a method of assessing the capacity of a substance to treat or prevent hepadnavirus infection, a hepadnavirus comprising a nucleic acid encoding a first fragment of a recombinase and a mammalian hepatocyte or hepatoma cell comprising a nucleic acid encoding a second fragment of a recombinase and a nucleic acid comprising a stop cassette flanked by two recombination sites fused to a reporter gene.

Identification of urokinase-type plasminogen, matrix metalloproteinase 9, and β-2-microglobulin as novel biomarkers associated with liver fibrosis and uses thereof in diagnosing and staging liver fibrosis.

A 3D tissue culture selected from the group consisting of hydrogel-based 3D tissue culture and cellular self-assembly 3D tissue culture as well as self-assembly 3D tissue culture. Additionally, disclosed is a method of preparing cells for 3D tissue culture, which method comprises the steps of plating the cells on a suitable surface, optionally, checking for their capability to adhere to said surface, discarding the cells which have not adhered to said surface, detaching the adhered cells and transferring them into a 3D tissue culture process.

A cell evaluation method evaluates the quality of a cell population including a plurality of cells. The cell evaluation method comprises: an index calculation step of calculating an index, based on a captured image of the cell population, the index including at least any one of an average distance representing a packing degree of the cells, a spring constant representing a degree of consistency in distances between the cells, and a hexagonal order parameter representing a degree to which an arrangement of the cells resembles a regular hexagon; and an evaluation step of evaluating the cell population, based on the index calculated in the index calculation step.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

The present disclosure provides an in vitro reagent for evaluating xenobiotic metabolism in a cell culture based assay. The in vitro reagent is an admixture of metabolically competent cells and exogenous drug metabolizing enzyme co-factors follow by cryopreservation in the absence of cryopreservation agent so that the cells would be rendered permeable upon thawing due to plasma membrane disruption (while maintaining the integrity of organelles). The permeabilized plasma membranes allow ready diffusion of the exogenous cofactors into the cells to enhance the activities of cellular drug metabolizing enzymes. Addition of a xenobiotic test compound to the thawed in vitro reagent allows metabolism of the test compound by the metabolically competent cells, with metabolites readily diffusible outside the cells due to the permeabilized plasma membranes.

The invention provides a method for constructing a hepatic progenitor cell-like cell bank, including successively performing following processes to human primary hepatocyte cultures from different donor sources: transformation-culture, cryopreservation treatment, proliferation-culture, a first subculture treatment, virus infection, a second subculture treatment, continuous selection-culture and continuous subculture. In the method for constructing a heterogenous immortal hepatic progenitor cell-like cell bank of the present invention, the human primary hepatocyte culture of each of the donor sources is transformation-cultured before the proliferation-culture, which is beneficial in endowing the human primary hepatocyte cultures with good proliferation performance. Once combined with subsequent controlling of culture parameter, the immortal hepatic progenitor cell-like cell lines obtained from different donor sources may have good in vitro proliferation ability. The invention also provides an application of the hepatic progenitor cell-like cell bank and cell lines obtained by the construction method.

The present invention relates to a cell structure including cells containing at least hepatocytes and vascular endothelial cells, and extracellular matrix component, wherein the extracellular matrix components are disposed between the cells, and a liver sinusoidal network is provided between the cells.

The present invention provides a product comprising plated human hepatocytes on a surface and at least some of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, which are attached to the surface. A method of preparing plated human hepatocytes is also provided. The preparation method comprises applying human hepatocytes to a surface in the presence of feeder cells, co-culturing the applied hepatocytes with the feeder cells, and forming one or more hepatocyte clusters by the co-cultured hepatocytes on the feeder cells, which are attached to the surface. The plated hepatocytes may be used for various purposes, including the preparation of a hepatitis B virus (HBV) infected hepatocyte culture model and drug testing.

Disclosed is a spheroid liver organoid comprising hepatic lineage cells such as human hepatocytes, hepatic stellate cells, and liver sinusoidal endothelial cells. Also provided are methods of using spheroid liver organoids for applications related to drug screening and toxicity screening. In particular, spheroid liver organoids are useful for high-throughput screens to identify compounds having efficacy for treating liver disease.