Microfluidic devices for modeling three-dimensional tissue structures and methods for making and using the same are described herein.

The present invention relates to nanoparticles for the targeted delivery of antigen to liver cells, in particular, liver sinusoidal endothelial cells (LSEC) and/or Kupffer cells, and for the in vivo generation of regulatory T cells, notably CD4+CD25+FOXP3+ regulatory T cells (Treg). The invention provides pharmaceutical compositions and methods for the prevention and treatment of autoimmune diseases, allergies or other chronic inflammatory conditions, and for generation of regulatory T cells. The nanoparticles used in the invention comprise a) a micelle comprising an amphiphilic polymer rendering the nanoparticle water-soluble, and b) a peptide comprising at least one T cell epitope associated with the outside of the micelle. The micelle may or may not comprise a solid hydrophobic core.

The present disclosure provides co-cultures of a population of hepatocytes, at least one non-parenchymal cell population, and a population of hepatic stellate cells in vitro, methods of preparing the co-cultures, and methods of using the co-cultures for high throughput screening and evaluation of drug candidates. The hepatocytes co-culture system provides an in vitro model in which both cell viability and phenotype are maintained for extended periods relative to conventional monoculture.

The present invention pertains to hepatocytes, liver progenitor cells, cholangiocytes, liver sinusoidal endothelial progenitor cells, liver sinusoidal endothelial cells, hepatic stellate progenitor cells, hepatic stellate cells, and liver cellular tissue models, as well as to methods for preparing these cells. The present invention also pertains to a cell fraction comprising liver progenitor cells, liver sinusoidal endothelial progenitor cells, or hepatic stellate progenitor cells. The present invention also pertains to a pharmaceutical composition or kit comprising the above-mentioned cells, a liver cellular tissue model, or a cell fraction. The present invention also pertains to: a method for screening liver disease treatment agents; a method for evaluating the hepatotoxicity of drugs, hepatocytes for infectious disease models, and a method for preparing the same; infectious disease model tissues and a method for preparing the same; as well as a method for screening infectious liver disease treatment agents.

The present invention provides a simple and robust human liver cell-based system in which persistent hepatitis C infection, persistent hepatitis B infection or ethanol exposure induces a clinical Prognostic Liver Signature (PLS) high-risk gene signature. The cellular model system for hepatocellular carcinoma (HCC)/cirrhosis development and progression may be used in the screening of compounds useful in the treatment and/or prevention of cirrhosis and/or HCC as well as in the identification biomarkers for the prediction of liver disease (especially cirrhosis) progression and HCC. The present invention also relates to specific compounds that have been identified, using such screening methods, as useful in the treatment and/or the prevention of HCC/cirrhosis.

HH culture media and methods are provided to allow for recovery from injury or stress associated with cell isolation procedures and freeze/thaw cycle(s) by the use of a DMSO-supplemented medium, i.e., Phase 1 (recovery-phase). Moreover, HH culture media and methods are provided to allow for support of hepatic functionality of cultured HH at a comparable level to that of the human liver by the use of a DMSO, DMSO2, or TMSO-supplemented medium, i.e., Phase 2 (maintenance-application phase). These HH are suitable for the in vitro study of liver biology, diseases drug metabolism and pharmacokinetics. Furthermore, the culture supernatant of HH during Phase 2, namely, conditioned culture medium of human hepatocytes (CMHH), is provided, which facilitates iHeps maturation and supports the cell fate maintenance of hepatocytes. Methods of preparing CMHH are also provided, including culturing humanized liver chimeric animal derived-human hepatocytes and/or primary human hepatocytes in a medium to be collected.

Provided is a method of preparing a three-dimensional cell spheroid, the method including forming the cell spheroid by co-culturing adipose-derived stem cells or mesenchymal stem cells with hepatocytes. According to the cell spheroid prepared by the method, the secretome secreted by the adipose-derived stem cells affects hepatocyte maturation, and therefore, hepatic functions of the finally formed three-dimensional cell spheroid, i.e., organoid, may be enhanced. Further, a composition including a culture medium of the adipose-derived stem cells may prevent or treat liver diseases including hepatitis, hepatotoxicity, cholestasis, fatty liver, etc., and may enhance hepatic functions.

Microfluidic devices for modeling three-dimensional tissue structures and methods for making and using the same are described herein.

Certain relatively small cells present in the periphery blood of mammals can be activated to form pluripotent stem cell populations. These small cells are generally less than five micrometers in diameter and are CD45-positive, and are referred to herein as CD45.sup.+ cells or dormant tiny cells. Accordingly, provided are cell populations and compositions with enriched dormant tiny cells from blood samples and methods and compositions for activating these dormant tiny cells. Upon differentiation, the activated stem cells can be used for various therapeutic purposes.

A method of culturing three-dimensional cellular tissues, including mixing cells with a cationic substance, an extracellular matrix component and a polyelectrolyte to obtain a mixture, collecting cells from the obtained mixture to obtain cell aggregates, seeding the cell aggregates in a culture medium stored in a culture vessel, and culturing the seeded cell aggregates without suspension to thereby obtain a three-dimensional cellular tissue.