A method of treating liver-based inborn, metabolic deficiencies is disclosed by treatment of an individual, such as a patient suffering from liver-based inborn, metabolic deficiencies, with human progenitor or stem cells, a cell population or their progeny. The cells used in the treatment have the following characteristics. They are positive for vimentin, α-smooth muscle actin (ASMA), and for at least one mesenchymal marker such as CD90, CD29, CD73, and CD44. They are positive for at least one hepatocyte marker such as albumin, alpha-fetoprotein, alpha-1 antitrypsin, HNF-4 and MRP2 transporter. They express at least one hepatocyte-like property or function such as G6P, CYP1B1, CYP3A4, TDO, TAT, GS, GGT, CK8, and EAAT2. They are negative for at least one marker such as cytokeratin-19, CD45, CD34, CD49f, CD133, HLA-DR, and CD117. They have mesenchymal-like morphology. They originate from human adult liver cells.

Novel adult liver progenitor cells (called H2Stem Cells) have been have been characterized on the basis of a series of biological activities and markers. Methods for producing H2Stem Cells allow providing such cells in the form of adherent cells and three-dimensional cell clusters in suspension that can be differentiated into cells having strong liver-specific activities and/or that can be used for treating liver diseases or for evaluating the efficacy, the metabolism, and/or toxicity of a compound.

A method for isolating and processing bone marrow derived stem cells, including the steps of: (a) collecting a biological sample containing adherent bone marrow stem cells in a receptacle with interior walls coated with a cell-adherent substrate; (b) incubating the bone marrow cells on the adherent substrate so that a layer of adherent bone marrow stem cells adheres to the substrate; (c) washing any non-adherent cells from the substrate; and (d) collecting the bone marrow stem cell layer. Isolation kits and use of bone marrow cells harvested for cell therapies are also described.

The technology described herein is directed to methods of cryopreservation, e.g., cryopreservation in a microfluidics format and methods of utilizing cells preserved by such methods.

The invention relates to a liver organoid, uses thereof and method for obtaining them.

There is disclosed a preparation of extracellular vesicles derived from human liver stem cells, preferably from a non-oval human progenitor cell line that expresses hepatic cell markers, which is capable of reducing the senescence of a population of senescent cells, as measured in a SA-β-galactosidase-based cellular senescence assay. Also disclosed are therapeutic applications of the preparation of extracellular vesicles derived from human liver stem cells according to the invention. Therapeutic applications include the reduction of cellular senescence, e.g. in an vitro or ex vivo method, as well as the therapeutic treatment of diseases and conditions known to be related to ageing and cellular senescence, such as for example atherosclerosis, diabetes mellitus type 2, asthenia, and others.

The present invention relates to a cell culture medium for preparing liver, gastric, pancreatic, colon or intestinal adult stem cell isolated from adult tissue, as well as for maintaining such stem cell in the undifferentiated state. The cell culture medium comprises a base medium; an ABL and SRC dual kinase inhibitor/an ABL kinase inhibitor and a SRC kinase inhibitor; a mitogenic factor; a WNT signalling pathway activator; a stimulator for NAD+ and NADP+ generation; and a cAMP/P KA pathway activator. In a particular embodiment, the ABL and SRC dual kinase inhibitor is Dasatinib; the mitogenic factor is EGF; the WNT signalling pathway activator is R-Spondin 1; the stimulator for NAD+ and NADP+ generation is nicotinamide; and the cAMP/PKA pathway activator is cholera endotoxin.

Provided are a culture medium for expanding and cultivating human liver progenitor cells and an application thereof. The chemical components of the formula of the described culture medium are clear, no serum is present, and various components thereof cooperate with each other to synergize. The culture medium is used for the long-term expansion and cultivation of liver progenitor cells in vitro and is used for maintaining the dryness thereof, is beneficial in quickly and efficiently obtaining a large number of functional liver cells, and is suitable for clinical hepatocyte transplantation application as well as for the use of hepatocyte reactors in bioartificial livers.

The invention relates to differentiation methods for progenitor cells, e.g. mammalian epithelial stem cells, differentiation media for use in said methods, organoids and cells obtainable by said methods and uses, including therapeutic uses, thereof.

A method for producing a cell population containing liver progenitor cells, including the steps of

(1) preparing a culture substratum containing a cell population comprising liver progenitor cells and fibroblasts,
(2) physically removing the fibroblast colony from the culture substratum,
(3) detaching cells from the culture substratum, and recovering the detached cells, and
(4) culturing the cells recovered in the step (3) on a collagen-coated culture substratum, and recovering the cells not adhered to the culture substratum is provided by the present invention.