Method for producing adult liver progenitor 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.
1. An isolated, human adult liver progenitor cell wherein said cell is measured:
(a) positive for α-smooth muscle actin (ASMA), albumin (ALB), and CD140b; and
(b) negative for Sushi domain containing protein 2 (SUSD2) and Cytokeratin-19 (CK-19).
2. The cell of claim 1 wherein said cell is further measured positive for:
(a) At least one hepatic marker selected from the group consisting of albumin, HNF-3B, HNF-4, CYP1A2, CYP2C9, CYP2E1 and CYP3A4;
(b) At least one mesenchymal marker selected from the group consisting of Vimentin, CD90, CD73, CD44, and CD29;
(c) At least one liver-specific activity selected from the group consisting of urea secretion, bilirubin conjugation, alpha-1-antitrypsin secretion, and CYP3A4 activity;
(d) At least one marker selected from the group consisting of ATP2B4, ITGA3, TFRC, SLC3A2, CD59, ITGB5, CD151, ICAM1, ANPEP, CD46, and CD81; and
(e) At least one marker selected from the group consisting of MMP1, ITGA11, FMOD, KCND2, CCL11, ASPN, KCNK2, and HMCN1.
3. The cell of claim 1 characterized in that the cell is measured negative for:
(a) CD271;
(b) At least one marker selected from the group consisting of ITGAM, ITGAX, IL1R2, CDH5, and NCAM1; and
(c) At least one marker selected from the group consisting of HP, CP, RBP4, APOB, LBP, ORM1, CD24, CPM, and APOC1.
4. The cell of claim 1 wherein said cell is further measured positive for at least one liver-specific activity selected from the group consisting of sulfotransferase activity, tryptophan-2,3-dioxygenase activity, liver carboxylase activity, ammonia metabolism, and glycogen storage.
5. An isolated cell population comprising at least 60%, or between 60% and 99%, or between 70% and 90%, of the adult liver progenitor cells according to claim 1 .
6. The isolated cell population of claim 5 wherein said cell population is further differentiated into cells presenting liver-specific activities.
7. The adult liver progenitor cell of claim 1 , or an isolated cell population comprising at least 60% of said cells, wherein said adult liver progenitor cell or isolated cell population is further modified by means of one or more chemical agents, cell culture medium, growth factors, and/or nucleic acid vectors.
8. A pharmaceutical composition comprising the adult liver progenitor cell of claim 1 , or an isolated cell population comprising at least 60% of said cells.
9. A method of treating a liver disease, the method comprising administering to a subject in need thereof, a pharmaceutical composition comprising a therapeutically effective amount of the adult liver progenitor cells according to claim 1 or an isolated cell population comprising at least 60% of said adult liver progenitor cells.
10. The method of claim 9 wherein the liver disease is an inborn error of liver metabolism, an inherited Blood Coagulation Disorder, progressive familial intrahepatic cholestasis type 1 / 2 / 3, alpha 1-Antitrypsin Deficiency, defect of liver cell transporters, Porphyria, fatty liver or fibrotic liver disease, primary biliary cirrhosis, sclerosing cholangitis, liver degenerative disease, or acute or chronic liver failure.
11. A kit comprising the isolated, human adult liver progenitor cell according to claim 1 , or an isolated cell population comprising at least 60% of said cells.
12. The kit of claim 11 wherein said kit further comprises one or more vials containing said isolated cell population and one or more of the following elements: a device, a disposable material, a solution, a chemical product, a biological product, and/or instructions for using elements of said kit.