Conditioned medium from human adult liver stem cells and its use in the treatment of liver disorders

Abstract

The invention relates to cell-free compositions obtained by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in cell culture medium and isolating the resulting conditioned medium (ADHLSC-CM) that has advantageous property ties, such as anti-fibrotic effects. ADHLSC-CM, compositions based on ADHLSC-CM, and other related and derived products, can be used in cell culture processes or as a medicament, more particularly for the treatment of diseases involving organ injury, organ failure, in organ or cell transplantation or the pathological disruption, inflammation, degeneration, and/or proliferation of cells within a tissue or an organ, in particular within liver.

1 . A cell-free conditioned medium that contains soluble proteins and/or micro vesicles and that is obtainable by culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells. 
     
     
         2 . The cell-free conditioned medium according to  claim 1 , wherein the medium is serum-free. 
     
     
         3 . A cell-free composition obtainable by fractioning the cell-free conditioned medium according to  claim 1 , wherein said fractioning comprises filtering, enzymatically digesting, centrifuging, adsorbing, and/or separating by chromatography the cell-free conditioned medium. 
     
     
         4 . The cell-free conditioned medium according to  claim 1 , wherein adult-derived human liver stem/progenitor cells (ADHLSC) co-express at least one mesenchymal marker selected from CD90, CD73, CD44, vimentin and a-smooth muscle actin with at least an hepatic marker selected from albumin, CD29, alpha-fetoprotein, alpha-1 antitrypsin, HNF-4 and MRP2 transporter. 
     
     
         5 . The cell-free conditioned medium according to  claim 1 , wherein adult-derived human liver stem/progenitor cells (ADHLSC) are albumin-positive, vimentin-positive, alpha smooth muscle actin-positive, cytokeratin-19-negative, and CD133-negative. 
     
     
         6 . The cell-free conditioned medium according to  claim 1 , comprising at least one of soluble proteins selected from the group consisting of Hepatocyte Growth Factor (HGF), Vascular Endothelial Growth Factor (VEGF), Eotaxin (CCL11), Interleukin-6 (IL-6), and Interleukin-8 (IL-8). 
     
     
         7 . The cell-free conditioned medium or the cell-free composition according to  claim 6 , wherein all of said soluble proteins are present at a concentration of at least 1 ng/ml. 
     
     
         8 . The cell-free conditioned medium according to  claim 1 , wherein said microvesicles are smaller than 0.40 μm. 
     
     
         9 . The cell-free conditioned medium according to  claim 1  that is concentrated at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 50-fold, or at least about 100-fold. 
     
     
         10 . The cell-free conditioned medium according to  claim 1  that is diluted at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 50-fold, or at least about 100-fold. 
     
     
         11 . A method for producing a cell-free conditioned medium comprising the steps of culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells. 
     
     
         12 . The method for producing a cell-free conditioned medium according to  claim 11 , wherein:
 (a) the cell culture medium is a serum-free medium; and/or   (b) the cell culture medium is separated from ADHLSC after culturing ADHLSC in the cell culture medium for at least 2 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, or at least 24 hours; and/or   (c) adult-derived human liver stem/progenitor cells (ADHLSC) co-express at least one mesenchymal marker selected from CD90, CD73, CD44, vimentin and a-smooth muscle actin with at least an hepatic marker selected from albumin, CD29, alpha-fetoprotein, alpha-1 antitrypsin, HNF-4 and MRP2 transporter.   
     
     
         13 . A method for producing a cell-free composition comprising Hepatocyte Growth Factor (HGF), Vascular Endothelial Growth Factor (VEGF), Eotaxin (CCL11), Interleukin-6 (IL-6), and Interleukin-8 (IL-8) at a concentration of at least 1 ng/ml comprising the steps of culturing adult-derived human liver stem/progenitor cells (ADHLSC) in a cell culture medium and separating the cell culture medium from the cells. 
     
     
         14 . A method of treating a disorder in a subject in need of said treatment comprising the administration of a therapeutically or prophylactically effective amount of the cell-free conditioned medium of  claim 1  to the subject. 
     
     
         15 . A pharmaceutical formulation comprising a pharmaceutically effective amount of the cell-free conditioned medium of  claim 1 . 
     
     
         16 . The pharmaceutical formulation according to  claim 15 , further comprising one or more exogenous active ingredients. 
     
     
         17 . The method of  claim 14 , wherein the disorder is a fibrotic disorder. 
     
     
         18 . The method of  claim 14 , wherein the disorder is a liver disorder. 
     
     
         19 . The method of  claim 14 , wherein the subject is suffering from organ injury, organ failure, or is in need of organ or cell transplantation. 
     
     
         20 . A method of treating a disorder in a subject in need of said treatment comprising the administration of a therapeutically or prophylactically effective amount of the cell-free composition of  claim 3  to the subject.