Protection of Progenitor Cells and Regulation of Their Differentiation

Abstract

The present invention relates to the use of polysulfated polysaccharides in combination with progenitor cells to improve the viability of the progenitor cells including improving the cryopreservation of the progenitor cells and provides novel compositions, methods and uses. The present invention also relates to the use of polysulfated polysaccharides to regulate the proliferation and differentiation of progenitor cells.

1 . A composition comprising progenitor cells together with a polysulfated polysaccharide or biologically active molecular fragment thereof. 
     
     
         2 . The composition of  claim 1  further comprising a carrier medium. 
     
     
         3 . The composition of  claim 2 , wherein the carrier medium is a culture medium, cryopreservation medium and/or a pharmaceutically acceptable carrier. 
     
     
         4 . The composition of  claim 1  wherein the progenitor cell is a chondroprogenitor cell. 
     
     
         5 . The composition of  claim 1  wherein the progenitor cell is a Stro-1 bri  cell, and/or a Stro-1 bri  progeny cell. 
     
     
         6 . The composition of  claim 1  wherein the polysulfated polysaccharide is selected from pentosan polysulfate chondroitin polysulfate, chitosan polysulfate, dextran polysulfate and heparin (high and low molecular weight) together with their pharmaceutically acceptable salts. 
     
     
         7 . The composition of  claim 1  wherein the polysulfated polysaccharide is selected from pentosan polysulfate, the sodium salt of pentosan polysulfate (NaPPS), the magnesium salt of pentosan polysulfate (MgPPS), and/or the calcium salt of pentosan polysulfate (CaPPS). 
     
     
         8 . The composition of  claim 1  wherein the composition comprises about 1000 to about 1×10 10  cells; about 1×10 5 -1×10 9  cells; about 100,000 to about 5×10 8  cells; about 500,000 to about 2×10 8  cells, about 1×10 6  to about 2×10 8  cells; about 1×10 6  to about 1×10 8  cells; about 1×10 6 , 2×10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×10 6 , 7×10 6 , 8×10 6 , 9×10 6 , 1×10 7 , 2×10 7 , 3×10 7 , 4×10 7 , 5×10 7 , 6×10 7 , 7×10 7 , 8×10 7 , 9×10 7 , 1×10 8 , 2×10 8 , 3×10 8 , 4×10 8 , 5×10 8 , 6×10 8 , 7×10 8 , 8×10 8 , or about 9×10 8  cells. 
     
     
         9 . The composition of  claim 1  wherein the composition comprises polysulfated polysaccharide in a concentration of about 500 ng/ml/million cells-10 mg/ml/million cells, 500 ng/ml/million cells-2000 μg/ml/million cells, 1 μg/ml/million cells-1000 μg/ml/million cells, or 1 μg/ml/million cells-500 μg/ml/million cells, 500 ng-10 μg/ml/million cells, 1 μg-10 μg/ml/million cells, 1 μg-8 μg/ml/million cells, 1 μg-6 μg/ml/million cells, 1 μg-5 μg/ml/million cells, 1 μg-3 μg/ml/million cells, 2 μg-6 μg/ml/million cells, 2.5 μg-5 μg/ml/million cells, or 3 μg-5 μg/ml/million cells, 1 μg-100 μg/ml/million cells, 1 μg-50 μg/ml/million cells, 1 μg-20 μg/ml/million cells, 1 μg-15 μg/ml/million cells, 10 μg-100 μg/ml/million cells, 20 μg-100 μg/ml/million cells, or 50 μg-100 μg/ml/million cells, 1 μg-1000 μg/ml/million cells, 100 μg-800 μg/ml/million cells, 100 μg-600 μg/ml/million cells, 100 μg-500 μg/ml/million cells, 200 μg-500 μg/ml/million cells, 500 ng, 1 μg, 2 μg, 2.5 μg, 5 μg, 10 μg, 15 μg, 20 μg, 30 μg, 40 μg, 50 μg, 60 μg, 70 μg, 80 μg, 90 μg, 100 μg, 150 μg, 200 μg, 250 μg, 300 μg, 350 μg, 400 μg, 450 μg, 500 μg, 550 μg, 600 μg, 650 μg, 700 μg, 750 μg, 800 μg, 850 μg, 900 μg, 950 μg, 1000 μg, 1050 μg, 1100 μg, 1150 μg, 1200 μg, 1250 μg, 1300 μg, 1350 μg, 1400 μg, 1450 μg, 1500 μg, 1550 μg, 1600 μg, 1650 μg, 1700 μg, 1750 μg, 1800 μg, 1850 μg, 1900 μg, 1950 μg, or 2000 μg/ml/million cells. 
     
     
         10 . The composition of  claim 1  wherein the composition comprises about 1×10 6 -1×10 8  progenitor cells and 25-50 mg polysulfated polysaccharide; 1×10 8  progenitor cells and 25-50 mg/ml polysulfated polysaccharide. 
     
     
         11 . The composition of  claim 1  wherein the composition further comprises one or more of the following components:
 organic and/or inorganic salts; 
 buffers 
 proteins such as BSA or transferin; 
 growth factors and cytokines, including insulin like growth factor, insulin, fibroblast like growth factors; BMP-TGF-beta super family (eg, BMP-2, BMP-7, BMP-8, TGF beta) and fibroblast growth factor family, IGF, FGF, EGF, PDGF, VEGF; 
 animal sera including FBS, new-born calf, all other mammalian species; 
 cryopreservation agents such as Profreeze® and CryoStor®; 
 cryoprotectorants, including dimethyl sulfoxide (DMSO), glycerol, trehalose, sucrose and other sugars or dimethylacetamide; 
 carbohydrates; 
 vitamins/co-factors; 
 hormones 
 antibiotics 
 attachment factors; 
 amino acids; 
 plasma expanders like dextran; 
 plasma both human and other mammalian species; 
 plasma substitute; 
 hyaluronan and/or hyaluronic acid, both natural or cross linked. 
 
     
     
         12 . The composition of  claim 11  wherein the DMSO is present in the range of 1-20%, 1-15%, 5-15%, 1-10%, 5%, 7.5%, 10%, 15% or 20%; and/or the FBS is in the range of 1-50%, 1-20%, 1-10%, 5%, 7.5%, 10%, 15% or 20%. 
     
     
         13 . The composition of  claim 1  wherein the carrier medium comprises Profreeze®. 
     
     
         14 . The composition of  claim 1  wherein the composition further comprises NC4 or a biologically active fragment thereof. 
     
     
         15 . A method of enhancing cryopreservation of progenitor cells, comprising exposing the progenitor cells to a polysulfated polysaccharide or a biologically active molecular fragment thereof. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . A method of regulating the proliferation of progenitor cells, comprising exposing a polysulfated polysaccharide or biologically active molecular fragment thereof to a progenitor cell. 
     
     
         19 . (canceled) 
     
     
         20 . A method of regulating differentiation of progenitor cells by exposing a polysulfated polysaccharide or biologically active molecular fragment thereof to a progenitor cell, including differentiation into chondrocytes, differentiation into osteoblasts or differentiation into adipocytes. 
     
     
         21 - 22 . (canceled)