Media for Culturing, Preserving, and Administering Regenerative Cells

Abstract

A culture media, media supplement, or soluble matrix for cryopreservation or enhanced regenerative cell growth in culture and maintenance of multi-lineage differentiation potentiation. The inventive culture media, media supplement, or soluble matrix comprises a GAG composition comprising a sulfated GAG, such as chondroitin sulfate. A soluble matrix, a cell administration package or kit comprising the soluble matrix and a device for cell administration, and a method of use thereof, for administration of regenerative cells for treating a joint disease or other weakened or damaged tissue comprising the specified GAG compositions are further provided.

1 . A method of cryopreserving a regenerative cell comprising combining the regenerative cell with a glycosaminoglycan (GAG) composition at a temperature above 0° C., wherein at least one GAG is a sulfated GAG, and lowering the temperature to −18° C. or lower. 
     
     
         2 . The method of  claim 1 , wherein said GAG composition comprises hyaluronic acid, CS4 and CS6 chondroitin sulfate, and N-acetyl D-glucosamine at a concentration of 5-60% (v/v). 
     
     
         3 . The method of  claim 2 , wherein said composition is at a concentration of about 50% (v/v). 
     
     
         4 . The method of  claim 3 , wherein said composition is maintained at a temperature of −18° C. or lower for at least seven days. 
     
     
         5 . The method of  claim 1 , wherein said cell is an adipose-derived stem cell (ADSC). 
     
     
         6 . A method of enhancing an in vitro expansion rate of cells while maintaining multi-lineage differentiation potential, comprising culturing the cells in a cell culture media, treating the cells with a media supplement, or coating a surface on which the cells are growing with a matrix, wherein said cell culture media, media supplement, or matrix comprises a GAG composition, wherein at least one GAG is a sulfated GAG. 
     
     
         7 . The method of  claim 6 , wherein said GAG composition comprises hyaluronic acid, CS4 and CS6 chondroitin sulfate, and N-acetyl D-glucosamine at a concentration of 1-10% (v/v). 
     
     
         8 . The method of  claim 7 , wherein said GAG composition is at a concentration of 1-10% (v/v). 
     
     
         9 . The method of  claim 8 , wherein said composition is at a concentration of 3-7% (v/v). 
     
     
         10 . The method of  claim 9 , wherein said composition is at a concentration of 5% (v/v). 
     
     
         11 . The method of  claim 6 , wherein said cells are cultured with the culture media comprising the GAG composition. 
     
     
         12 . The method of  claim 6 , wherein said cells are treated with the media supplement comprising one or more GAG composition and then culturing the cells. 
     
     
         13 . The method of  claim 6 , wherein said cells are grown on a surface coated with a soluble matrix comprising the GAG composition. 
     
     
         14 . The method of  claim 6 , wherein said cell is an adipose-derived mesenchymal stem cell (Ad-MSC). 
     
     
         15 . A cryopreservation media or media supplement comprising a GAG composition for cryopreservation of cells and maintenance of multi-lineage differentiation potential comprising a sulfated GAG. 
     
     
         16 . The cryopreservation media or media supplement of  claim 15 , wherein said GAG composition comprises hyaluronic acid, CS4 and CS6 chondroitin sulfante and N-acetyl D-glucosamine at a concentration of 5-50% (v/v). 
     
     
         17 . The cryopreservation media or media supplement of  claim 15 , wherein said media or media supplement further comprises a carrier or soluble matrix for administration of the cells. 
     
     
         18 . The cryopreservation media or media supplement of  claim 15 , further comprising isolated regenerative cells. 
     
     
         19 . The cryopreservation media or media supplement of  claim 18 , wherein the cells are ADSCS. 
     
     
         20 . The cryopreservation media or media supplement of  claim 18  at a temperature of −80° C.