Composition for cryopreservation of cells and tissues

Abstract

A composition for cryopreservation of cells and tissues of human and other animals in a safe manner without using toxic substances such as DMSO, as well as for freeze preserving or freeze-drying of foods and pharmaceuticals. In examples, ε-poly-L-lysine is reacted with succinic anhydride so that 60% or more of amino groups are blocked; and, thus obtained polymer compound is added to Dulbecco-modified eagle MEM culture medium (DMEM) on market sale to form a cryopreservation liquid. In examples for foods or pharmaceuticals, the ε-poly-L-lysine derivative was added by 0.5-10 wt % to curb freeze concentration.

1 . A composition for cryopreservation of cells and tissues comprising: a polymer compound(s) comprised of a polymer(s) of units having amino groups, which is (are) selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, other poly-amino acids, amino-group-introduced polyshaccharides and polyallylamines; and
 physiological solution; and said polymer compound(s) being dissolved in the physiological solution by 1-50 w/w %. 
 
     
     
         2 . The composition according to  claim 1 , wherein 50-99 mol % of amino groups of said polymer compound(s) is blocked by being reacted with carboxylic anhydride to have carboxylic groups or acetyl groups. 
     
     
         3 . A composition for cryopreservation of cells and tissues comprising: a polymer compound(s) comprised of a polymer(s) of units having carboxyl, groups, which is (are) selected from a group consisting of polyacrylic acid, α-poly-glutamic acid, γ-poly-glutamic acid, poly-glutamic acid, poly-aspartic acid, other poly-amino acids and carboxylated polyshaccharides; and physiological solution; and said polymer compound(s) being dissolved in the physiological solution by 1-50 w/w %. 
     
     
         4 . The composition according to  claim 3 , wherein 1-50 mol % of amino groups of said polymer compound(s) is blocked by being reacted with a compound(s) having a plurality of amino groups such as diamines, triamines and polyamines. 
     
     
         5 . The composition according to  claim 1 , wherein the physiological solution is a saline, Dulbecco-modified eagle MEM culture medium (DMEM), or a culture medium for cells or tissues. 
     
     
         6 . The composition according to  claim 1 , wherein said polymer compound is ε-poly-L-lysine having number-average molecular weight in a range of 1000-20,000. 
     
     
         7 . The composition according to  claim 1 , wherein said polymer compound is polyacrylic acid having number-average molecular weight in a range of 1000-3,000,000. 
     
     
         8 . An additive composition for freeze preservation or freeze-drying of foods or pharmaceuticals, comprising: a polymer compound(s) comprised of a polymer(s) of units having amino groups, which is (are) selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, other poly-amino acids, amino-group-introduced polyshaccharides and polyallylamines; wherein amino groups of said polymer compound(s) is blocked by being reacted with carboxylic anhydride to have carboxylic groups or acetyl groups. 
     
     
         9 . A method of manufacturing foods pharmaceuticals comprising adding a polymer compound(s) comprised of a polymer(s) of units having amino groups, which is (are) selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, other poly-amino acids, amino-group-introduced polyshaccharides and polyallylamines, to water-bearing material(s) for producing the foods or the pharmaceuticals so that concentration of said polymer compound in the water-bearing material(s) becomes in a range of 1-15 w/w %. 
     
     
         10 . The method of manufacturing foods or pharmaceuticals according to  claim 9 , wherein 50-99 mol % of amino groups of said polymer compound(s) is blocked by being reacted with carboxylic anhydride to have carboxylic groups or acetyl groups. 
     
     
         11 . The method of manufacturing foods or pharmaceuticals according to  claim 9 , wherein said polymer compound is ε-poly-L-lysine having number-average molecular weight in a range of 1000-20,000. 
     
     
         12 . A method of manufacturing fuel cells comprising:
 preparing a polymer compound(s) comprised of a polymer(s) of units having amino groups, which is (are) selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, other poly-amino acids, amino-group-introduced polyshaccharides and polyallylamines; in which 50-99 mol % of amino groups of said polymer compound(s) have been blocked by being reacted with succinic anhydride, acetic anhydride or other carboxylic anhydride to have carboxylic groups or acetyl groups; and   adding said polymer compound to a material for coating film covering surface of separator or solid electrolyte membrane, or for other surface layer exposed to insides of the fuel cells so that concentration of said polymer compound in said material becomes in a range of 1-15 w/w %.   
     
     
         13 . The method of manufacturing fuel cells according to  claim 12 , wherein said polymer compound is ε-poly-L-lysine having number-average molecular weight in a range of 1000-20,000. 
     
     
         14 . The method of manufacturing fuel cells according to  claim 12  or  13 , wherein the material for the coating film is UV curable resin liquid. 
     
     
         15 . A method of manufacturing fuel cells or other industrial products comprising:
 preparing a polymer compound(s) comprised of a polymer(s) of units having amino groups, which is (are) selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, other poly-amino acids, amino-group-introduced polyshaccharides and polyallylamines; in which 50-99 mol % of amino groups of said polymer compound(s) have been blocked by being reacted with succinic anhydride, acetic anhydride or other carboxylic anhydride to have carboxylic groups or acetyl groups; and   adding said polymer compound to a material for forming a hydrophilic surface layer on an inner element of the fuel cell or other industrial product so that concentration of said polymer compound in said material becomes in a range of 1-15 w/w %.   
     
     
         16 . The composition according to  claim 3 , wherein the physiological solution is a saline, Dulbecco-modified eagle MEM culture medium (DMEM), or a culture medium for cells or tissues. 
     
     
         17 . The composition according to  claim 3 , wherein said polymer compound is ε-poly-L-lysine having number-average molecular weight in a range of 1000-20,000. 
     
     
         18 . The composition according to  claim 3 , wherein said polymer compound is polyacrylic acid having number-average molecular weight in a range of 1000-3,000,000.