Directly injectable formulations which provide enhanced cryoprotection of cell products

Abstract

This invention provides compositions and methods for cryoprotection of recombinant live cancer cells. Specifically, an improved cryoprotective medium is provided which includes a hydroxyethyl starch and/or derivative thereof alone or in combination with either DMSO or glycerol.

1 . A composition comprising a recombinant live cancer cell in a cryoprotective medium, wherein the cryoprotective medium comprises about 5% by weight to about 22% by weight of a hydroxyethyl starch (HESS) or derivative thereof and about 1% by weight to about 15% by weight of DEMOS.  
     
     
         2 . The composition of  claim 1 , further comprising from about 0% by weight to about 10% by weight human serum albumin (HAS).  
     
     
         3 . The composition of  claim 2 , wherein the HESS comprises HESS: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is from about 0.60-0.80;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from about 0.40-0.60; or a combination of (a) and (b).    
     
     
         4 . The composition of  claim 3 , wherein the HESS comprises HESS: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is about 0.70;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from 0.4 to about 0.5; or a combination of (a) and (b).    
     
     
         5 . The composition of  claim 2 , wherein the HES comprises HES with: 
 (a) a molecular weight of from about 420 to about 480 kDa;    (b) a molecular weight of from about 200 to about 290 kDa; or a combination of (a) and (b).    
     
     
         6 . The composition of  claim 2 , wherein the recombinant live cancer cell comprises an introduced heterologous nucleic acid coding sequence for a cytokine or costimulatory molecule.  
     
     
         7 . The composition of  claim 6 , wherein the cytokine is granulocyte-macrophage colony stimulating factor (GM-CSF).  
     
     
         8 . The composition of  claim 2 , wherein the cancer cell is selected from the group consisting of a lung cancer cell, a pancreatic cancer cell, a prostate cancer, a kidney cancer cell, a myeloma cell, and a leukemic cell.  
     
     
         9 . The composition of  claim 2 , wherein the recombinant live cancer cell composition comprises patient-derived tumor cells.  
     
     
         10 . The composition of  claim 9 , wherein said recombinant live cancer cell is autologous.  
     
     
         11 . The composition of  claim 2 , wherein said recombinant live cancer cell is allogeneic.  
     
     
         12 . The composition of  claim 2 , wherein said recombinant live cancer cell is a bystander cell.  
     
     
         13 . A composition comprising a recombinant live cancer cell in a cryoprotective medium, wherein the cryoprotective medium comprises about 8% by weight HES, about 2% by weight HSA, and about 5% by weight DMSO.  
     
     
         14 . A composition comprising a recombinant live cancer cell in a cryoprotective medium, wherein the cryoprotective medium comprises about 5% by weight to about 22% by weight of the a hydroxyethyl starch (HES) or derivative thereof and about 1% by weight to about 15% by weight of the glycerol, said composition further comprising from about 0% to about 10% human serum albumin (HSA).  
     
     
         15 . The composition of  claim 14 , wherein the HES comprises HES: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is from about 0.60-0.80;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from about 0.40-0.60; or a combination of (a) and (b).    
     
     
         16 . The composition of  claim 15 , wherein the HES comprises HES: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is about 0.70;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from 0.4 to about 0.5; or a combination of (a) and (b).    
     
     
         17 . The composition of  claim 14 , wherein the HES comprises HES with: 
 (a) a molecular weight of from about 420 to about 480 kDa;    (b) a molecular weight of from about 200 to about 290 kDa; or a combination of (a) and (b).    
     
     
         18 . The composition of  claim 14 , wherein the recombinant live cancer cell comprises an introduced heterologous nucleic acid coding sequence for a cytokine or costimulatory molecule.  
     
     
         19 . The composition of  claim 18 , wherein the cytokine is granulocyte-macrophage colony stimulating factor (GM-CSF).  
     
     
         20 . The composition of  claim 14 , wherein the cancer cell is selected from the group consisting of a lung cancer cell, a pancreatic cancer cell, a prostate cancer cell, a kidney cancer cell, a myeloma cell, and a leukemic cell.  
     
     
         21 . The composition of  claim 14 , wherein said recombinant live cancer cell composition comprises patient-derived tumor cells.  
     
     
         22 . The composition of  claim 21 , wherein said recombinant live cancer cell is autologous.  
     
     
         23 . The composition of  claim 14 , wherein said recombinant live cancer cell is allogeneic.  
     
     
         24 . The composition of  claim 14 , wherein said recombinant live cancer cell is a bystander cell.  
     
     
         25 . A composition comprising a recombinant live cancer cell in a cryoprotective medium, wherein the cryoprotective medium comprises about  8 % by weight HES, about 2% by weight HSA, and about 5% by weight glycerol.  
     
     
         26 - 44 . (canceled)  
     
     
         45 . A kit for preserving cell viability comprising: a container containing a cryoprotective medium comprising about 5% by weight to about 22% by weight hydroxyethyl starch (HES) or derivative thereof alone or in combination with about 1% by weight to about 15% by weight DMSO; from about 0% to about 10% human serum albumin (HSA), and instructional materials teaching the use of the cryoprotective medium to preserve cell viability.  
     
     
         46 . The kit of  claim 45 , wherein the HES comprises HES: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is about 0.70;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from 0.4 to about 0.5; or a combination of (a) and (b).    
     
     
         47 . The kit of  claim 45 , wherein the HES comprises HES with: 
 (a) a molecular weight of from about 420 to about 480 kDa;    (b) a molecular weight of from about 200 to about 290 kDa; or a combination of (a) and (b).    
     
     
         48 . A kit for preserving cell viability comprising: a container containing a cryoprotective medium comprising about 5% by weight to about 22% by weight hydroxyethyl starch (HES) or derivative thereof, about 3% by weight to about 15% by weight glycerol; from about 0% to about 10% human serum albumin (HSA), and, instructional materials teaching the use of the cryoprotective medium to preserve cell viability.  
     
     
         49 . The kit of  claim 48 , wherein the HES comprises HES: 
 (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is about 0.70;    (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from 0.4 to about 0.5; or a combination of (a) and (b).    
     
     
         50 . The kit of  claim 48 , wherein the HES comprises HES with: 
 (a) a molecular weight of from about 420 to about 480 kDa;    (b) a molecular weight of from about 200 to about 290 kDa; or a combination of (a) and (b).