Polymerized Hemoglobin Media and its Use in Isolation and Transplantation of Islet Cells

Abstract

Solutions and suspensions comprising polymerized hemoglobin derived from human blood are disclosed. The solutions and suspensions may comprise cell culture medium, an enzyme (such as a protease), and/or a buffer. Processes of preparing the solutions and suspensions are also disclosed. The solutions and suspensions may be employed in methods of isolating mammalian cells, such as pancreatic islets, methods of preserving mammalian tissue and organs, methods of aiding the recovery of mammalian cells following their isolation, methods of maintaining mammalian cells, methods of propagating mammalian cells, and methods of treating a mammal with diabetes.

1 . A solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) an enzyme.  
     
     
         2 . The solution of  claim 1 , wherein the enzyme is a protease.  
     
     
         3 . The solution of  claim 2 , wherein the protease is collagenase.  
     
     
         4 . The solution of  claim 1 , further comprising cell culture medium.  
     
     
         5 . The solution of  claim 4 , wherein the cell culture medium is RPMI.  
     
     
         6 . The solution of  claim 5 , wherein the enzyme is collagenase.  
     
     
         7 . The solution of  claim 1 , further comprising a buffer.  
     
     
         8 . The solution of  claim 1 , wherein the polymerized hemoglobin is derived from human blood and is pyridoxylated.  
     
     
         9 . The solution of  claim 8 , further comprising cell culture medium.  
     
     
         10 . The solution of  claim 9 , wherein the enzyme is collagenase and the cell culture medium is RPMI.  
     
     
         11 . The solution of  claim 1 , wherein the solution is oxygenated.  
     
     
         12 . The solution of  claim 4 , wherein the solution is oxygenated.  
     
     
         13 . The solution of  claim 10 , wherein the solution is oxygenated.  
     
     
         14 . A solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.  
     
     
         15 . The solution of  claim 14 , wherein the cell culture medium is RPMI.  
     
     
         16 . The solution of  claim 14 , wherein the solution is oxygenated.  
     
     
         17 . A process for the preparation of the solution of  claim 1 , comprising mixing (a) polymerized hemoglobin derived from mammalian and (b) one or more enzymes.  
     
     
         18 . A process for the preparation of the solution of  claim 2 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) a protease.  
     
     
         19 . A process for the preparation of the solution of  claim 3 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) collagenase.  
     
     
         21 . A process for the preparation of the solution of  claim 4 , comprising mixing (a) polymerized hemoglobin derived from human blood, (b) one or more enzymes, and (c) cell culture medium.  
     
     
         21 . A process for the preparation of the solution of  claim 5 , comprising mixing (a) polymerized hemoglobin derived from human blood, (b) one or more enzymes, and (c) RPMI.  
     
     
         22 . A process for the preparation of the solution of  claim 6 , comprising mixing (a) polymerized hemoglobin derived from human blood, (b) collagenase, and (c) RPMI.  
     
     
         23 . A process for the preparation of the solution of  claim 7 , comprising mixing (a) polymerized hemoglobin derived from human blood, (b) one or more enzymes, and (c) a buffer.  
     
     
         24 . A process for the preparation of the solution of  claim 8 , comprising mixing (a) polymerized and pyridoxylated hemoglobin derived from human blood and (b) one or more enzymes.  
     
     
         25 . A process for the preparation of the solution of  claim 9 , comprising mixing polymerized and pyridoxylated hemoglobin derived from human blood, (b) one or more enzymes, and (c) cell culture medium.  
     
     
         26 . A process for the preparation of the solution of  claim 10 , comprising mixing polymerized and pyridoxylated hemoglobin derived from human blood, (b) collagenase, and (c) RPMI.  
     
     
         27 . A process for the preparation of the solution of  claim 11 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) one or more enzymes, followed by contacting the mixture with O 2  gas.  
     
     
         28 . A process for the preparation of the solution of  claim 12 , comprising mixing (a) polymerized hemoglobin derived from human blood, (b) one or more enzymes, and (c) cell culture medium, followed by contacting the mixture with O 2  gas.  
     
     
         29 . A process for the preparation of the solution of  claim 13 , comprising mixing (a) polymerized and pyridoxylated hemoglobin derived from human blood, (b) collagenase, and (c) RPMI, followed by contacting the mixture with O 2  gas.  
     
     
         30 . A process for the preparation of the solution of  claim 14 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) cell culture medium.  
     
     
         31 . A process for the preparation of the solution of  claim 15 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) RPMI.  
     
     
         32 . A process for the preparation of the solution of  claim 16 , comprising mixing (a) polymerized hemoglobin derived from human blood and (b) cell culture medium, followed by contacting the mixture with O 2  gas.  
     
     
         33 . A methods of preserving a whole mammalian organ, comprising contacting the whole organ with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.  
     
     
         34 . A method according to  claim 33 , where the organ is a human liver, a human lung, or a human kidney.