Solutions for increasing the stability and shelf life of an organ and tissue preservation solution

Abstract

Organ and tissue preservation solutions having improved stability are disclosed. The solutions are comprised of two separate solutions. The first solution, Solution A, is comprised of a balanced salt solution that is stable in solution at a pH of 7.0 or above. A second solution, Solution B, is comprised of an aqueous solution containing L-glutathione and/or cysteinylglycine, a sugar such as D-glucose, L-Arginine, a reducing agent such as ascorbic acid and water at a pH of below 7.0, preferably from about 3.0 to 5.0. The two Solutions are then mixed together at the point of use and the pH adjusted to about 7.3 resulting in the organ and tissue preservation solution having improved stability. Preferably, solution A has a pH of about 7.6 and solution B has a pH of about 5.0. The present invention is also comprised of kits 20 that contain the two Solutions in two separate containers 22, 24. In an alternate embodiment, the sugar can be in Solution A.

1 - 30 . (canceled) 
     
     
         31 . A method for preparing an organ or tissue preservation solution, comprising:
 preparing a first precursor aqueous solution comprising a balanced salt solution, and having a pH of about 7.4 to 8.0, wherein the first precursor aqueous solution does not include L-Arginine, reduced glutathione, or ascorbic acid;   preparing a second precursor solution comprising water, sugar, L-Arginine, reduced glutathione, ascorbic acid and, optionally, cysteinylglycine, and having a pH of about 3.0 to 4.0; and   mixing the first precursor aqueous solution together with the second precursor solution at a point of use to form the organ and tissue preservation solution,   wherein the organ and tissue preservation solution has a pH of about 7.3.   
     
     
         32 . A method for preserving a tissue or organ, comprising:
 preparing the organ and tissue preservation solution according to  claim 31 ; and   bringing a tissue or organ into contact with the organ and tissue preservation solution.   
     
     
         33 . The method of  claim 32 , wherein the tissue or organ is selected from the group consisting of saphenous veins, epigastric arteries, gastroepiploic arteries, radial arteries, heart, lungs, kidney, brain, muscle grafts, skin, intestine, bone, appendages, eyes, and portions of the tissue or organs. 
     
     
         34 . The method of  claim 31 , wherein the sugar is D-Glucose. 
     
     
         35 . The method of  claim 31 , wherein the second precursor solution comprises cysteinylglycine. 
     
     
         36 . The method of  claim 31 , wherein the balanced salt solution comprises:
 calcium chloride dihydrate in a concentration of about 0.14 g/L;   potassium chloride in a concentration of about 0.4 g/L;   potassium phosphate monobasic in a concentration of about 0.06 g/L;   magnesium chloride hexahydrate in a concentration of about 0.1 g/L;   magnesium sulfate heptahydrate in a concentration of about 0.1 g/L sodium chloride in a concentration of about 8 g/L;   sodium bicarbonate in a concentration of about 0.36 g/L; and   sodium phosphate dibasic heptahydrate in a concentration of about 0.03 g/L.   
     
     
         37 . The method of  claim 31 , wherein the organ and tissue preservation solution comprises the ascorbic acid in a concentration of about 25-1000 μM, the glutathione in a concentration of about 50-2000 μM, the L-arginine in a concentration of about 250-2000 μM, and the sugar in a concentration of about 5.55 mM. 
     
     
         38 . A method for preparing an organ or tissue preservation solution, comprising:
 preparing a first precursor aqueous solution comprising a balanced salt solution, and having a pH of about 7.4 to 8.0, wherein the first precursor aqueous solution does not include L-Arginine, reduced glutathione, or ascorbic acid;   preparing a second precursor solution comprising water, L-Arginine, ascorbic acid, and reduced glutathione, and having a pH of about 3.0 to 4.0; and   mixing the first precursor aqueous solution together with the second precursor solution at a point of use to form the organ and tissue preservation solution,   wherein the organ and tissue preservation solution has a pH of about 7.3, and   wherein one of the first and second precursor aqueous solutions includes a sugar.   
     
     
         39 . A method for preserving a tissue or organ, comprising:
 preparing the organ and tissue preservation solution according to  claim 38 ; and   bringing a tissue or organ into contact with the organ and tissue preservation solution.   
     
     
         40 . The method of  claim 39 , wherein the tissue or organ is selected from the group consisting of saphenous veins, epigastric arteries, gastroepiploic arteries, radial arteries, heart, lungs, kidney, brain, muscle grafts, skin, intestine, bone, appendages, eyes, and portions of the tissue or organs. 
     
     
         41 . The method of  claim 38 , wherein the balanced salt solution comprises:
 calcium chloride dihydrate in a concentration of about 0.14 g/L;   potassium chloride in a concentration of about 0.4 g/L;   potassium phosphate monobasic in a concentration of about 0.06 g/L;   magnesium chloride hexahydrate in a concentration of about 0.1 g/L;   magnesium sulfate heptahydrate in a concentration of about 0.1 g/L sodium chloride in a concentration of about 8 g/L;   sodium bicarbonate in a concentration of about 0.36 g/L; and   sodium phosphate dibasic heptahydrate in a concentration of about 0.03 g/L.   
     
     
         42 . The method of  claim 38 , wherein the organ and tissue preservation solution comprises the ascorbic acid in a concentration of about 25-1000 μM, the glutathione in a concentration of about 50-2000 μM, the L-arginine in a concentration of about 250-2000 μM, and the sugar in a concentration of about 5.55 mM. 
     
     
         43 . The method claimed in  claim 38 , wherein the sugar is glucose. 
     
     
         44 . The method of  claim 38 , wherein the first precursor solution comprises the sugar. 
     
     
         45 . The method of  claim 38 , wherein the second precursor solution comprises the sugar. 
     
     
         46 . A method of preserving living tissue, comprising:
 combining a first precursor aqueous solution with a second precursor aqueous solution to form an organ and tissue preservation solution; and   contacting the living tissue with the organ and tissue preservation solution;   wherein the first precursor aqueous solution comprises a balanced salt solution, has a pH of about 7.4 to 8.0, and does not include reduced glutathione, L-Arginine or ascorbic acid,   wherein the second precursor aqueous solution comprises water, reduced glutathione, ascorbic acid, L-Arginine, and optionally, cysteinylglycine, at a pH of about 3.0 to 4.0,   wherein the organ and tissue preservation solution has a pH of about 7.3, and   wherein one of the first and second precursor aqueous solutions comprises a sugar.   
     
     
         47 . The method of  claim 46 , wherein the sugar is glucose. 
     
     
         48 . The method of  claim 46 , wherein the first precursor solution comprises the sugar. 
     
     
         49 . The method of  claim 46 , wherein the second precursor solution comprises the sugar. 
     
     
         50 . The method of  claim 46 , wherein the second precursor aqueous solution comprises cysteinylglycine. 
     
     
         51 . The method of  claim 46 , wherein the organ and tissue preservation solution comprises the ascorbic acid in a concentration of about 25-1000 μM, the glutathione in a concentration of about 50-2000 μM, the L-arginine in a concentration of about 250-2000 μM, and the sugar in a concentration of about 5.55 mM. 
     
     
         52 . The method of  claim 46 , wherein the balanced salt solution comprises:
 calcium chloride dihydrate in a concentration of about 0.14 g/L;   potassium chloride in a concentration of about 0.4 g/L;   potassium phosphate monobasic in a concentration of about 0.06 g/L;   magnesium chloride hexahydrate in a concentration of about 0.1 g/L;   magnesium sulfate heptahydrate in a concentration of about 0.1 g/L   sodium chloride in a concentration of about 8 g/L;   sodium bicarbonate in a concentration of about 0.36 g/L; and   sodium phosphate dibasic heptahydrate in a concentration of about 0.03 g/L.