Therapeutic platelets and method

Abstract

A dehydrated composition is provided that includes freeze-dried platelets. The platelets are loaded with trehalose which preserves biological properties during freeze-drying and rehydration. The trehalose loading is conducted at a temperature of from greater than about 25° C. to less than about 40° C., most preferably at 37° C., with the loading solution having trehalose in an amount from about 10 mM to about 50 mM. These freeze-dried platelets are substantially shelf-stable and are rehydratable so as to have a normal response to an agonist, for example, thrombin, with virtually all of the platelets participating in clot formation within about three minutes at 37° C.

1 - 32 . (canceled)  
     
     
         33 . A process for increasing the loading efficiency of trehalose into platelets comprising: 
 providing platelets having a first phase transition temperature range and a second phase transition temperature range which is greater than the first phase transition temperature;    disposing the platelets into a trehalose solution to commence loading trehalose into the platelets by uptaking trehalose from the trehalose solution by fluid phase endocytosis; and    heating the trehalose solution to the second phase transition temperature range to increase the loading efficiency of trehalose into the platelets.    
     
     
         34 . The process of  claim 33  wherein said platelets comprise human platelets.  
     
     
         35 . The process of  claim 33  wherein said second phase transition temperature range is greater than about 25° C.  
     
     
         36 . The process of  claim 34  wherein said second phase transition temperature range is greater than about 25° C.  
     
     
         37 . The process of  claim 35  wherein said second phase transition temperature ranges from a temperature greater than about 25° C. to a temperature less than about 40° C.  
     
     
         38 . The process of  claim 34  wherein said second phase transition temperature ranges from a temperature greater than about 25° C. to a temperature less than about 40° C.  
     
     
         39 . The process of  claim 37  wherein said temperature ranges from about 30° C. to less than about 40° C.  
     
     
         40 . The process of  claim 38  wherein said temperature ranges from about 30° C. to less than about 40° C.  
     
     
         41 . A process of preparing loaded platelets comprising: 
 providing platelets selected from a mammalian species; and    loading an oligosaccharide by fluid phase endocytosis into the platelets at a temperature greater than about 25° C. to produce loaded platelets.    
     
     
         42 . The process of  claim 41  wherein said loading comprises loading with an oligosaccharide solution.  
     
     
         43 . The process of  claim 42  wherein said fluid phase endocytosis comprises uptaking external oligosaccharide from the oligosaccharide solution at the temperature greater than about 25° C.  
     
     
         44 . The process of  claim 42  wherein said loading comprises incubating the platelets at the temperature greater than about 25° C. with the oligosaccharide solution.  
     
     
         45 . The process of  claim 41  wherein said loading is without a fixative.  
     
     
         46 . The process of  claim 41  wherein said oligosaccharide comprises trehalose.  
     
     
         47 . The process of  claim 41  wherein said loading of the oligosaccharide into the platelets is at a temperature ranging from greater than about 25° C. to less than about 40° C.  
     
     
         48 . The process of  claim 47  wherein said temperature ranges from about 30° C. to less than about 40° C.  
     
     
         49 . The process of  claim 47  wherein said temperature is about 37° C.  
     
     
         50 . The process of  claim 41  wherein said platelets comprise human platelets.  
     
     
         51 . Loaded platelets produced in accordance with the process of  claim 41 .  
     
     
         52 . Loaded platelets produced in accordance with the process of  claim 33.