Therapeutic platelets and methods
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.