Methods for rejuvenating red blood cells

1. A method for treating or preventing ischemia in a subject, comprising:

(a) obtaining a volume of blood;

(b) performing a separation on the volume of blood, wherein a volume comprising leukocytes or plasma is separated from red blood cells;

(c) measuring a level of a metabolic indicator in the red blood cells;

(d) rejuvenating the red blood cells with an enhancement composition to form rejuvenated red blood cells;

(e) measuring the level of the metabolic indicator in the rejuvenated red blood cells; and

(f) delivering a volume of rejuvenated blood comprising said rejuvenated red blood cells to the subject in need thereof, wherein the quantity is based on the result of level of the metabolic indicator as measured in step (e), thereby preventing or treating ischemia in said subject.

2. The method according to claim 1 , wherein the enhancement composition comprises:

(i) from about 25 g/L to about 30 g/L of inosine;

(ii) from about 0.2 g/L to about 2 g/L of adenine;

(iii) from about 5 g/L to about 15 g/L of pyruvate; and

(iv) from about 17 g/L to about 23 g/L of sodium phosphate.

3. The method according to claim 1 , wherein steps (b) through (e) are performed within 24 hours of step (a).

4. The method according to claim 1 , wherein the metabolic indicator is 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or both, in the red blood cells.

5. The method according to claim 4 , further comprising measuring a level of the metabolic indicator in the subject prior to administering the rejuvenated red blood cells, and calculating the volume of rejuvenated red blood cells necessary to dose the subject to achieve a level of the metabolic indicator in the subject that is greater than the measured level of the metabolic indicator in the subject.

6. The method according to claim 1 , wherein the red blood cells are recombined with the volume comprising leukocytes or plasma before step (f).

7. The method according to claim 1 , wherein the metabolic indicator is measured by a collecting a sample of blood on a tag, a chip, a glass plate, or a pregnancy test-style strip.

8. The method according to claim 1 , wherein rejuvenating the red blood cells results in red blood cell 2,3-diphosphoglycerate concentration of greater than about 15 μmol/L/gHb.

9. A method of treating ischemia in a subject, comprising:

(a) obtaining a volume of red blood cells;

(b) contacting the red blood cells with an enhancement composition to form rejuvenated red blood cells;

(c) determining a level of 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or both, in the rejuvenated red blood cells;

(d) calculating a transfusion volume of rejuvenated blood comprising said rejuvenated red blood cells necessary to dose the subject to normal or higher concentration of 2,3-DPG, ATP, or both, based on the results efconcentration of 2,3-DPG, ATP, or both, as determined in step (c); and

(e) administering the transfusion volume of the rejuvenated red blood cells to the subject in need thereof, thereby treating ischemia in said subject.

10. The method according to claim 9 , wherein the unit of blood is autologous to the subject.

11. The method according to claim 9 , wherein the enhancement composition comprises:

(i) from about 25 g/L to about 30 g/L of inosine;

(ii) from about 0.2 g/L to about 2 g/L of adenine;

(iii) from about 5 g/L to about 15 g/L of pyruvate; and

(iv) from about 17 g/L to about 23 g/L of sodium phosphate.

12. The method according to claim 9 , wherein determining includes using bioluminescence to quantify the level of 2,3-DPG, ATP, or both.

13. The method according to claim 9 , wherein the subject is in anticipation of surgery.