Methods for rejuvenating red blood cells

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

(a) obtaining a volume of fresh blood from the subject;

(b) performing a separation on the fresh blood, wherein plasma and leukocytes are delivered to the subject, and fresh red blood cells are isolated;

(c) rejuvenating the fresh red blood cells with an enhancement composition to form rejuvenated fresh red blood cells, wherein the rejuvenated fresh red blood cells have adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) levels equal to or greater than ATP and 2,3-DPG levels normally found in the subject's blood; and

(d) delivering the rejuvenated fresh red blood cells to the subject, wherein the performing a separation, rejuvenating and delivering are performed within about 24 hours of the time of obtaining the fresh blood from the 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 , further comprising performing a diagnostic test on the rejuvenated red blood cells.

4. The method according to claim 3 , wherein the diagnostic test measures the concentrations of 2,3-diphosphoglycerate (2,3-DPG) or adenine triphosphate (ATP) in the rejuvenated red blood cells.

5. The method according to claim 4 , further comprising calculating the volume of rejuvenated blood necessary to dose the subject to obtain normal or higher levels of 2,3-DPG and/or ATP in the subject.

6. The method according to claim 3 , wherein the diagnostic test is performed by a device physically attached to the unit of blood.

7. The method according to claim 3 , wherein the diagnostic test comprises 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 the method is performed in anticipation of surgery on the subject.

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

10. A method of enhancing the ability of blood to oxygenate tissue in a subject in need thereof comprising:

(a) obtaining a unit of fresh blood;

(b) performing a first diagnostic test on the fresh blood to determine the concentration of a marker selected from 2,3-diphosphoglycerate (2,3-DPG), adenine triphosphate (ATP), or combinations thereof;

(c) contacting the fresh blood with an enhancement composition to form rejuvenated fresh blood;

(d) performing a second diagnostic test on the rejuvenated fresh blood to determine the concentration of the marker;

(e) comparing the concentration of the marker from the first diagnostic test to the concentration of the marker from the second diagnostic test and determining a volume of rejuvenated fresh blood necessary to dose the subject to normal or higher levels of 2,3-DPG and/or ATP in the subject's systemic circulation; and

(f) administering the volume of rejuvenated fresh blood, determined in step (e) to the subject;

wherein the obtaining, performing a first diagnostic test, contacting, performing a second diagnostic test, comparing and administering are performed within about 24 hours of the time of obtaining the fresh blood from the subject.

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

12. The method according to claim 10 , 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.

13. The method according to claim 12 , wherein the diagnostic test is performed on a chip.

14. The method according to claim 12 , wherein the subject is in anticipation of surgery.

15. A method of performing a transfusion comprising:

(a) obtaining a unit of fresh blood;

(b) performing a diagnostic test on the unit of fresh blood to determine 2,3-diphosphoglycerate (2,3-DPG) concentration in the blood;

(c) rejuvenating the fresh blood if the 2,3-DPG concentration is lower than about 10 μmol/L/gHb to form rejuvenated fresh blood, wherein the rejuvenating is performed with an enhancement composition comprising, inosine, adenine, pyruvate, and sodium phosphate; and

(d) delivering the rejuvenated fresh blood to a subject in need thereof;

wherein the obtaining, performing a diagnostic test, rejuvenating and delivering are performed within about 24 hours of the time of obtaining the fresh blood from the subject.

16. A method for treating or preventing ischemia in a human or other mammalian subject, comprising:

(a) obtaining a volume of fresh blood from the subject;

(b) performing a separation on the fresh blood, wherein plasma and leukocytes are delivered to the subject, and fresh red blood cells are isolated;

(c) rejuvenating the fresh red blood cells with an enhancement composition to form rejuvenated fresh red blood cells, wherein the concentration of 2,3-diphosphoglycerate (2,3-DPG) concentration in the rejuvenated red blood cells is greater than about 15 μmol/L/gHb; and

(d) delivering the rejuvenated fresh red blood cells to the subject;

wherein the obtaining, performing a separation, rejuvenating, and administering are performed within about 24 hours of the time of obtaining the fresh blood from the subject.

17. The method according to claim 16 , wherein the delivering of the rejuvenated fresh red blood cells is performed within 6 hours after the obtaining of the volume of fresh blood from the subject.

18. The method according to claim 16 , 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.

19. The method according to claim 16 , further comprising performing a diagnostic test on the rejuvenated fresh red blood cells, measuring the concentrations of 2,3-diphosphoglycerate (2,3-DPG) or adenine triphosphate (ATP) in the rejuvenated red blood cells.

20. The method according to claim 16 , wherein the method is performed in anticipation of surgery on the subject.