Lyophilized and reconstituted red blood cell compositions

• US 5,178,884 A
• Filed: 06/29/1989
• Issued: 01/12/1993
• Est. Priority Date: 05/18/1988
• Status: Expired due to Fees

• Current Assignee: Cobe Cardiovascular Incorporated (LivaNova Plc)
• Original Assignee: Cryopharm Corp.
• Inventors: Goodrich, Raymond P., Hackett, Roger W., Williams, Christine M.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/374,171
• Time in Patent Office: 1,293 Days
• Field of Search: 435/2, 424/529, 424/533, 514/6
• US Class Current: 424/533
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0226 Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
• CPC Class Codes: A61K 35/14 Blood; Artificial blood perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42
• CPC Class Codes: A61K 35/18 Erythrocytes haemoglobin A61K38/42

Abstract

A process and medium are disclosed for the lyophilization of red blood cells which comprises the use of solutions including monosaccharide hexoses and pentoses, and biocompatible amphipathic polymers to permit the reconstitution of viable red blood cells. Also disclosed are lyophilized and reconstituted erythrocyte and hemosome containing compositions.

Claims

1. A transfusably-useful erythrocyte-containing and/or hemosome-containing composition reconstituted after lyophilization characterized by a P₅₀ in the range of about 26-28, an oxyhemoglobin content greater than about 90% of that of normal red blood cells, morphology,and normal ATP metabolism.

2. A composition according to claim 1 comprising erythrocytes having discocytic or stomatocytic morphology.

3. A composition according to claim 2 further comprising an additive comprising a polymer or mixture of polymers having a molecular weight of from about 1 K to about 360 K in a concentration of about 12 to 30% by weight.

4. A composition according to claim 3 wherein said polymers are amphipathic.

5. A composition according to claim 3 wherein said polymers have molecular weights in the range of about 2.5 K to 360 K.

6. A composition according to claim 3, 4 or 5 wherein said polymer is selected from the group consisting of polyvinylpyrrolidone, hydroxyethyl starch and dextran.

7. A composition according to claim 6 wherein said polymer comprises polyvinylpyrrolidone of average molecular weight of 40 K.

8. A composition according to claim 7 wherein said polyvinylpyrrolidone is present in said solution in a final concentration of about 14.4% by weight.

9. A composition according to claim 8 wherein said solution further comprises a monosaccharide in a final concentration of about 7.0 to 37.5% by weight.

10. A composition according to claim 2 wherein said composition further comprises a polyanion selected from the group consisting of inositol hexaphosphate, adenosine triphosphate, pyrophosphate and tripolyphosphate.

11. A composition according to claim 10 wherein said polyanion comprises inositol hexaphosphate.

12. A composition according to claim 2 further comprising polyethylene glycol of a molecular weight in a range of about 3 to 5 KD.

13. A lyophilized erythrocyte-containing and/or hemosome-containing composition characterized by a phosphorous-31 NMR lineshape characteristic of a bilayer in a gel phase.

14. A composition according to claim 13 comprising erythrocytes.

15. A composition according to claim 14 further comprising an additive comprising a polymer or mixture of polymers having a molecular weight of from about 1 K to about 360 K in a concentration of about 12 to 30% by weight.

16. A composition according to claim 15 wherein said polymers are amphipathic.

17. A composition according to claim 15 wherein said polymers have molecular weights in the range of about 2.5 K to 360 K.

18. A composition according to claim 15, 16 or 17 wherein said polymer is selected from the group consisting of polyvinylpyrrolidone, hydroxyethyl starch and dextran.

19. A composition according to claim 18 wherein said polymer comprises polyvinylpyrrolidone of average molecular weight of 40 K.

20. A composition according to claim 14 wherein said solution further comprises a monosaccharide.

21. A composition according to claim 15, 16 or 17 further characterized by a phosphorous-31 NMR spectrum linewidth reduced by at least 8% as compared to the linewidth of a corresponding erythrocyte-containing composition lyophilized without said additive.