Polymerized Hemoglobin Media and Its Use in Isolation and Transplantation of Islet Cells

• US 20090004159A1
• Filed: 01/24/2007
• Published: 01/01/2009
• Est. Priority Date: 01/24/2006
• Status: Abandoned Application

• Current Assignee: Board of Trustees of The University of Illinois
• Original Assignee: Board of Trustees of The University of Illinois
• Inventors: Oberholzer, Jose
• Application Number:
  US12/161,542
• Publication Number:
  US 20090004159A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/407 Liver; Hepatocytes
• CPC Class Codes: A61K 38/42 Haemoglobins; Myoglobins
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

Solutions and suspensions comprising polymerized hemoglobin derived from human blood are disclosed. The solutions and suspensions may comprise cell culture medium, an enzyme (such as a protease), and/or a buffer. Processes of preparing the solutions and suspensions are also disclosed. The solutions and suspensions may be employed in methods of isolating mammalian cells, such as pancreatic islets, methods of preserving mammalian tissue and organs, methods of aiding the recovery of mammalian cells following their isolation, methods of maintaining mammalian cells, methods of propagating mammalian cells, and methods of treating a mammal with diabetes.

Claims

1. A cell suspension comprising (a) polymerized hemoglobin derived from mammalian blood and (b) mammalian cells enzymatically produced from a mammalian tissue.

2. The suspension of claim 1, further comprising cell culture medium.

3. The suspension of claim 2, further comprising a protease.

4. The suspension of claim 3, wherein the suspension is oxygenated.

5. The suspension of claim 4, wherein the polymerized hemoglobin is derived from human blood.

6. The suspension of claim 5, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

7. The suspension of claim 1, wherein the mammalian cells comprise pancreatic islets.

8. The suspension of claim 7, further comprising cell culture medium.

9. The suspension of claim 8, further comprising an enzyme.

10. The suspension of claim 9, wherein the suspension is oxygenated.

11. The suspension of claim 10, wherein the polymerized hemoglobin is derived from human blood.

12. The suspension of claim 11, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

13. The suspension of claim 9, wherein the cell culture medium is RPMI and the enzyme is a protease.

14. A method of isolating cells from a tissue in a mammal, comprising contacting the mammal with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) an enzyme for producing cells from said mammalian tissue.

15. The method of claim 14, wherein the tissue is pancreatic tissue.

16. The method of claim 15, wherein the tissue is human pancreatic tissue and the polymerized hemoglobin is derived from human blood.

17. The method of claim 16, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

18. The method of claim 16, wherein the solution is oxygenated.

19. The method of claim 16, wherein the enzyme is a protease.

20. The method of claim 16, wherein said contacting is achieved by intraductal administration.

21. The method of claim 16, further comprising washing the cells produced from human pancreatic tissue with cell culture medium following said contacting to yield pancreatic islets.

22. The method of claim 21, further comprising purifying the pancreatic islets by density gradient centrifugation following said washing.

23. A method of treating a mammal with diabetes by transplanting to the mammal an effective amount of the pancreatic islets isolated and purified according to the method of claim 22.

24. A method of preserving cells enzymatically produced from mammalian tissue, comprising contacting the tissue with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.

25. The method of claim 24 wherein the polymerized hemoglobin is derived from human blood.

26. The method of claim 25, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

27. The method of claim 26, wherein the solution is oxygenated.

28. A method of aiding the recovery of mammalian cells following their isolation from tissue, comprising contacting the cells with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.

29. The method of claim 28 wherein the polymerized hemoglobin is derived from human blood.

30. The method of claim 29, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

31. The method of claim 30, wherein the solution is oxygenated.

32. A method of maintaining mammalian cells isolated from tissue, comprising contacting the cells with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.

33. The method of claim 32 wherein the polymerized hemoglobin is derived from human blood.

34. The method of claim 33, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

35. The method of claim 34, wherein the solution is oxygenated.

36. A method of propagating mammalian cells isolated from tissue, comprising contacting the cells with a solution comprising (a) polymerized hemoglobin derived from mammalian blood and (b) cell culture medium.

37. The method of claim 36 wherein the polymerized hemoglobin is derived from human blood.

38. The method of claim 37, wherein the polymerized hemoglobin derived from human blood is pyridoxylated.

39. The method of claim 38, wherein the solution is oxygenated.

40. A method for treating a mammal with diabetes comprising the step of administering to the mammal an effective amount of a cell suspension comprising polymerized mammalian hemoglobin and pancreatic islet cells.