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

• US 20110142808A1
• Filed: 01/28/2011
• Published: 06/16/2011
• 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, José
• Application Number:
  US13/016,758
• Publication Number:
  US 20110142808A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• 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 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 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/42 Haemoglobins; Myoglobins
• CPC Class Codes: A61K 38/482 Serine endopeptidases (3.4.21)
• CPC Class Codes: A61K 38/4886 Metalloendopeptidases (3.4.24), e.g. collagenase
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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

1-13. -13. (canceled)

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