Rapid infusion of autologous bone marrow derived stem cells

• US 9,393,269 B2
• Filed: 06/05/2014
• Issued: 07/19/2016
• Est. Priority Date: 01/12/2013
• Status: Active Grant

• Current Assignee: The Thermogenesis Corporation (Cesca Therapeutics Incorporated)
• Original Assignee: Cesca Therapeutics Incorporated
• Inventors: Sethi, Dalip, Ponemone, Venkatesh, Harris, Kenneth, Cohan, Gary R.
• Primary Examiner(s): FOX, ALLISON M
• Application Number:
  US14/297,557
• Publication Number:
  US 20140286869A1
• Time in Patent Office: 775 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61B 5/14503 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors A61B5/1459, A61B5/1464, A61B5/1473, A61B5/1482, A61B5/14865 take precedence
• 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 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61K 38/55 Protease inhibitors
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 49/0004 Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Abstract

The present invention relates to a method and composition for the aspiration, processing, testing and infusion of bone marrow derived stem cells, as an adjuvant treatment in cardiovascular disorders. More specifically, the invention provides for the methods and compositions for the aspiration, analysis, processing, infusate preparation and infusion of bone-marrow derived stem cells, particularly in a rapid point-of-care environment, wherein a centrifugal fractionation and optically monitored separation of the bone marrow yield desired cellular product in the desired concentration and viscosity.

Claims

1. A point-of-care method of ameliorating critical limb ischemia or a condition associated with critical limb ischemia in a subject comprising:

identifying a subject having a critical limb ischemia or a condition associated with critical limb ischemia;

providing to said subject a composition comprising;

(a) a first cell population that comprises CD34⁺ and CD34⁻

bone marrow stem cells,

(b) an anticoagulant, and

(c) an aqueous buffer and/or an autologous serum and/or an autologous plasma fraction,

wherein the composition has a viscosity of 1.5 to 5.0 centipoise (cP) measured at 37°

C. and wherein said composition is provided to said subject at a rate of 1.0 mL per minute.

2. The method of claim 1, wherein the composition is provided to said subject not more than 2 hours after said first cell population is obtained.

3. The method of claim 1, wherein said composition is provided to said subject not more than 90 minutes after said first cell population is obtained.

4. The method of claim 1, wherein said composition is provided to said subject not more than 60 minutes after said first cell population is obtained.

5. The method of claim 1, wherein the blood flow at an ischemic tissue of said subject is blocked for 1-3 minutes or 2 minutes.

6. The method of claim 5, wherein said blood flow is blocked by an inflatable balloon, wherein said balloon is inflated to block at least 80% of blood flow distal to said balloon.

7. The method of claim 1, wherein said composition provided to said subject is not subjected to a maximum shear of more than 9101/second.

8. The method of claim 1, wherein said composition is provided to said subject through a catheter having a lumen size of approximately 0.36 mm.

9. The method of claim 1, wherein said composition is provided intravascularly or intra-arterially.

10. The method of claim 1, further comprising measuring in said subject an increased blood flow, an increase in angiogenesis, an increase in vascularity, or an increase in amputation-free survival rate.

11. The method of claim 1, wherein said composition is provided to said subject at a pressure of 15-60 psi.

12. The method of claim 1, wherein said composition has a viscosity of 1.8 to 5.0 centipoise (cP) measured at 37°

C.

13. The method of claim 1, wherein said CD34⁻

bone marrow stern cells comprise mesenchymal stem cells or stromal cells.

14. The method of claim 13, wherein said mesenchymal stem cells or stromal cells are lineage negative/dim, CD45 negative/dim and CD73 positive cells.

15. The method of claim 1, wherein said first cell population comprises hematopoietic stern cells, mesenchymal stem cells, stromal cells, endothelial progenitor cells, CXCR4 positive cells, or a combination thereof.

16. The method of claim 1, wherein said composition comprises red blood cells.

17. A point-of-care method of ameliorating critical limb ischemia or a condition associated with critical limb ischemia in a subject comprising:

identifying a subject having a critical limb ischemia or a condition associated with critical limb ischemia;

introducing bone marrow aspirate from said subject into a bone marrow processing system;

isolating a stem cell fraction, a plasma fraction, and a red blood cell fraction from said disposable bone marrow processing device;

adding plasma from said plasma fraction and red blood cells from said red blood cell fraction to said stem cell fraction to obtain a composition with a viscosity of 1.5 to 5.0 centipoise(cP) measured at 37°

C.; and

providing said composition with a viscosity of 1.5 to 5.0 centipoise (cP) measured at 37°

C. to said subject.

18. The method of claim 17, wherein said composition comprises an anticoagulant.

19. The method of claim 17, wherein said stem cell fraction comprises CD34⁺ and CD34⁻

bone marrow stem cells.

20. The method of claim 19, wherein said CD34⁻

bone marrow stem cells comprise mesenchymal stem cells or stromal cells.

21. The method of claim 20, wherein said mesenchymal stem cells or stromal cells are lineage negative/dim, CD45 negative/dim and CD73 positive cells.

22. The method of claim 17, wherein said composition has a viscosity of 1.8 to 5.0 centipoise (cP) measured at 37°

C.

23. The method of claim 17, wherein said composition is provided to said subject at urate of 1.0 to 2.0 mL per minute.

24. The method of claim 17, wherein said composition is provided to said subject at a rate of 1.0 mL per minute.

25. The method of claim 17, wherein said composition is provided to said subject not more than 2 hours after said first cell population is obtained.

26. The method of claim 17, wherein said composition is provided to said subject not more than 90 minutes after said first cell population is obtained.

27. The method of claim 17, wherein said composition is provided to said subject not more than 60 minutes after said first cell population is obtained.

28. The method of claim 17, wherein the blood flow at an ischemic tissue of said subject is blocked by an inflatable balloon, wherein said balloon is inflated to block at least 80% of blood flow distal to said balloon.

29. The method of claim 17, wherein said composition provided to said subject is not subjected to a maximum shear of more than 9101/second.

30. The method of claim 17, wherein said composition is provided to said subject through a catheter having a lumen size of approximately 0.36 mm.

31. The method of claim 17, wherein said composition is provided intravascularly or intra-arterially.

32. The method of claim 17, further comprising measuring in said subject an increased blood flow, an increase in angiogenesis, an increase in vascularity, or an increase in amputation-free survival rate.

33. A point-of-care method of ameliorating critical limb ischemia or a condition associated with critical limb ischemia in a subject comprising:

identifying a subject having a critical limb ischemia or a condition associated with critical limb ischemia;

introducing bone marrow aspirate from said subject into a bone marrow processing system that comprises;

(a) a sterile, disposable bone marrow processing device,

(b) a control module comprising a gravitational sensor and an infra-red optical sensor coupled with a microprocessor and motor, which are configured to control the flow, movement, and compartmentalization of separated fractions of bone marrow aspirate,

(c) a docking station that transfers processing data from the control module to a computer, and

(d) a centrifuge;

isolating a stem cell fraction, a plasma fraction, and a red blood cell fraction from said disposable bone marrow processing device;

adding plasma from said plasma fraction and red blood cells from said red blood cell fraction to said stem cell fraction to obtain a composition with a viscosity of 1.5 to 5.0 centipoise (cP) measured at 37°

C; and

providing said composition with a viscosity of 1.5 to 5.0 centipoise (cP) measured at 37°

C. to said subject.

34. The method of claim 33, wherein said composition comprises an anticoagulant.

35. The method of claim 33, wherein said stem cell fraction comprises CD34⁺ bone marrow stem cells and mesenchymal stem cells.

36. The method of claim 33, wherein said stem cell fraction comprises stromal cells.

37. The method of claim 36, wherein said stromal cells are lineage negative/dim, CD45 negative/dim and CD73 positive cells.

38. The method of claim 33, wherein said composition has a viscosity of 1.8 to 5.0 centipoise (cP) measured at 37°

C.

39. The method of claim 33, wherein said composition is provided to said subject at a rate of 1.0 to 2.0 mL per minute.

40. The method of claim 33, wherein said composition is provided to said subject at a rate of 1.0 mL per minute.

41. The method of claim 33, wherein said composition is provided to said subject not more than 2 hours after said first cell population is obtained.

42. The method of claim 33, wherein said composition is provided to said subject not more than 90 minutes after said first cell population is obtained.

43. The method of claim 33, wherein said composition is provided to said subject not more than 60 minutes after said first cell population is obtained.

44. The method of claim 33, wherein the blood flow at an ischemic tissue of said subject is blocked by an inflatable balloon, wherein said balloon is inflated to block at least 80% of blood flow distal to said balloon.

45. The method of claim 33, wherein said composition provided to said subject is not subjected to a maximum shear of more than 9101/second.

46. The method of claim 33, wherein said composition is provided to said subject through a catheter having a lumen size of approximately 0.36 mm.

47. The method of claim 33, wherein said composition is provided intravascularly or intra-arterially.

48. The method of claim 33, further comprising measuring in said subject an increased blood flow, an increase in angiogenesis, an increase in vascularity, or an increase in amputation-free survival rate.

49. The method of claim 33, wherein said composition is provided to said subject at a pressure of 15-60 psi.