Rapid infusion of autologous bone marrow derived stem cells

• US 9,439,930 B2
• Filed: 06/04/2014
• Issued: 09/13/2016
• Est. Priority Date: 01/12/2013
• Status: Active Grant

• Current Assignee: The Thermogenesis Corporation (Cesca Therapeutics Incorporated)
• Original Assignee: Cesca Therapeutics Incorporated
• Inventors: Ponemone, Venkatesh, Harris, Kenneth, Sethi, Dalip, Cohan, Gary R.
• Primary Examiner(s): FOX, ALLISON M
• Application Number:
  US14/296,360
• Publication Number:
  US 20140341861A1
• Time in Patent Office: 832 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 composition comprising:

(a) a first cell population that comprises bone marrow stem cells and/or progenitor cells,

(b) an anticoagulant,

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

(d) red blood cells,

wherein the composition has a viscosity of 1.5-3.5 centipoise (cP) measured at 37°

C.

2. The composition of claim 1, wherein the first cell population that comprises bone marrow stem cells and/or progenitor cells is autologous.

3. The composition of claim 2, wherein the first cell population that comprises bone marrow stem cells and/or progenitor cells is from a medullary space within a bone of a patient.

4. The composition of claim 1, wherein said anticoagulant is selected from the group consisting of a coumarin, a vitamin K antagonist, an indirect thrombin inhibitor, heparin, a factor Xa inhibitor, a direct thrombin inhibitor, batroxobin, hemetin, a purified plant extract, EDTA, citrate, oxalate, and a nitrophorin.

5. The composition of claim 1, wherein said anticoagulant comprises bivalirudin.

6. The composition of claim 5, wherein said bivalirudin is present in said composition at a concentration of 1 mg/mL-3 mg/mL.

7. The composition of claim 5, wherein said bivalirudin is present in said composition at a concentration of 1.5 mg/mL-2.5 mg/mL.

8. The composition of claim 5, wherein said bivalirudin is present in said composition at a concentration of 1.8 mg/mL-2.2 mg/mL.

9. The composition of claim 5, wherein said bivalirudin is present in said composition at a concentration of 2.0 mg/mL.

10. The composition of claim 1, wherein said viscosity measured at 37°

C. is 1.8 cP-3.0 cP.

11. The composition of claim 1, wherein said viscosity measured at 37°

C. is 2.0 cP-2.5 cP.

12. The composition of claim 1, wherein said viscosity measured at 37°

C. is 3.1 cP-3.5.

13. The composition of claim 1, wherein said aqueous buffer comprises an ionic salt.

14. The composition of claim 1, wherein said aqueous buffer comprises sodium chloride and the amount of said sodium chloride in said composition is 0.9%.

15. The composition of claim 1, wherein said cell population that comprises bone marrow cells comprises mononuclear cells that comprise at least a stem cell and/or progenitor cell selected from the group consisting of hematopoietic stem cells, mesenchymal stem cells, endothelial progenitor cells, and CXCR4 positive cells.

16. The composition of claim 1, wherein said cell population that comprises bone marrow cells comprises at least 10⁷ mononuclear cells, not more than 10⁹ mononuclear cells, at least 10⁴ hematopoietic stem cells, at least 5×

10² mesenchymal stem cells, at least 5×

10² endothelial progenitor cells, and/or at least 5×

10³ CXCR4 positive cells.

17. The composition of claim 1, wherein said composition demonstrates an apoptosis or other cell death rate of not more than about 40% one hour after delivery at a flow rate of 2.5 mL/min, through a catheter having a lumen size of approximately 0.36 mm and using a 5 mL syringe holding pressure to 28.39 psi with a plunger force of 4.85 lbf.

18. The composition of claim 1, wherein said composition demonstrates an apoptosis or other cell death rate of not more than about 40% one hour after delivery through a catheter if subjected to a maximum shear of not more than 9101/second.

19. A method of making a regenerative cell composition comprising:

(a) providing a first cell population that comprises bone marrow stem cells and/or progenitor cells;

(b) mixing an anticoagulant with said first cell population so as to produce a composition comprising said first cell population and the anticoagulant;

(c) enriching bone marrow stem cells within said composition comprising said cell population and the anticoagulant so as to produce a second cell population that comprises enriched bone marrow stem cells and/or progenitor cells;

(d) isolating a fraction comprising said second cell population that comprises enriched bone marrow stem cells and/or progenitor cells; and

(e) adjusting the viscosity of said fraction using red blood cells to a viscosity of 1.5 cP-5.0 cP measured at 37°

C. so as to make said regenerative cell composition.

20. The method of claim 19, wherein said anticoagulant comprises bivalirudin.

21. The method of claim 19, comprising adjusting the viscosity of said fraction using red blood to a viscosity of 1.5 cP-3.0 cP measured at 37°

C.

22. The method of claim 19, comprising adjusting the viscosity of said fraction using red blood to a viscosity of 2.0 cP-2.5 cP measured at 37°

C.

23. The method of claim 19, comprising adjusting the viscosity of said fraction using red blood to a viscosity of 3.1 cP-5.0 cP measured at 37°

C.

24. A regenerative cell composition made by a process comprising:

(a) providing a first cell population that comprises bone marrow stem cells and/or progenitor cells;

(b) mixing an anticoagulant with said first cell population so as to produce a composition comprising said first cell population and the anticoagulant;

(c) enriching bone marrow stem cells within said composition comprising said cell population and the anticoagulant so as to produce a second cell population that comprises enriched bone marrow stem cells and/or progenitor cells;

(d) isolating a fraction comprising said second cell population that comprises enriched bone marrow stem cells and/or progenitor cells; and

(e) adjusting the viscosity of said fraction using red blood cells to a viscosity of 1.5 cP-5.0 cP measured at 37°

C. so as to make said regenerative cell composition.

25. The regenerative cell composition of claim 24, wherein the process comprises adjusting the viscosity of said fraction using red blood to a viscosity of 1.5 cP-3.0 cP measured at 37°

C.

26. The regenerative cell composition of claim 24, wherein the process comprises adjusting the viscosity of said fraction using red blood to a viscosity of 2.0 cP-2.5 cP measured at 37°

C.

27. The regenerative cell composition of claim 24, wherein the process comprises adjusting the viscosity of said fraction using red blood to a viscosity of 3.1 cP-5.0 cP measured at 37°

C.

28. The regenerative cell composition of claim 24, wherein said second cell population comprises mononuclear cells that comprise at least a stem cell and/or progenitor cell selected from the group consisting of hematopoietic stem cells, mesenchymal stem cells, endothelial progenitor cells, and CXCR4 positive cells.

29. The regenerative cell composition of claim 24, wherein said second cell population comprises at least 10⁷ mononuclear cells, not more than 10⁹ mononuclear cells, at least 10⁴ hematopoietic stem cells, at least 5×

10² mesenchymal stem cells, at least 5×

10² endothelial progenitor cells, and/or at least 5×

10³ CXCR4 positive cells.

30. The regenerative cell composition of claim 24, wherein said second cell population demonstrates an apoptosis or other cell death rate of not more than about 40% one hour after delivery at a flow rate of 2.5 mL/min, through a catheter having a lumen size of approximately 0.36 mm and using a 5 mL syringe holding pressure to 28.39 psi with a plunger force of 4.85 lbf.

31. The regenerative cell composition of claim 24, wherein said second cell population demonstrates an apoptosis or other cell death rate of not more than about 40% one hour after delivery through a catheter if subjected to a maximum shear of not more than 9101/second.