Preparation of a cell concentrate from a physiological solution
The present invention is directed to methods and compositions regarding the preparation of an cell concentrate, such as, for example, an osteogenic cell concentrate, from a physiological solution, such as bone marrow aspirate, blood, or a mixture thereof. In specific embodiments, the invention provides methods and compositions utilizing two physiological solution-processing techniques, particularly in a point of care environment, wherein centrifugation is not employed.
1. A method of preparing a cell concentrate, comprising the steps of:
providing a physiological fluid not previously subjected to centrifugation;
subjecting said physiological fluid to a filter to produce a filter retentate and a permeate fluid, wherein said filter retentate comprises a cell concentrate including both platelets and nucleated cells per unit volume greater than in the physiological fluid, wherein the nucleated cells comprise leukocytes, and wherein said permeate fluid comprises plasma and red blood cells, and wherein the filter comprises a leukocyte reduction filter;
removing the cell concentrate from the filter; and
delivering the cell concentrate to a bone defect in an individual.
2. The method of claim 1 , wherein the physiological fluid comprises bone marrow aspirate, blood, or a mixture thereof.
3. The method of claim 1 , wherein said nucleated cells comprise leukocytes, stem cells, connective tissue progenitor cells, osteoprogenitor cells, chondroprogenitor cells, or a mixture thereof.
4. The method of claim 3 , wherein the stem cells are mesenchymal stem cells, hematopoietic stem cells, or both.
5. The method of claim 3 , wherein the providing step comprises combining an additional fluid with the physiological fluid.
6. The method of claim 5 , wherein the additional fluid is water.
7. The method of claim 5 , wherein the additional fluid is a hypotonic solution comprising sodium chloride.
8. The method of claim 1 , wherein the filter retentate that is delivered to the bone defect in the individual further comprises a scaffold material.
9. The method of claim 8 , wherein the scaffold material is comprised of a block, paste, dust, cement, powder, granule, putty, liquid, gel, solid, or a mixture thereof.
10. The method of claim 8 , wherein the scaffold material is comprised of a ceramic, a polymer, a metal, allograft bone, autograft bone, demineralized bone matrix, or a mixture thereof.
11. The method of claim 8 , wherein the scaffold material is biodegradable.
12. The method of claim 8 , wherein the scaffold material is osteoconductive, osteogenic, osteoinductive, or a combination thereof.
13. The method of claim 8 , wherein the scaffold material is comprised of synthetic material, natural material, or a combination thereof.
14. The method of claim 1 , said method further comprising the step of admixing a biological agent with the filter retentate.
15. The method of claim 8 , said method further comprising the step of admixing a biological agent with the scaffold.
16. The method of claim 15 , wherein the biological agent admixed with the scaffold material is further defined as the biological agent being comprised on the scaffold material, in the scaffold material, or both.
17. The method of claim 8 , further comprising the step of admixing a clotting initiator with the filter retentate, the scaffold material, or both.
18. A method of preparing a a scaffold material/filter retentate mixture, comprising the steps of:
providing a physiological fluid not previously subjected to centrifugation; subjecting said physiological fluid to a filter to produce a filter retentate and a permeate fluid, wherein said filter retentate comprises platelets and nucleated cells per unit volume greater than in the physiological fluid wherein the nucleated cells comprise leukocyes and wherein said permeate fluid comprises plasma and red blood cells, and wherein the filter comprises a leukocyte reduction filter;
removing the filter retentate from the filter; and
admixing a scaffold material to the filter retentate removed from the filter to produce a scaffold material/filter retentate mixture.
19. The method of claim 18 , further comprising the step of delivering the scaffold material/filter retentate mixture to a bone defect in an individual.
20. The method of claim 18 , wherein the physiological fluid comprises bone marrow aspirate, blood, or a mixture thereof.
21. The method of claim 18 , wherein said nucleated cells comprise leukocytes, stem cells, connective tissue progenitor cells, osteoprogenitor cells, chondroprogenitor cells, or a mixture thereof.
22. The method of claim 21 , wherein the stem cells are mesenchymal stem cells, hematopoietic stem cells, or both.
23. The method of claim 18 , wherein the providing step comprises combining an additional fluid with the physiological fluid.
24. The method of claim 23 , wherein the additional fluid is water.
25. The method of claim 23 , wherein the additional fluid is a hypotonic solution.
26. The method of claim 23 , wherein the additional fluid is a hypotonic solution comprising sodium chloride.
27. The method of claim 18 , wherein the scaffold material is comprised of a block, paste, dust, cement, powder, granule, putty, liquid, gel, solid, or a mixture thereof.
28. The method of claim 18 , wherein the scaffold material is comprised of a ceramic, a polymer, a metal, allograft bone, autograft bone, demineralized bone matrix, or a mixture thereof.
29. The method of claim 18 , wherein the scaffold material is biodegradable.
30. The method of claim 18 , wherein the scaffold material is osteoconductive, osteogenic, osteoinductive, or a combination thereof.
31. The method of claim 18 , wherein the scaffold material is comprised of synthetic material, natural material, or a combination thereof.
32. The method of claim 18 , said method further comprising the step of admixing a biological agent with the filter retentate.
33. The method of claim 18 , said method further comprising the step of admixing a biological agent with the scaffold.
34. The method of claim 33 , wherein the biological agent admixed with the scaffold material is further defined as the biological agent being comprised on the scaffold material, in the scaffold material, or both.
35. The method of claim 18 , further comprising the step of admixing a clotting initiator with the filter retentate, the scaffold material, or both.
36. The method of claim 18 , wherein the filter retentate removed from the filter is subjected to at least one further processing step.