Methods of using regenerative cells in the treatment of musculoskeletal disorders

• US 7,771,716 B2
• Filed: 07/01/2004
• Issued: 08/10/2010
• Est. Priority Date: 12/07/2001
• Status: Expired due to Fees

• Current Assignee: Lorem Vascular Pte. Ltd.
• Original Assignee: Cytori Therapeutics Incorporated
• Inventors: Riley, Susan Lynn, Fraser, John K., Hedrick, Marc H., Schreiber, Ronda Elizabeth
• Primary Examiner(s): LANKFORD JR, LEON B
• Application Number:
  US10/884,637
• Publication Number:
  US 20050048644A1
• Time in Patent Office: 2,231 Days
• Field of Search: None
• US Class Current: 424/93.7
• 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: A61L 2300/40 characterised by a specific therapeutic activity or mode of action
• CPC Class Codes: A61L 2300/412 Tissue-regenerating or healing or proliferative agents
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 2430/02 for reconstruction of bones; weight-bearing implants
• CPC Class Codes: A61L 2430/06 for cartilage reconstruction, e.g. meniscus
• CPC Class Codes: A61L 2430/40 Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3608 Bone, e.g. demineralised bone matrix [DBM], bone powder
• CPC Class Codes: A61L 27/365 Bones
• CPC Class Codes: A61L 27/3654 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61L 27/3847 Bones
• CPC Class Codes: A61L 27/3852 Cartilage, e.g. meniscus
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/52 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61P 19/02 : for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/04 : for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/08 : for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: C12N 5/0667 : Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

Regenerative cells present in adipose tissue are used to treat patients, including patients with musculoskeletal diseases or disorders. Methods of treating patients include processing adipose tissue to deliver a concentrated amount of regenerative cells obtained from the adipose tissue to a patient. The methods may be practiced in a closed system so that the stem cells are not exposed to an external environment prior to being administered to a patient. Accordingly, in a preferred method, regenerative cells present in adipose tissue are placed directly into a recipient along with such additives necessary to promote, engender or support a therapeutic musculoskeletal benefit.

Claims

1. A method for promoting bone, cartilage, skeletal muscle, tendon, ligament or proteoglycan-rich matrix formation in a subject comprising:

identifying a subject with a musculoskeletal disorder;

providing a programmable, automated adipose-derived stem cell processing unit for concentrating a population of cells comprising adipose-derived stem cells from adipose tissue;

wherein the programmable, automated adipose-derived stem cell processing unit for concentrating the population of cells comprising adipose-derived stem cells comprises;

a tissue collection chamber that is configured to receive unprocessed adipose tissue that is removed from a patient, wherein said tissue collection chamber is defined by a closed system;

a first filter that is disposed within said tissue collection chamber, wherein said first filter is configured to retain a first component of said unprocessed adipose tissue and pass a second component of said unprocessed adipose tissue, such that said first filter substantially separates said first component from said second component, and wherein said first component comprises a cell population that comprises adipose-derived stem cells and said second component comprises lipid, mature adipocytes, and saline;

a processing chamber, which is configured to receive said first component comprising said cell population that comprises adipose-derived stem cells from said tissue collection chamber, wherein said processing chamber is within said closed system;

a conduit configured to allow passage of said first component comprising said cell population comprising adipose-derived stem cells from said tissue collection chamber to said processing chamber while maintaining a closed system;

a cell concentrator disposed within said processing chamber, which is configured to facilitate the concentration of said first component comprising said cell population that comprises adipose-derived stem cells so as to obtain a concentrated population of cells that comprises adipose-derived stem cells, wherein said cell concentrator comprises a centrifugation device or a filter;

an outlet configured to allow the aseptic removal of said concentrated population of cells that comprise adipose-derived stem cells; and

a programmable processing device capable of communicating with and controlling the tissue collection chamber or the processing chamber, or both, and the cell concentrator;

introducing adipose tissue obtained from said subject to said programmable, automated system for concentrating the population of cells comprising adipose-derived stem cells;

processing said adipose tissue obtained from said subject in said programmable, automated system for concentrating the population of cells comprising adipose-derived stem cells so as to obtain a concentrated population of cells that comprises adipose-derived stem cells;

mixing said concentrated population of cells that comprises adipose-derived stem cells in a physiologic solution or a hydrogel liquid; and

introducing the mixture of the concentrated population of cells that comprises adipose-derived stem cells into said subject by injection at a site where bone, cartilage, skeletal muscle, tendon, ligament or proteoglycan-rich matrix formation is desired.

2. The method of claim 1, wherein the cell concentrator comprises a centrifuge.

3. The method of claim 1, wherein the concentrated population of cells that comprises adipose-derived stem cells further comprises progenitor cells.

4. The method of claim 1, wherein the concentrated population of cells that comprises adipose-derived stem cells is suitable for infusion into a patient.

5. The method of claim 1, wherein said site is an intervertebral disc.

6. The method of claim 5, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced by direct injection into the intervertebral disc.

7. The method of claim 5, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced by injection into a gel that is placed within said subject.

8. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of an intervertebral body spinal fusion.

9. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells are introduced into a subject in need of an intertransverse process spinal fusion.

10. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of cartilage formation.

11. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of cartilage formation at the end of long bones.

12. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of cartilage formation in a knee joint.

13. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of cartilage formation in the ribs.

14. The method of claim 10, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced by direct injection into the site of cartilage formation.

15. The method of claim 10, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced by injection into a gel that is placed within said subject.

16. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of repair of a long bone fracture.

17. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of bone or cartilage formation.

18. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of tendon repair.

19. The method of claim 18, wherein the tendon is at a tendon-bone insertion site.

20. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of skeletal muscle formation.

21. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of ligament repair at a ligament-muscle insertion site.

22. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need ligament repair at a ligament-bone insertion site.

23. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is introduced into a subject in need of ligament repair.

24. The method of claim 1, wherein the programmable, automated adipose-derived stem cell processing unit further comprises a washing solution source.

25. The method of claim 24, wherein the washing solution is saline.

26. The method of claim 1, wherein the programmable, automated adipose-derived stem cell processing unit further comprises a disaggregation agent source.

27. The method of claim 26, wherein the disaggregation agent is selected from the group consisting of collagenase, trypsin, lipase, hyaluronidase, deoxyribonuclease, Liberase HI, and pepsin, or mixtures thereof.

28. The method of claim 1, wherein the processing chamber of the programmable, automated adipose-derived stem cell processing unit comprises one or more centrifuge chambers to hold the disaggregated tissue present in the processing chamber during centrifugation of the disaggregated tissue in the centrifuge.

29. The method of claim 28, wherein the one or more centrifuge chambers further comprise one or more output chambers to collect the concentrated population of cells that comprises adipose-derived stem cells during centrifugation of the disaggregated tissue in the centrifugation device.

30. The method of claim 29, wherein the one or more output chambers are tilted at a different angle during centrifugation and a different angle for collection of the concentrated population of cells that comprises adipose-derived stem cells.

31. The method of claim 29, wherein the one or more output chambers can be manually re-positioned to arrive at the angle for collection of the concentrated population of cells that comprises adipose-derived stem cells.

32. The method of claim 31, wherein the programmable, automated system for concentrating adipose-derived stem cells further comprises a syringe.

33. The method of claim 1, wherein the processing chamber of the programmable, automated adipose-derived stem cell processing unit is comprised of a rotating seal to facilitate collection of the disaggregated tissue in a sterile manner.

34. The method of claim 33, wherein the rotating seal is comprised of a rotating shaft, two or more bearings, three or more lip seals, an outer housing, a circular channel and a circular spring.

35. The method of claim 33, wherein the rotating seal is comprised of a rotating shaft, a single rubber seal, an air gasket, one or more spring loaded seals, and two ceramic disks.

36. The method of claim 1, wherein the programmable, automated adipose-derived stem cell processing unit further comprises of a waste chamber.

37. The method of claim 36, wherein the waste chamber receives the non-adipose-derived stem cell components of the tissue removed from said subject.

38. The method of claim 37, wherein the non-adipose-derived stem cell components present in the waste chamber are centrifuged to substantially separate and concentrate collagen, proteins, lipids, adipocytes or matrix components.

39. The method of claim 37, wherein the non-adipose-derived stem cell components present in the waste chamber are filtered to substantially separate and concentrate collagen, proteins, lipids, adipocytes or matrix components.

40. The method of claim 1, wherein the collection chamber and the processing chamber of the programmable, automated adipose-derived stem cell processing unit are connected through tubing.

41. The method of claim 40, wherein the tubing is further comprised of sensors.

42. The method of claim 17, wherein the tissue collection chamber and the processing chamber of the automated system further comprise one or more valves, pumps, or sensors or combinations thereof.

43. The method of claim 40, wherein the tissue collection chamber, the processing chamber or the tubing are disposable.

44. The method of claim 1, wherein the centrifugation device or the processing device of the automated system are re-usable.

45. The method of claim 42, wherein the centrifugation device further comprises a centrifuge motor, a centrifuge motor controller and a centrifuge brake controller.

46. The method of claim 1, wherein the processing device of the programmable, automated adipose-derived stem cell processing unit controls the processing chamber and the centrifuge chamber by directing tissue flow through the use of one or more pumps.

47. The method of claim 46, wherein the pumps are peristaltic pumps.

48. The method of claim 47, wherein the pumps are re-usable.

49. The method of claim 1, wherein the processing device further comprises a user interface for a user to input parameters into the system.

50. The method of claim 1, wherein the programmable processing device further comprises a display screen to display instructions that guide a user to input parameters, confirm pre-programmed steps, warn of errors or combinations thereof.

51. The method of claim 1, wherein a portion of the tissue removed from said subject is used to substantially separate and concentrate said population of cells that comprises adipose-derived stem cells and a remaining portion is set aside.

52. The method of claim 51, wherein the remaining portion of tissue is set aside in a sample chamber.

53. The method of claim 1, wherein one or more additives are added to the concentrated population of cells that comprises adipose-derived stem cells.

54. The method of claim 53, wherein the one or more additives are growth factors, re-suspension fluids, cell culture reagents, cell expansion reagents, cell preservation reagents cell modification reagents, cells, tissue, or tissue fragments or combinations thereof.

55. The method of claim 1, wherein said concentrated population of cells that comprises adipose-derived stem cells is grown in cell culture.

56. The method of claim 55, wherein said concentrated population of cells that comprises adipose-derived stem cells is grown in culture conditions that promote differentiation towards a osteogenic phenotype.

57. The method of claim 1, wherein the concentrated population of cells that comprises adipose-derived stem cells is placed on a surgical device or implant.

58. The method of claim 57, wherein the surgical device is a spinal fusion device.

59. The method of claim 57, wherein the surgical implant is a scaffold.

60. The method of claim 59, wherein the scaffold is porous.

61. The method of claim 1, wherein the concentrated population of cells that comprises adipose-derived stem cells are placed in a gel.

62. The method of claim 1, the programmable, automated adipose-derived stem cell processing unit, is connected to a second system that comprises a means for culturing said concentrated population of cells that comprises adipose-derived stem cells.

63. A method for promoting bone, cartilage, skeletal muscle, tendon, ligament or proteoglycan-rich matrix formation in a subject comprising:

identifying a subject with a musculoskeletal disorder;

providing a programmable, automated adipose-derived stem cell processing unit for concentrating a population of cells comprising adipose-derived stem cells from adipose tissue;

wherein the programmable, automated adipose-derived stem cell processing unit for concentrating the population of cells comprising adipose-derived stem cells comprises;

a tissue collection chamber that is configured to receive unprocessed adipose tissue that is removed from a patient, wherein said tissue collection chamber is defined by a closed system;

a first filter that is disposed within said tissue collection chamber, wherein said first filter is configured to retain a first component of said unprocessed adipose tissue and pass a second component of said unprocessed adipose tissue, such that said first filter substantially separates said first component from said second component, and wherein said first component comprises a cell population that comprises adipose-derived stem cells and said second component comprises lipid and mature adipocytes;

a processing chamber, which is configured to receive said first component comprising said cell population that comprises adipose-derived stem cells from said tissue collection chamber, wherein said processing chamber is within said closed system;

a conduit configured to allow passage of said first component comprising said cell population comprising adipose-derived stem cells from said tissue collection chamber to said processing chamber while maintaining a closed system;

a cell concentrator disposed within said processing chamber, which is configured to facilitate the concentration of said first component comprising said cell population that comprises adipose-derived stem cells so as to obtain a concentrated population of cells that comprises adipose-derived stem cells, wherein said cell concentrator comprises a centrifugation device or a filter;

an outlet configured to allow the aseptic removal of said concentrated population of cells that comprise adipose-derived stem cells; and

a programmable processing device capable of communicating with and controlling the tissue collection chamber or the processing chamber, or both, and the cell concentrator;

introducing adipose tissue obtained from said subject to said programmable, automated system for concentrating the population of cells comprising adipose-derived stem cells;

processing said adipose tissue obtained from said subject in said programmable, automated system for concentrating the population of cells comprising adipose-derived stem cells so as to obtain a concentrated population of cells that comprises adipose-derived stem cells;

mixing said concentrated population of cells that comprises adipose-derived stem cells in a physiologic solution or a hydrogel liquid; and

introducing the mixture of the concentrated population of cells that comprises adipose-derived stem cells into said subject by injection at a site where bone, cartilage, skeletal muscle, tendon, ligament or proteoglycan-rich matrix formation is desired.