Adipose cells for chondrocyte applications

• US 10,206,954 B2
• Filed: 06/13/2014
• Issued: 02/19/2019
• Est. Priority Date: 06/19/2013
• Status: Active Grant

• Current Assignee: Spinalcyte LLC
• Original Assignee: Spinalcyte LLC
• Inventors: O'Heeron, Pete
• Primary Examiner(s): Davis, Ruth A
• Application Number:
  US14/304,247
• Publication Number:
  US 20140377231A1
• Time in Patent Office: 1,712 Days
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: C12N 2506/13 from connective tissue cells, from mesenchymal cells
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage

Abstract

The present disclosure concerns methods and compositions for differentiating cells, including adipose cells, into chondrocyte-like cells via in vitro, ex vivo, and/or in vivo mechanical strain. In particular aspects, adipose cells or re-differentiated adipose cells that are chondrocyte-like cells, are delivered to a joint or are shaped into cartilage. In some embodiments, the adipose cells may be delivered to a joint, such as an intervertebral disc, following which the cells differentiate into chondrocyte-like cells to treat dysfunction of cartilage therein, including to repair degenerated discs, for example. In certain aspects, the cells prior to delivery to the individual are managed in the absence of growth factors, in vitro mechanical strain, and/or matrix molecules, for example.

Claims

1. A method of inducing de-differentiation of adipose cells to chondrocyte-like cells, comprising the steps of:

obtaining differentiated adipose cells from an individual; and

inducing de-differentiation of the differentiated adipose cells to chondrocyte-like cells by subjecting the adipose cells to mechanical strain.

2. The method of claim 1, wherein the subjecting step occurs in vitro or ex vivo.

3. The method of claim 2, wherein when the subjecting step occurs in vitro, the cells are combined with a scaffold to produce a cells/scaffold composition.

4. The method of claim 3, wherein the cells/scaffold composition comprises growth factors, matrix molecules, drugs, or a combination thereof.

5. The method of claim 3, wherein the cells/scaffold composition is delivered to an individual.

6. The method of claim 5, wherein the cells/scaffold composition is delivered to the joint of an individual.

7. The method of claim 6, wherein the joint is an intervertebral disc.

8. The method of claim 7, wherein the individual has intervertebral disc disease.

9. The method of claim 2, wherein when the subjecting step occurs ex vivo, the cells are provided conditions suitable to generate cartilage.

10. The method of claim 9, wherein the conditions comprise low oxygen, mechanical stress, or a combination thereof.

11. The method of claim 9, wherein the cartilage is configured in the form of a desired shape.

12. The method of claim 11, wherein the desired shape is at least part of an ear or of a nose.

13. The method of claim 11, further comprising the step of generating a mold of the desired shape.

14. The method of claim 11, further comprising the step of providing the cartilage to an individual that is in need of cartilage repair.

15. The method of claim 14, wherein the cartilage is provided to the individual with one or more supports.

16. The method of claim 15, wherein the support is resorbable.

17. The method of claim 15, wherein the support is comprised of a material that would be resorbed by the body of the individual during and/or after its function of cartilage formation is completed.

18. The method of claim 15, wherein the support is non-resorbable.

19. The method of claim 18, wherein the support is comprised of metal or one or more other materials that may remain in the body and promotes maintenance of shape and function of the cartilage.

20. The method of claim 14, wherein the cartilage tissue is delivered to a nose, ear, knee, shoulder, elbow or other area of the body where connective tissue is required for the individual.

21. The method of claim 14, wherein the cartilage tissue is not delivered to a joint.

22. The method of claim 14, wherein the cartilage tissue is not delivered to a vertebral disc.

23. The method of claim 11, wherein said desired shape replaces or repairs cartilage in one or more regions of the body of an individual, wherein said region requires connective tissue.

24. The method of claim 11, further comprising the step of imaging a part of the body of an individual that is in need of cartilage repair or that is at risk of being in need of cartilage repair.

25. The method of claim 11, further comprising the step of imaging a part of the body of an individual that is in need of cartilage repair and generating therefrom a mold of a desired shape of cartilage.

26. The method of claim 11, further comprising the step of imaging a part of the body of an individual wherein that part is not in need of repair and generating from that image a mold for growth of cartilage to replace or repair an area in need of repair.

27. The method of claim 1, wherein the subjecting step occurs in vivo in an individual.

28. The method of claim 27, wherein when the subjecting step occurs in vivo, the adipose cells are not subjected to growth factors, matrix molecules, mechanical strain, or a combination thereof prior to the in vivo subjecting step.

29. The method of claim 27, wherein the adipose cells are delivered to a joint of the individual.

30. The method of claim 29, wherein following delivery to the joint, there is a mixture of adipose cells and chondrocyte-like cells in the joint.

31. The method of claim 29, wherein the adipose cells are delivered between vertebral discs.

32. The method of claim 29, wherein the adipose cells are delivered between or in nucleus pulposus and fissures in the inner annulus fibrosus.

33. The method of claim 29, wherein following delivery of the adipose cells to the joint of the individual, a plurality of adipose cells die.

34. The method of claim 33, wherein death of the adipose cells results in a cellular response from endogenous joint cells of the individual.

35. The method of claim 34, wherein the cellular response comprises stimulation of growth of the endogenous joint cells of the individual.

36. The method of claim 29, wherein following delivery of the adipose cells to the joint of the individual, there is development of scar tissue in the joint.

37. The method of claim 27, wherein the individual has intervertebral disc disease.

38. The method of claim 27, wherein the cells are delivered to an intervertebral disc.

39. The method of claim 27, wherein the cells are autologous or allogeneic to the individual.

40. The method of claim 1, wherein the mechanical strain comprises low oxygen tension, intermittent hydrostatic pressure, fluid shear stress, or a combination thereof.

41. The method of claim 1, wherein the adipose cells are white adipose cells, brown adipose cells, or a mixture thereof.

42. The method of claim 1, wherein the obtained adipose cells are expanded.

43. The method of claim 42, wherein the obtained adipose cells are expanded for at least one day.

44. The method of claim 1, wherein the obtained adipose cells are passaged.

45. The method of claim 44, wherein the passaging occurs more than once.

46. A method of inducing de-differentiation of adipose cells to chondrocyte-like cells, comprising the steps of

obtaining differentiated adipose cells from an individual;

and

inducing de-differentiation of the differentiated adipose cells to chondrocyte-like cells by subjecting the adipose cells to mechanical strain, wherein the mechanical strain comprises low oxygen tension, intermittent hydrostatic pressure, fluid shear stress, or a combination thereof.

47. The method of claim 46, wherein the cells are combined with a scaffold to produce a cells/scaffold composition.

48. The method of claim 47, wherein the cells/scaffold composition is delivered to the joint of an individual.

49. The method of claim 48, wherein the joint is an intervertebral disc.

50. The method of claim 46, wherein the cells are autologous or allogeneic to the individual.