Lineage-directed induction of human mesenchymal stem cell differentiation

• US 5,736,396 A
• Filed: 01/24/1995
• Issued: 04/07/1998
• Est. Priority Date: 01/24/1995
• Status: Expired due to Term

• Current Assignee: Osiris Therapeutics Incorporated (Smith & Nephew PLC)
• Original Assignee: Case Western Reserve University
• Inventors: Bruder, Scott P., Caplan, Arnold I., Haynesworth, Stephen E.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Application Number:
  US08/377,461
• Time in Patent Office: 1,169 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/240.3, 435/240.31, 435/366, 435/372, 514/2, 424/93.7
• US Class Current: 435/366
• 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 31/203 Retinoic acids ; Salts thereof
• CPC Class Codes: A61K 31/557 Eicosanoids, e.g. leukotrienes or prostaglandins
• CPC Class Codes: A61K 31/573 substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
• CPC Class Codes: A61K 31/7068 having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
• 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 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 38/1841 Transforming growth factor [TGF]
• CPC Class Codes: A61K 38/1875 Bone morphogenic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: A61K 38/2006 IL-1
• CPC Class Codes: A61K 38/2013 IL-2
• CPC Class Codes: A61K 38/22 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin A61K38/33, e.g. corticotropin A61K38/35
• CPC Class Codes: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2501/06 Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/2301 Interleukin-1 (IL-1)
• CPC Class Codes: C12N 2501/39 : Steroid hormones
• CPC Class Codes: C12N 5/0654 : Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0655 : Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0658 : Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts
• CPC Class Codes: C12N 5/0663 : Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Methods for in vitro or ex vivo lineage directed induction of isolated, culture expanded human mesenchymal stem cells comprising contacting the mesenchymal stem cells with a bioactive factor effective to induce differentiation thereof into a lineage of choice as well as such compositions including isolated culture expanded human mesenchymal stem cells and bioactive factors effective to induce directed lineage induction are disclosed. Further disclosed is this method which also includes introducing such culturally expanded lineage-induced mesenchymal stem cells into a host from which they have originated for purposes of mesenchymal tissue regeneration or repair.

Claims

1. A method of inducing ex vivo lineage-directed differentiation of isolated human mesenchymal stem cells which comprises contacting the mesenchymal stem cells with a bioactive factor so as to thereby induce ex vivo differentiation thereof into a single particular mesenchymal lineage.

2. The method of claim 1 wherein the bioactive factor induces differentiation of such cells into a mesenchymal lineage selected from the group consisting of osteogenic, chondrogenic, tendonogenic, ligamentogenic, myogenic, marrow stromagenic, adipogenic and dermogenic.

3. The method of claim 1 wherein the cells are contacted with the bioactive factor and differentiated ex vivo in a rigid porous vessel.

4. The method of claim 3 wherein the rigid porous vessel is a ceramic cube.

5. The method of claim 1 wherein the cells are contacted with the bioactive factor and differentiated ex vivo in a culture vessel.

6. The method of claim 5 wherein the culture vessel is formed of a material selected from the group consisting of glass and plastic.

7. The method of claim 1 wherein the cells are contacted with the bioactive factor in a pharmaceutically acceptable liquid.

8. The method of claim 7 wherein the liquid is suitable for intranmuscular, intravenous or intraarticular injection.

9. The method of claim 1 which comprises inducing ex vivo osteogenic lineage differentiation and the bioactive factor is an osteoinductive factor.

10. The method of claim 9 wherein the osteoinductive factor is a bone morphogenic protein.

11. The method of claim 10 wherein the bone morphogenic protein is selected from the group consisting of BMP-2 and BMP-3.

12. The method of claim 9 wherein the osteoinductive factor is a fibroblast growth factor.

13. The method of claim 12 wherein the fibroblast growth factor is basic fibroblast growth factor.

14. The method of claim 9 wherein the osteoinductive factor is a glucocorticoid.

15. The method of claim 14 wherein the glucocorticoid is dexamethasone.

16. The method of claim 9 wherein the osteoinductive factor is a prostaglandin.

17. The method of claim 16 wherein the prostaglandin is prostaglandin El.

18. The method of claim 9 which further comprises contacting the isolated human mesenchymal stem cells with an adjunct factor.

19. The method of claim 18 wherein the adjunct factor is selected from the group consisting of ascorbic acid and its analogs and a glycerophosphate.

20. The method of claim 1 which comprises inducing ex vivo chondrogenic lineage differentiation and the bioactive factor is a chondroinductive factor.

21. The method of claim 20 wherein the chondroinductive factor is a member of the transforming growth factor-β

superfamily.

22. The method of claim 21 wherein the transforming growth factor-β

superfamily member is TGF-β

1.

23. The method of claim 21 wherein the transforming growth factor-β

superfamily member is inhibin A.

24. The method of claim 21 wherein the transforming growth factor-β

superfamily member is chondrogenic stimulating activity factor.

25. The method of claim 21 wherein the transforming growth factor-β

superfamily member is a bone morphogenic protein.

26. The method of claim 25 wherein the bone morphogenic protein is BMP-4.

27. The method of claim 20 wherein the chondroinductive factor is a component of the collagenous extracellular matrix.

28. The method of claim 27 wherein the collagenous extracellular matrix component is collagen I.

29. The method of claim 28 wherein the collagen I is in the form of a gel.

30. The method of claim 20 wherein the chondroinductive factor is a vitamin A analog.

31. The method of claim 30 wherein the vitamin A analog is retinoic acid.

32. The method of claim 1 which comprises inducing ex vivo stromagenic lineage differentiation and the bioactive factor is a stromainductive factor.

33. The method of claim 32 wherein the stromainductive factor is an interleukin.

34. The method of claim 33 wherein the interleukin is selected from the group consisting of interleukin-1α

and interleukin-2.

35. The method of claim 1 which comprises inducing ex vivo myogenic lineage differentiation and the bioactive factor is a myoinductive factor.

36. The method of claim 35 wherein the myoinductive factor is a cytidine analog.

37. The method of claim 36 wherein the cytidine analog is selected from the group consisting of 5-azacytidine and 5-aza-2'"'"'-deoxycytidine.

38. A method of treating an individual in need of mesenchymal cells of a particular mesenchymal lineage which comprises administering to an individual in need thereof a composition comprising isolated, human mesenchymal stem cells which have been induced to differentiate ex vivo by contact with a bioactive factor so as to thereby induce ex vivo differentiation of such cells into a single particular mesenchymal lineage.