Human mesenchymal stem cells

• US 5,486,359 A
• Filed: 02/08/1994
• Issued: 01/23/1996
• Est. Priority Date: 11/16/1990
• Status: Expired due to Term

• Current Assignee: Mesoblast International Srl
• Original Assignee: Osiris Therapeutics Incorporated (Smith & Nephew PLC)
• Inventors: Caplan, Arnold I., Haynesworth, Stephen E.
• Primary Examiner(s): Knode, Marian C.
• Assistant Examiner(s): Dadio, Susan M.
• Application Number:
  US08/193,262
• Time in Patent Office: 714 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/240.25, 435/240.3, 435/240.27, 424/93 U, 424/93.7
• US Class Current: 424/93.7
• CPC Class Codes: A61F 2/28 Bones joints A61F2/30
• CPC Class Codes: A61F 2/30756 Cartilage endoprostheses A61F2/3603 takes precedence
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• 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: A61L 2430/10 for reconstruction of tendons or ligaments
• 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 27/3843 Connective tissue
• CPC Class Codes: A61L 31/005 Ingredients of undetermined constitution or reaction products thereof
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2533/18 Calcium salts, e.g. apatite, Mineral components from bones, teeth, shells
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Isolated human mesenchymal stem cells which can differentiate into more than one tissue type (e.g. bone, cartilage, muscle or marrow stroma), a method for isolating, purifying, and culturally expanding human mesenchymal stem cells (i.e. "mesenchymal stem cells" or "hMSCs"), and characterization of and uses, particularly research reagent, diagnostic and therapeutic uses for such cells. The stem cells can be culture expanded without differentiating. Monoclonal antibodies specific for human mesenchymal stem cells and the monoclonal hybridoma cell lines (ATCC nos. 10743-10745) that synthesize and secrete these monospecific antibodies, and uses of the monoclonal antibodies for diagnostic and/or therapeutic purposes.

Claims

1. An isolated, homogeneous population of human mesenchymal stem cells which can differentiate into cells of more than one connective tissue type.

2. The mesenchymal stem cells of claim 1 wherein an antibody produced from hybridoma cell line SH2, ATCC accession number HB 10743, binds to said mesenchymal stem cells.

3. The mesenchymal stem cells of claim 2 wherein said mesenchymal stem cells are obtained from bone marrow.

4. The mesenchymal stem cells of claim 3 wherein said mesenchymal stem cells adhere to a plastic surface when cultured in BGJ_b medium containing 10% fetal serum.

5. The mesenchymal stem cells of claim 3 wherein said mesenchymal stem cells retain the potential to differentiate into more than one connective tissue type after population expansion in culture.

6. The mesenchymal stem cells of claim 1 wherein an antibody produced from hybridoma cell line SH3, ATCC accession number HB 10744, binds to said mesenchymal stem cells.

7. The mesenchymal stem cells of claim 1 wherein an antibody produced from hybridoma cell line SH4, ATCC accession number HB 10745, binds to said mesenchymal stem cells.

8. The mesenchymal stem cells of claim 1 wherein said mesenchymal stem cells are obtained from bone marrow.

9. The mesenchymal stem cells of claim 8 wherein said mesenchymal stem cells adhere to a plastic surface when cultured in BGJ_b medium containing 10% fetal serum.

10. The mesenchymal stem cells of claim 1 wherein said mesenchymal stem cells are non-embryonic cells.

11. The mesenchymal stem cells of claim 1 wherein said mesenchymal stem cells adhere to a plastic surface when cultured in BGJ_b medium containing 10% fetal serum.

12. The mesenchymal stem cells of claim 1 wherein said mesenchymal stem cells retain the potential to differentiate into more than one connective tissue type after population expansion in culture.

13. The mesenchymal stem cells of claim 1 wherein said mesenchymal stem cells are obtained from periosteum.

14. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is bone.

15. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is cartilage.

16. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is adipose.

17. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is tendon.

18. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is ligament.

19. The mesenchymal stem cells of claim 1 wherein one of said connective tissue types is dermis.

20. A composition comprising the mesenchymal stem cells of claim 1 and a culture medium, wherein said culture medium expands said mesenchymal stem cells.

21. The composition of claim 20 wherein said medium is a complete medium.

22. The composition of claim 21 wherein said medium comprises BGJ_b medium.

23. A composition comprising the mesenchymal stem cells of claim 2 and a culture medium, wherein said culture medium expands said mesenchymal stem cells.

24. The composition of claim 23 wherein said medium is a complete medium.

25. The composition of claim 24 wherein said medium comprises BGJ_b medium.

26. A composition comprising the mesenchymal stem cells of claim 8 and a culture medium, wherein said culture medium expands said mesenchymal stem cells.

27. The composition of claim 26 wherein said medium is a complete medium.

28. The composition of claim 27 wherein said medium comprises BGJ_b medium.

29. A composition comprising the mesenchymal stem cells of claim 5 and a culture medium, wherein said culture medium expands said mesenchymal stem cells.

30. The composition of claim 29 wherein said medium is a complete medium.

31. The composition of claim 30 wherein said medium comprises BGJ_b medium.

32. A therapeutic composition comprising the mesenchymal stem cells of claim 1 and a pharmaceutically acceptable carrier, wherein said mesenchymal stem cells are present in an amount effective to produce connective tissue cells.

33. A therapeutic composition comprising the mesenchymal stem cells of claim 2 and a pharmaceutically acceptable carrier, wherein said mesenchymal stem cells are present in an amount effective to produce connective tissue cells.

34. A therapeutic composition comprising the mesenchymal stem cells of claim 8 and a pharmaceutically acceptable carrier, wherein said mesenchymal stem cells are present in an amount effective to produce connective tissue cells.

35. A therapeutic composition comprising the mesenchymal stem cells of claim 9 and a pharmaceutically acceptable carrier, wherein said mesenchymal stem cells are present in an amount effective to produce connective tissue cells.

36. A therapeutic composition comprising the mesenchymal stem cells of claim 11 and a pharmaceutically acceptable carrier, wherein said mesenchymal stem cells are present in an amount effective to produce connective tissue cells.

37. The therapeutic composition of claim 32 wherein said connective tissue is bone.

38. The therapeutic composition of claim 32 wherein said connective tissue is cartilage.