GROWTH FACTOR COCKTAIL TO ENHANCE OSTEOGENIC DIFFERENTIATION OF MESENCHYMAL CELLS

• US 20150307846A1
• Filed: 06/19/2013
• Published: 10/29/2015
• Est. Priority Date: 06/19/2012
• Status: Abandoned Application

• Current Assignee: Katholieke Universiteit Leuven, University of Pennsylvania
• Original Assignee: Katholieke Universiteit Leuven, University of Pennsylvania
• Inventors: Luyten, Frank, Chen, Christopher, Eyckmans, Jeroen, Schrooten, Jan, Roberts, Scott
• Application Number:
  US14/408,416
• Publication Number:
  US 20150307846A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/2306 Interleukin-6 (IL-6)
• CPC Class Codes: C12N 2501/25 Tumour necrosing factors [TNF]
• CPC Class Codes: C12N 2506/13 from connective tissue cells, from mesenchymal cells
• CPC Class Codes: C12N 2506/1392 from mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/0654 Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth

Abstract

The present invention relates to methods and compositions for osteogenic differentiation of human periosteum derived cells, in particular using a growth medium containing a specific combination of growth factors and formulations thereof. The invention also relates to the differentiated cells and cell populations, as well as further products comprising such cells and uses thereof in bone therapy.

Claims

1. A method for inducing cells to proliferate and differentiate into cells with a osteogenic phenotype, the method comprising culturing cells in a medium comprising about 2 ng/ml to about 200 ng/ml EGF, about 1 ng/ml to about 100 ng/ml IL6, and about 1 ng/ml to about 100 ng/ml TGFβ

1.

2. The method of claim 1 , wherein the medium comprises about 20 ng/ml EGF, about 10 ng/ml IL6, and about 10 ng/ml TGFβ

1.

3. The method of claim 1, wherein the medium contains a calcium ion concentration ranging from about 0.3 mM to about 12 mM.

5. The method of claim 1, wherein the medium contains a serum concentration ranging from 0% to about 20%.

7. The method of claim 1, wherein the medium contains from about 10⁻

4 M to about 10<sup>−

7</sup> M ascorbic acid.

9. The method of claim 1, wherein the medium contains a phosphate ion concentration ranging from about 0.2 mM to about 8 mM.

11. The method of claim 1, wherein the cells are cultured for at least four days.

13. The method of claim 1, wherein the cells are cultured in a medium which additionally comprises TNFα

in a first period, wherein said first period is maximum 4 days.

15. The method of claim 1, wherein the cells that are cultured with the medium comprising EGF, IL6 and TGFβ

1 are stem cells.

21. Cells produced according to the method recited in claim 1.

30. A pharmaceutical composition comprising the cells produced according to the method recited in claim 1.

31. A method of treatment comprising administering a therapeutically effective amount of the cells produced according the method recited in claim 1 to a subject with a bone disorder.

34. The method of claim 31, wherein said bone disorder is a bone fracture or a non healing bone defect.

35. The method of claim 31, wherein the subject is a human patient.

36. The method of claim 31, further comprising administering non-cellular material to said subject.

37. The method of claim 36, wherein the cells and the non-cellular material are combined in vitro to form an implantable graft.

4. (canceled)

6. (canceled)

8. (canceled)

10. (canceled)

12. (canceled)

14. (canceled)

16-20. -20. (canceled)

22. A composition, comprising cells in a culture medium comprising about 2 ng/ml to about 200 ng/ml EGF, about 1 ng/ml to about 100 ng/ml IL6 and about 1 ng/ml to about 100 ng/ml TGFβ

1, wherein the cells express a primitive mesenchymal phenotype in the culture medium.

23. The composition of claim 22, wherein the medium is comprised of about 20 ng/ml EGF, about 10 ng/ml IL6 and about 10 ng/ml TGFβ

1.

24. The composition of claim 22, wherein the medium further comprises serum in a concentration from 0% to about 20%.

26. The composition of claim 22, wherein the medium further comprises about 10⁻

4 M to about 10<sup>−

7</sup> M ascorbic acid.

25. (canceled)

27-29. -29. (canceled)

32-33. -33. (canceled)