Adipogenic differentiation of human mesenchymal stem cells

• US 6,322,784 B1
• Filed: 10/26/1998
• Issued: 11/27/2001
• Est. Priority Date: 07/30/1996
• Status: Expired due to Term

• Current Assignee: Mesoblast International Srl
• Original Assignee: Osiris Therapeutics Incorporated (Smith & Nephew PLC)
• Inventors: Pittenger, Mark F., Beck, Stephen C.
• Primary Examiner(s): Chan, Christine Y.
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US09/246,003
• Time in Patent Office: 1,128 Days
• Field of Search: 424/93.7, 514/3, 514/46, 514/169, 514/171, 435/325, 435/366, 435/372, 435/377, 435/405, 435/395, 530/303, 552/502, 562/503
• US Class Current: 424/93.7
• 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: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

A composition which comprises human mesenchymal stem cells which have the potential to differentiate into cells of more than one connective tissue type and a composition which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage, and a process for inducing such differentiation. The composition for inducing such differentiation comprises a glucocorticoid, a compound which stimulates cAMP production or inhibits cAMP degradation (such as a phosphodiesterase inhibitor), and/or a compound which upregulates peroxisome proliferator activated receptor γ (PPAR γ) expression and/or increases its binding affinity to its DNA binding site. The process can further include isolating the adipocytes from remaining hMSCs.

Claims

1. A composition which comprises human mesenchymal stem cells (hMSCs) and a substance which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage wherein the substance comprises (i) a glucocorticoid;

(ii) a member selected from the group consisting of (a) a compound which elevates intracellular levels of cAMP and (b) a compound which inhibits degradation of cAMP; and

(iii) a compound which upregulates peroxisome proliferator activated receptor γ

(PPAR γ

) expression and/or increases its binding affinity to its DNA binding site.

2. The composition of claim 1, further comprising insulin.

3. The composition of claim 1, wherein the glucocorticoid is selected from the group consisting of dexamethasone, hydrocortisone, and cortisone.

4. The composition of claim 1, wherein the substance which elevates intracellular cAMP is selected from the group consisting of dibutyryl-cAMP, 8-CPT-cAMP (8-(4)-chlorophenylthio)-adenosine 3′

, 5′

cyclic monophosphate;

8-bromo-cAMP;

dioctanoyl-cAMP; and

Forskolin.

5. The composition of claim 1, wherein the substance which inhibits degradation of cAMP is a phosphodiesterase inhibitor selected from the group consisting of methyl isobutylxanthine, theophylline, caffeine, and indomethacin.

6. The composition of claim 1 where in the substance which upregulates peroxisome proliferator activated receptor γ

(PPARγ

) expression and/or increases its binding affinity to its DNA binding site is a prostaglandin.

7. The composition of claim 6 wherein the substance which upregulates peroxisome proliferator activated receptor γ

(PPAR γ

) expression and/or increases its binding affinity to its DNA binding site is 15-deoxy Δ

^12,14-prostaglandin J₂ (15d-PGJ₂).

8. A process for inducing a human mesenchymal stem cells (hMSCs) to differentiate into adipocytes, comprising:

contacting human mesenchymal stem cells with (i) a glucocorticoid;

(ii) a member selected from the group consisting of (a) a compound which elevates intracellular levels of cAMP and (b) a compound which inhibits degradation of cAMP; and

(iii) a compound which upregulates peroxisome proliferator activated receptor 65 (PPAR γ

) expression and/or increases its binding affinity to its DNA binding site in an amount sufficient to induce human mesenchymal stem cells to differentiate into adipocytes.

9. The process of claim 8, further comprising contacting the cells with insulin.

10. The process of claim 8, wherein (ii) is a compound which inhibits a compound which degrades cAMP.

11. The process of claim 10, wherein (ii) is a phosphodiesterase inhibitor.

12. The process of claim 8, which further comprises isolating the adipocytes from remaining hMSCs.