Adipogenic differentiation of human mesenchymal stem cells

• US 5,827,740 A
• Filed: 07/30/1996
• Issued: 10/27/1998
• 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.
• Primary Examiner(s): Saunders, David
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US08/700,753
• Time in Patent Office: 819 Days
• Field of Search: 435/377, 435/372, 435/366, 435/405, 424/572, 424/574, 514/171, 514/46, 514/261, 514/263, 514/415, 532/2
• US Class Current: 435/372
• 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 and a compound which stimulates cAMP production or inhibits cAMP degradation (such as a phosphodiesterase inhibitor). The process can further include isolating the adipocytes from remaining hMSCs.

Claims

1. A composition which comprises mesenchymal stem cells which have been isolated from a human and which have potential to differentiate into cells of more than one connective tissue type and a substance in an amount effective to induce said stem cells to differentiate into the adipogenic lineage.

2. The composition of claim 1 wherein the substance which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage comprises a glucocorticoid and a compound which elevates intracellular levels of cAMP.

3. The composition of claim 2 wherein the glucocorticoid is selected from the group consisting of dexamethasone, hydrocortisone, and cortisone.

4. The composition of claim 2 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 2 wherein the substance which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage further comprises insulin.

8. The composition of claim 1 wherein the substance which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage comprises a glucocorticoid and a compound which inhibits degradation of cAMP.

9. The composition of claim 8 wherein the substance which inhibits degradation of cAMP is a phosphodiesterase inhibitor selected from the group consisting of methyl isobutylxanthine, theophylline, caffeine, and indomethacin.

10. The composition of claim 8 wherein the substance which induces cells from the mesenchymal stem cell population to differentiate into the adipogenic lineage further comprises insulin.

11. The process of claim 10 which further comprises isolating the adipocytes from remaining hMSCs.

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

13. The process of claim 12 wherein (i) and (ii) are in admixture with each other.

6. A process for inducing human mesenchymal stem cells to differentiate into adipocytes, comprising:

contacting human mesenchymal stem cells with (i) a glucocorticoid and (ii) a compound which elevates intracellular levels of cAMP, said glucocorticoid and said compound which elevates intracellular levels of cAMP being employed in an amount sufficient to induce human mesenchymal stem cells to differentiate into adipocytes.

7. The process of claim 6 which further comprises isolating the adipocytes from remaining hMSCs.

14. A process for inducing human mesenchymal stem cells to differentiate into adipocytes, comprising:

contacting human mesenchymal stem cells with (i) a glucocorticoid and (ii) a compound which inhibits degradation of cAMP, said glucocorticoid and said compound which inhibits degradation of cAMP being employed in an amount sufficient to induce human mesenchymal stem cells to differentiate into adipocytes.