Chemically defined medium for human mesenchymal stem cells

• US 5,908,782 A
• Filed: 06/05/1995
• Issued: 06/01/1999
• Est. Priority Date: 06/05/1995
• Status: Expired due to Term

• Current Assignee: Mesoblast International Srl
• Original Assignee: Osiris Therapeutics Incorporated (Smith & Nephew PLC)
• Inventors: Marshak, Daniel R., Holecek, James J.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Assistant Examiner(s): TATE, CHRISTOPHER ROBIN
• Application Number:
  US08/464,599
• Time in Patent Office: 1,457 Days
• Field of Search: 424/93.7, 424/93.1, 435/69.4, 435/70.3, 435/240.1, 435/240.2, 435/240.3, 435/240.21, 435/366, 435/371-372, 435/383, 435/388, 435/384, 435/389, 435/404, 435/405, 435/407, 530/363
• US Class Current: 435/366
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/825 Serotonine (5-HT); Melatonine
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

A composition and method for maintaining the viability of human mesenchymal precursor cells in a serum-free environment which composition includes (1) a minimum essential medium; (2) serum albumin; (3) an iron source; (4) insulin or an insulin-like growth factor; and (5) at least one amino acid selected from the group consisting of glutamine, arginine and and cysteine, and is free of serum. Also, a composition and method for culture expanding human mesenchymal precursor cells in a serum-free environment. This composition further includes a mitogen, paricularly a serotonergic agonist. The cells are preferably isolated human mesenchymal stem cells.

Claims

1. A method for culturally expanding human mesenchymal stem cells comprising culturing an isolated homogenous population of viable human mesenchymal stem cells for a suitable time in a serum free medium which comprises (1) a minimum essential medium;

(2) serum albumin;

(3) an iron source;

(4) insulin or an insulin-like growth factor;

(5) glutamine; and

(6) a mitogen selected from the group consisting of platelet derived growth factor and serotonin, which are present in amounts effective to culturally expand the human mesenchymal stem cells.

2. The method of claim 1 wherein the iron source is transferrin.

3. The method of claim 1 wherein the insulin-like growth factor is IGF-1 or IGF-2.

4. The method of claim 1 wherein the serum free medium further includes a lipid.

5. The method of claim 1 wherein the serum free medium further includes an antioxidant.

6. The method of claim 5 wherein the antioxidant is ascorbic acid or an analog or derivative thereof.

7. A composition which comprises an isolated homogenous population of viable human mesenchymal stem cells in a serum free medium comprising (1) a minimum essential medium;

(2) serum albumin;

(3) an iron source;

(4) insulin or an insulin-like growth factor;

(5) glutamine; and

(6) a mitogen selected from the group consisting of platelet derived growth factor and serotonin which are present in amounts effective to culturally expand the human mesenchymal stem cells in the absence of serum.

8. The composition of claim 7 wherein the iron source is transferrin.

9. The composition of claim 7 wherein the insulin-like growth factor is IGF-1 or IGF-2.

10. The composition of claim 7 wherein the composition further includes a lipid.

11. The composition of claim 7 which further includes an antioxidant.

12. The composition of claim 11 wherein the antioxidant is ascorbic acid or an analog or derivative thereof.