Culturing of mesenchymal stem cells with FGF4 and nicotinamide

• US 10,047,345 B2
• Filed: 02/13/2013
• Issued: 08/14/2018
• Est. Priority Date: 02/13/2012
• Status: Expired due to Fees

• Current Assignee: Gamida Cell, Ltd.
• Original Assignee: Gamida Cell, Ltd.
• Inventors: Peled, Tony, Steinhardt, Yair
• Primary Examiner(s): Lankford, Blaine
• Application Number:
  US14/378,235
• Publication Number:
  US 20150004146A1
• Time in Patent Office: 2,008 Days
• Field of Search: None
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• 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: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2502/1358 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 2502/1382 Adipose-derived stem cells [ADSC], adipose stromal stem cells
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

Methods of culturing mesenchymal stem cells are provided. The methods comprise culturing MSCs in a medium comprising nicotinamide and fibroblast growth factor 4 (FGF4). Populations of mesenchymal stem cells generated using the methods described herein and uses thereof are also provided.

Claims

1. A method of expanding undifferentiated mesenchymal stem cells (MSCs) comprising culturing a population of MSCs comprising undifferentiated MSCs in a medium comprising nicotinamide and fibroblast growth factor 4 (FGF4) wherein said medium is devoid of basic fibroblast growth factor, (bFGF), platelet derived growth factor (PDGF) and erythrocyte growth factor (EGF), thereby generating an expanded population of cultured undifferentiated MSCs.

2. The method of claim 1, wherein said medium comprises DMEM.

3. The method of claim 1, wherein said medium comprises serum or platelet lysate.

4. The method of claim 1, wherein the mesenchymal stem cells are derived from a tissue selected from the group consisting of bone marrow, adipose tissue, placenta and umbilical cord blood.

5. The method of claim 1, wherein said nicotinamide is selected from the group consisting of nicotinamide, a nicotinamide analog, a nicotinamide metabolite, a nicotinamide analog metabolite and derivatives thereof.

6. The method of claim 1, wherein said culturing is effected on a plastic surface.

7. The method of claim 1, wherein said population of MSCs is comprised in a heterogeneous population of cells.

8. The method of claim 1, wherein a calcium concentration of said medium is greater than 1.8 mM.

9. The method of claim 1, wherein a concentration of said nicotinamide is 1-20 mM.

10. The method of claim 1, wherein said medium is devoid of platelet derived growth factor (PDGF).

11. The method of claim 1, wherein said undifferentiated MSC are a seeded population of mesenchymal stem cells cultured in a medium devoid of nicotinamide prior to said culturing in the medium comprising nicotinamide and FGF4.

12. The method of claim 11, wherein said culturing in said medium comprising nicotinamide and FGF-4 is effected for at least one week.

13. The method of claim 11, wherein said culturing in said medium comprising nicotinamide and FGF-4 or said medium devoid of nicotinamide is effected in a medium comprising calcium, wherein a concentration of said calcium is greater than 1.8 mM.