Method for Amplification of Endothelial Progenitor Cell in Vitro

• US 20080166327A1
• Filed: 02/22/2006
• Published: 07/10/2008
• Est. Priority Date: 02/23/2005
• Status: Active Grant

• Current Assignee: Stemmed Inc., Tokai University Educational System
• Original Assignee: Foundation for Biomedical Research and Innovation, Tokai University Educational System
• Inventors: Asahara, Takayuki, Masuda, Haruchika
• Granted Patent: US 8,492,148 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/145 Thrombopoietin [TPO]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/26 Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 5/0692 Stem cells; Progenitor cells; Precursor cells

Abstract

The present invention provides a method for expanding an endothelial progenitor cell in vitro. More particularly, the present invention provides a method for culturing a hemangioblast comprising incubating a hemangioblast in a serum-free culture medium containing one or more factors selected from the group consisting of stem cell growth factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin, and a vascular endothelial cell produced by the method; and a serum-free culture medium containing one or more factors selected from the group consisting of stem cell growth factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin, and a kit for the preparation of the serum-free culture medium and the like.

Claims

1. A method for expanding a hemangioblast, which comprises incubating the hemangioblast in a serum-free culture medium containing a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin to proliferate an endothelial progenitor cell from the hemangioblast.

3. The method of claim 1, wherein the hemangioblast is derived from bone marrow, cord blood or peripheral blood.

4. The method of claim 1, wherein the hemangioblast is a mononuclear cell.

5. The method of claim 1, wherein the hemangioblast is CD34 positive and/or CD133 positive.

6. The method of claim 1, wherein the hemangioblast and various factors to be added to the serum-free culture medium are derived from animals of the same species.

7. The method of claim 1, wherein the hemangioblast is derived from human.

8. The method of claim 1, wherein the serum-free culture medium further comprises a vascular endothelial growth factor and/or a transforming growth factor β

inhibitor.

9. An endothelial progenitor cell obtainable by the method of claim 1.

10. A composition comprising an endothelial progenitor cell obtainable by the method of claim 1, and substantially free of a biogenic substance derived from an animal of a different species from the animal, from which the endothelial progenitor cell is derived.

11. The composition of claim 10, which is used for allogeneic transplantation.

12. The composition of claim 11, wherein the endothelial progenitor cell is derived from human.

13. The composition of claim 12, which is an agent for the prophylaxis or treatment of an ischemic disease.

2. (canceled)

14. A serum-free culture medium containing a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin.

15. A kit for preparing a serum-free culture medium containing a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin, which comprises a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3, thrombopoietin and a serum-free culture medium (at least one factor from a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin is not contained in the serum-free culture medium).

16. A method for culturing a hemangioblast, which comprises incubating the hemangioblast in a serum-free culture medium containing a stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 and thrombopoietin.