Method for amplification of endothelial progenitor cell in vitro

• US 8,492,148 B2
• Filed: 02/22/2006
• Issued: 07/23/2013
• 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
• Primary Examiner(s): DRISCOLL, LORA E BARNHART
• Application Number:
  US11/884,949
• Publication Number:
  US 20080166327A1
• Time in Patent Office: 2,708 Days
• Field of Search: None
• US Class Current: 435/377
• 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 ligand and thrombopoietin, and a kit for the preparation of the serum-free culture medium and the like.

Claims

1. A method for preparing endothelial progenitor cells, which comprises incubating CD34 positive and/or CD133 positive cells in a serum-free culture medium containing stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 ligand, thrombopoietin and vascular endothelial growth factor, whereby the number of endothelial progenitor cells contained in the CD34 positive and/or CD133 positive cells is increased.

2. The method of claim 1, wherein the CD34 positive and/or CD133 positive cells include mononuclear cells including endothelial progenitor cells.

3. The method of claim 2, wherein the CD34 positive and/or CD133 positive cells are prepared from bone marrow fluid, cord blood or peripheral blood.

4. The method of claim 1, wherein the CD34 positive and/or CD133 positive cells include endothelial progenitor cells.

5. The method of claim 4, wherein the CD34 positive and/or CD133 positive cells are prepared from bone marrow fluid, cord blood or peripheral blood.

6. The method of claim 1, wherein the CD34 positive and/or CD133 positive cells, stem cell factor, interleukin-6, FMS-like tyrosine kinase 3 ligand, thrombopoietin, and vascular endothelial growth factor are all obtained from animals of a single species.

7. The method of claim 1, wherein the CD34 positive and/or CD133 positive cells are derived from human.

8. The method of claim 1, wherein the serum-free culture medium further comprises transforming growth factor β

inhibitor.