Endothelial cells derived from primate embryonic stem cells

• US 20030166273A1
• Filed: 11/01/2002
• Published: 09/04/2003
• Est. Priority Date: 11/02/2001
• Status: Active Grant

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Auerbach, Robert, Lewis, Rachel, Kaufman, Dan S.
• Granted Patent: US 7,176,023 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• 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: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells

Abstract

A method is described to induce primate embryonic stem cells to differentiate into a relatively homogenous population of endothelial cells. The ES derived endothelial cells have the general morphological and cell surface marker characteristics of endothelial cells. The ES derived endothelial cells also are capable of inducing and participating in blood vessel formation (or vascularization) when transplanted into tissue in vivo.

Claims

1. A method for directing the differentiation of primate embryonic stem cells into endothelial cells comprising the steps of

(a) culturing a culture of embryonic stem cells in a medium containing vascular endothelial cell growth factor, basic fibroblast growth factor, insulin-like growth factor and epidermal growth factor;

and

(b) sub-culturing cells which have the morphology of endothelial cells.

2. A method as claimed in claim 1 wherein the primate cells are rhesus monkey cells.

3. A method as claimed in claim 1 wherein the primate cells are human cells.

4. A method as claimed in claim 1 wherein the medium for the culture of embryonic stem cells into endothelial cells also includes mammalian serum.

5. A cell culture derived from primate embryonic stem cells, the culture comprising predominantly endothelial cells have the following characteristics:

uniform morphology of elongated to stellate shape, positive for the presence of von Willebrand factor, positive for the ability to take up acetylated LDL, and having the capability to encourage and participate in vascularization when implanted in vivo in a mammal.

6. A cell culture of endothelial cells derived from primate embryonic stem cells, the culture composed of cells over 90% of which test positive for the ability to bind the Ulex europaeus agglutinin 1 (UEA-1) lectin.