Pluripotent stem cell growing method

• US 20070155013A1
• Filed: 03/23/2005
• Published: 07/05/2007
• Est. Priority Date: 03/23/2004
• Status: Active Grant

• Current Assignee: Toshihiro Akaike
• Original Assignee: Asubio Pharma Co., Ltd. (Daiichi Sankyo Company Limited), Keiichi Fukuda, Toshihiro Akaike
• Inventors: Akaike, Toshihiro, Nagaoka, Masato, Koshimizu, Uichi, Fukuda, Keiichi
• Granted Patent: US 7,892,835 B2
• Time in Patent Office: Days
• US Class Current: 435/455
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

A novel growing method is provided for pluripotent stem cells such as ES cells. The method of the invention is a pluripotent stem cell growing method and gene transfer method in which pluripotent stem cells are cultured under conditions that maintain their undifferentiated state and pluripotency, the method being characterized by using a liquid medium and a culturing vessel having immobilized or coated on a substrate solid phase surface a molecule which is adhesive to the pluripotent stem cells in a fixed concentration, to grow the pluripotent stem cells in a dispersed state while maintaining their undifferentiated state and pluripotency, without using feeder cells, or to transfer and express a gene therein.

Claims

1. A method for growing pluripotent stem cells, comprising growing said pluripotent stem cells in a dispersed state while maintaining their undifferentiated state and pluripotency, in a liquid medium and a culturing vessel including immobilized or coated on a substrate solid phase surface a molecule which is adhesive to said pluripotent stem cells, without using feeder cells.

3. The method of claim 1 or 2, wherein the molecule which is adhesive to said pluripotent stem cells is either a molecule that is expressed by said pluripotent stem cells or a molecule that is structurally homologous with said molecule and has homophilic binding ability with said pluripotent stem cells.

4. The method of claim 3, wherein the molecule which is adhesive to said pluripotent stem cells is a molecule belonging to the cadherin family.

5. The method of claim 4, wherein said molecule belonging to the cadherin family is E-cadherin, or a molecule which has structural homology with said molecule, which comprises the EC1 domain and one or more domains from among the EC2 domain, EC3 domain, EC4 domain and EC5 domain of E-cadherin, and which has homophilic binding ability with said pluripotent stem cells.

6. The method of claim 5, wherein said E-cadherin is obtained from a mammal.

7. The method of claim 6, wherein said E-cadherin is obtained from a human or mouse.

8. The method of claim 1 or 2, wherein the molecule which is adhesive to said pluripotent stem cells is fused with an immunoglobulin Fc region and is immobilized on said substrate solid phase surface via said Fc region.

9. The method of claim 1 or 2, wherein said pluripotent stem cells are mammalian embryonic stem cells (ES cells) or embryonic germ cells (EG cells).

10. Pluripotent stem cells produced by the method of claim 1 or 2.

2. A method for gene transfer in pluripotent stem cells, comprising transferring a gene into said pluripotent stem cells, and growing said pluripotent stem cells in a liquid medium and a culturing vessel including immobilized or coated on a substrate solid-phase surface a molecule which is adhesive to said pluripotent stem cells, without using feeder cells.