Pluripotent stem cell growing method

• US 7,892,835 B2
• Filed: 03/23/2005
• Issued: 02/22/2011
• Est. Priority Date: 03/23/2004
• Status: Expired due to Fees

• Current Assignee: Toshihiro Akaike
• Original Assignee: Daiichi Sankyo Company Limited
• Inventors: Akaike, Toshihiro, Nagaoka, Masato, Koshimizu, Uichi, Fukuda, Keiichi
• Primary Examiner(s): Long; Scott
• Application Number:
  US10/593,831
• Publication Number:
  US 20070155013A1
• Time in Patent Office: 2,162 Days
• Field of Search: None
• 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 which exhibit a normal karyotype comprising contacting said pluripotent stem cells with a liquid medium and growing said pluripotent stem cells in said liquid medium in a dispersed state in a culturing vessel without using feeder cells, wherein said pluripotent stem cells maintain their undifferentiated state and pluripotency, said culturing vessel including immobilized or coated on a substrate solid phase surface a cadherin molecule, and wherein said pluripotent stem cells achieve cell counts at least two times greater than said pluripotent stem cells cultured on a gelatin plate after four days.

2. The method of claim 1, further comprising transferring a gene into said pluripotent stem cells, after culturing.

3. The method of claim 1, wherein said cadherin molecule 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.

4. The method of claim 3, wherein said E-cadherin is mammalian.

5. The method of claim 4, wherein said E-cadherin is human or mouse.

6. The method of claim 1, wherein said cadherin molecule is fused with an immunoglobulin Fc region and is immobilized on said substrate solid phase surface via said Fc region.

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

8. The method of claim 2, wherein said cadherin molecule is E-cadherin, or a molecule which has structure homology with said molecule, which comprises the EC1 domain and one or more domains from the EC2 domain, EC3 domain, EC4 domain and EC5 domain of E-cadherin, and which has homophilic binding ability with said pluripotent stem cells.

9. The method of claim 8, wherein said E-cadherin is mammalian.

10. The method of claim 9, wherein said E-cadherin is human or mouse.

11. The method of claim 2, wherein said cadherin molecule is fused with an immunoglobulin Fc region and is immobilized on said substrate solid phase surface via said Fc region.

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

13. The method of claim 2, wherein said cadherin molecule is a human or a mouse E-cadherin and said pluripotent stem cells are mammalian embryonic stem cells (ES cells).