Somatic cell reprogramming by retroviral vectors encoding Oct3/4. Klf4, c-Myc and Sox2

• US 8,129,187 B2
• Filed: 02/18/2010
• Issued: 03/06/2012
• Est. Priority Date: 12/13/2005
• Status: Active Grant

• Current Assignee: Kyoto University
• Original Assignee: Kyoto University
• Inventors: Yamanaka, Shinya, Takahashi, Kazutoshi, Okita, Keisuke
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US12/656,907
• Publication Number:
  US 20100216236A1
• Time in Patent Office: 747 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 Klf-4
• CPC Class Codes: C12N 2501/606 c-Myc
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2740/15043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The present invention relates to a nuclear reprogramming factor having an action of reprogramming a differentiated somatic cell to derive an induced pluripotent stem (iPS) cell. The present invention also relates to the aforementioned iPS cells, methods of generating and maintaining iPS cells, and methods of using iPS cells, including screening and testing methods as well as methods of stem cell therapy. The present invention also relates to somatic cells derived by inducing differentiation of the aforementioned iPS cells.

Claims

1. A method for preparing one or more mammalian somatic cells, which comprises:

(1) introducing one or more retroviral vectors comprising the following four isolated genes;

Oct3/4, Klf4, c-Myc and Sox2 into a somatic cell obtained from a species of mammal and culturing the cell on a fibroblast feeder layer or extracellular matrix in a cell media that supports growth of ES cells of the mammalian species, wherein one or more induced pluripotent stem cells are obtained, and

(2) inducing differentiation of the induced pluripotent stem cell obtained in (1), wherein the one or more mammalian somatic cells are obtained.

3. The method of claim 1, wherein the somatic cell is a human cell.

2. A method for preparing one or more mammalian somatic cells, which comprises:

(1) introducing one or more retroviral vectors comprising the following three isolated genes;

Oct3/4, Klf4 and Sox2 into a somatic cell obtained from a species of mammal and incubating the cell in the presence of basic fibroblast growth factor on a fibroblast feeder layer or extracellular matrix in a cell media that supports growth of ES cells of the mammalian species, wherein one or more induced pluripotent stem cells are obtained, and

(2) inducing differentiation of the induced pluripotent stem cell obtained in (1), wherein the one or more mammalian somatic cells are obtained.

4. The method of claim 2, wherein the somatic cell is a human cell.