Induced pluripotent stem cells produced with Oct3/4, Klf4 and Sox2

• US 8,278,104 B2
• Filed: 06/13/2008
• Issued: 10/02/2012
• 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:
  US12/213,035
• Publication Number:
  US 20090047263A1
• Time in Patent Office: 1,572 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 2501/60 Transcription factors
• 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 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 an induced pluripotent stem cell by nuclear reprogramming of a mammalian somatic cell, comprising:

a) introducing into the mammalian somatic cell one or more retroviral vectors comprising Oct3/4, Klf4 and Sox2 operably linked to a promoter, and

b) culturing the transduced somatic cell in the presence of a cytokine on a fibroblast feeder layer or extracellular matrix under conditions that maintain pluripotency and self renewal.

2. The method according to claim 1, wherein the mammalian somatic cell is a human cell.

3. The method according to claim 1, wherein the mammalian somatic cell is a mouse cell.

4. The method according to claim 1, wherein the cytokine is bFGF and/or SCF.