Human pluripotent stem cells induced from undifferentiated stem cells derived from a human postnatal tissue

• US 9,714,433 B2
• Filed: 03/03/2014
• Issued: 07/25/2017
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Kyoto University
• Original Assignee: Kyoto University
• Inventors: Sakurada, Kazuhiro, Ishikawa, Tetsuya, Masaki, Hideki, Takahashi, Shunichi
• Primary Examiner(s): Ton, Thaian N
• Application Number:
  US14/195,698
• Publication Number:
  US 20140206083A1
• Time in Patent Office: 1,240 Days
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 15/85 for animal cells
• 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 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

Establishment of human pluripotent stem cells having properties close to human ES cells with the genome of the patient per se that can circumvent immunological rejection of transplanted cells from cells derived from a postnatal human tissue are described. Human pluripotent stem cells can be induced by introducing three genes of Oct3/4, Sox2 and Klf 4, or three genes of Oct3/4, Sox2 and Klf 4 plus the c-Myc gene or a histone deacetylase (HDAC) inhibitor of undifferentiated stem cells present in various human postnatal tissues in which each gene of Tert, Nanog, Oct3/4 and Sox2 has not undergone epigenetic inactivation.

Claims

1. A method for generating a human pluripotent stem cell, comprising the steps of:

(i) introducing one or more retroviral vectors comprising a gene encoding Oct3/4, a gene encoding Sox2 and a gene encoding Klf4 into an isolated human post-natal cell; and

(ii) culturing the cell of step (i) in the presence of an inhibitor of Rho kinase and/or a histone deacetylase inhibitor under conditions that maintain pluripotency and self-renewal, thereby generating a human pluripotent stem cell.

2. The method of claim 1, wherein the inhibitor of Rho kinase is Y-27632 or HA-1077.