Method for preparing pluripotent stem cells

• US 9,765,296 B2
• Filed: 09/03/2014
• Issued: 09/19/2017
• Est. Priority Date: 09/04/2013
• Status: Active Grant

• Current Assignee: Otsuka Pharmaceutical Factory Incorporated (Otsuka Holdings Company Limited)
• Original Assignee: Otsuka Pharmaceutical Factory Incorporated (Otsuka Holdings Company Limited)
• Inventors: Okairi, Risa, Nishimura, Masuhiro, Wada, Tamaki
• Primary Examiner(s): BERKE-SCHLESSEL, DAVID W
• Application Number:
  US14/913,707
• Publication Number:
  US 20160215258A1
• Time in Patent Office: 1,112 Days
• Field of Search: None
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 2506/1384 from adipose-derived stem cells [ADSC], from adipose stromal stem cells
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

An object of the present invention is to provide a method capable of inexpensively and conveniently preparing cells having pluripotency and a very low risk of tumorigenic transformation. The cells having pluripotency and a very low risk of tumorigenic transformation can be prepared by suspension-culturing mammalian mesenchymal stem cells such as human mesenchymal stem cells from bone marrow (hMSC-BM) and human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also referred to as “human adipose-derived stem cells [hADSC]”), 7 types of human adherent mature cells (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal lymphatic microvascular endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]), and 3 types of human adherent precursor cells (human dermal fibroblast cells [NHDF cells], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) to form cell masses (spheroids).

Claims

1. A method for preparing a pluripotent stem cell, comprising spheroid-culturing mammalian mesenchymal stem cells to form a spheroid of pluripotent stem cells,

wherein the spheroid-culturing is performed in a solution containing the following (A) or (B):

(A) gellan gum or a derivative thereof or a salt of these;

or

(B) dextran or a derivative thereof or a salt of these.

2. The method according to claim 1, wherein the mammalian mesenchymal stem cells are human mesenchymal stem cells from bone marrow or human adipose tissue-derived mesenchymal stem cells.

3. The method according to claim 1, wherein the pluripotent stem cell expresses Nanog, Oct3/4, or Sox2.

4. The method according to claim 2, wherein the pluripotent stem cell expresses Nanog, Oct3/4, or Sox2.