Pluripotent stem cell-derived brown adipocytes, pluripotent stem cell-derived cell aggregate, method for producing same, and cell therapy and medical therapy therefor

• US 9,492,485 B2
• Filed: 04/26/2012
• Issued: 11/15/2016
• Est. Priority Date: 04/27/2011
• Status: Active Grant

• Current Assignee: Id Pharma Co., Ltd. (I'rom Group Co., Ltd.), National Center for Global Health and Medicine
• Original Assignee: Id Pharma Co., Ltd. (I'rom Group Co., Ltd.), National Center for Global Health and Medicine
• Inventors: Saeki, Kumiko, Yuo, Akira, Nishio, Miwako, Kawasaki, Masako, Saeki, Koichi, Hasegawa, Mamoru
• Primary Examiner(s): Singh, Anoop
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US14/113,997
• Publication Number:
  US 20140140967A1
• Time in Patent Office: 1,664 Days
• US Class Current: 1/1
• 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: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/2306 Interleukin-6 (IL-6)
• CPC Class Codes: C12N 2501/26 Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue

Abstract

Provided are a method of producing brown adipocytes from pluripotent stem cells, a method of producing cell aggregates as an intermediate product thereof, pluripotent stem cell-derived cell aggregates and pluripotent stem cell-derived brown adipocytes produced by these methods, and cell therapy using the pluripotent stem cell-derived brown adipocytes. In the method of producing brown adipocytes from pluripotent stem cells, cell aggregates are produced from pluripotent stem cells by a method including the step (A), and brown adipocytes are prepared from the cell aggregates by a method including the step (B). The step (A) is a step of producing cell aggregates by non-adhesive culture of pluripotent stem cells in serum-free environment in the presence of a hematopoietic cytokine, and the step (B) is a step of producing brown adipocytes by adhesion culture of the cell aggregates in the presence of a hematopoietic cytokine.

Claims

1. A method of producing brown adipocytes, comprising the steps of:

(a) providing mouse or human pluripotent stem cells,

(b) culturing the pluripotent stem cells of step (a) in a serum-free non-adhesive culture in the presence of BMP4, VEGF, SCF, Flt3L, IL6 and IGF2 to form cell aggregates; and

(c) differentiating the cell aggregates of step (b) in an adhesive culture in the presence of BMP7, VEGF, SCF, Flt3L, IL6 and IGF3 to obtain brown adipocytes.

2. The method according to claim 1, wherein the pluripotent stem cells are ES cells or iPS cells.

3. The method according to claim 2, wherein the iPS cells are established using Sendai virus vectors encoding Oct3/4, Sox2, Klf4 and Myc.

4. The method according to claim 1, wherein the pluripotent stem cells are human pluripotent stem cells.