Methods for nuclear reprogramming of cells

Patent metadata

Field	Value
Publication number	US-10745668-B2
Application number	US-201414493107-A
Country	US
Kind code	B2
Filing date	Sep 22, 2014
Priority date	Sep 20, 2013
Publication date	Aug 18, 2020
Grant date	Aug 18, 2020

Abstract

Described herein are methods for enhancing the nuclear reprogramming of somatic cells to become induced pluripotent stem cells. In particular, the methods disclosed herein involve the use of damage-associated molecular pattern molecules (DAMP). In certain embodiments the DAMPs are aluminum compositions such as aluminum hydroxide. Such DAMPs have unexpectedly and surprisingly been found to enhance the nuclear reprogramming efficiency of the reprogramming factors commonly used to induce somatic cells to become induced pluripotent stem cells. Accordingly, this disclosure describes methods of nuclear reprogramming as well as cells obtained from such methods along with therapeutic methods for using such cells for the treatment of disease amendable to treatment by stem cell therapy; as well as kits for such uses.

First claim

What is claimed is:

1. A method of nuclear reprogramming of a human somatic cell in vitro, the method comprising:
(a) contacting a population of human somatic cells with an effective dose of an aluminum hydroxide gel composition, wherein the effective dose of aluminum hydroxide is at least 40-80 micrograms/ml;
(b) transfecting said human somatic cells with a plasmid comprising a nucleic acid encoding nuclear reprogramming factor Oct3/4;
(c) transferring the transfected cells of step (b) into a vitronectin coated plates placed in incubator for at least 2 days:
(d) suspending the cells of step (c) into a pluripotent culture medium;
(e) culturing the cells of step (d) into vitronectin coated plates for at least 2 days;
(f) adding pluripotent culture medium to the plates every other day for a period of time sufficient until colonies appear; thereby reprogramming said human somatic cells to human induced pluripotent stem (iPSC) cells, wherein the nuclear reprogramming efficiency is greater than if the method was carried out without the aluminum hydroxide gel composition.

2. The method of claim 1 , wherein the aluminum hydroxide gel composition and reprogramming factors are provided sequentially or simultaneously.

3. The method of claim 1 , wherein the human somatic cell is a peripheral blood mononuclear cell (PBMC), a cord blood mononuclear cell, or a fibroblast.

Assignees

• Lonza Ag

Inventors

• Walsh Patrick
• Fellner Thomas

Classifications

• C12N2533/52

  Fibronectin; Laminin · CPC title

• A61P25/16

  Anti-Parkinson drugs · CPC title

• C12N2500/12

  Light metals, i.e. alkali, alkaline earth, Be, Al, Mg · CPC title

• C12N5/0696 Primary

  Artificially induced pluripotent stem cells, e.g. iPS · CPC title

• A61P25/28

  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 title