Production method for neuron-like cells

Patent metadata

Field	Value
Publication number	US-12415986-B2
Application number	US-201916972834-A
Country	US
Kind code	B2
Filing date	Jul 5, 2019
Priority date	Jul 10, 2018
Publication date	Sep 16, 2025
Grant date	Sep 16, 2025

Abstract

It is a main object of the present invention to provide a process for producing a neuron-like cell from a somatic cell, a neuron-like cell obtained thereby, and a composition comprising a combination of chemical substances that can be used for said process without performing artificial gene transfer.
The invention can include, for example, a process for producing neuron-like cells, characterized by comprising a step of culturing somatic cells in the presence of two kinds of inhibitors, i.e., a TGF-β inhibitor and a BMP inhibitor, as well as any three or more selected from the group consisting of four kinds, i.e., a cAMP inducer, a GSK3 inhibitor, an Erk-inhibitor, and a p53-inhibitor, wherein the TGF-β inhibitor is a selective ALK5 inhibitor, and a neuron-like cell obtained by said process.
The neuron-like cells obtained according to the present invention are useful in regenerative medicine and the like.

First claim

The invention claimed is:

1. A process for producing a neuron-like cell by direct differentiation induction from a skin fibroblast, comprising
a step of culturing the skin fibroblast in the presence of a TGF-β inhibitor, a BMP inhibitor, a cAMP inducer, and a GSK3 inhibitor and in the absence of a p53 inhibitor and a growth factor, wherein
the TGF-β inhibitor comprises RepSox (CAS No: 446859-33-2),
the BMP inhibitor comprises LDN193189 and dorsomorphin, and
the process directly induces differentiation from the fibroblast to the neuron-like cell without generating a stem cell.

2. The process for producing a neuron-like cell according to claim 1 , wherein the BMP inhibitor comprises an ALK2/3 inhibitor.

3. The process for producing a neuron-like cell according to claim 1 , wherein the GSK3 inhibitor comprises CHIR99021 (CAS No: 252917-06-9).

4. The process for producing a neuron-like cell according to claim 1 , wherein the skin fibroblast is cultured in the absence of a cytokine.

5. The process for producing a neuron-like cell according to claim 1 , wherein the CAMP inducer comprises forskolin.

6. The process for producing a neuron-like cell according to claim 1 , wherein the TGF-β inhibitor comprises an inhibitor which exclusively inhibits ALK5.

7. The process for producing a neuron-like cell according to claim 1 , wherein the skin fibroblast is cultured in the absence of a histone deacetylase inhibitor.

8. The process for producing a neuron-like cell according to claim 7 , wherein the histone deacetylase inhibitor is valproic acid.

Assignees

• Kataoka Corp
• Kyoto Prefectural Public Univ Corp

Inventors

• Dai Ping
• Harada Yoshinori
• Matsumoto Junichi

Classifications

• C12N2506/1307

  from adult fibroblasts · CPC title

• C12N2501/40

  Regulators of development · CPC title

• C12N2501/01

  Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin · CPC title

• C12N2501/155

  Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor · CPC title

• C12N2501/15

  Transforming growth factor beta (TGF-β) · CPC title