Method for producing neural stem cells using synthetic peptide

• US 10,421,944 B2
• Filed: 09/29/2016
• Issued: 09/24/2019
• Est. Priority Date: 09/29/2015
• Status: Active Grant

• Current Assignee: KEIO University, Toagosei Company Limited
• Original Assignee: Toagosei Company Limited
• Inventors: Baileykobayashi, Nahoko, Tanaka, Kenichi, Yoshida, Tetsuhiko, Kudo, Jun
• Primary Examiner(s): Ballard, Kimberly
• Assistant Examiner(s): MacFarlane, Stacey N
• Application Number:
  US15/279,890
• Publication Number:
  US 20170088817A1
• Time in Patent Office: 1,090 Days
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2501/90 Polysaccharides
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

The composition for producing neural stem cells provided by the present invention contains an artificially synthesized synthetic peptide having a neural stem cell-inducing peptide sequence that induces neural stem cells from fibroblasts, and one or two or more pharmaceutically acceptable carriers. The neural stem cell-inducing peptide sequence is any of (i) an amino acid sequence constituting a signal peptide of any protein belonging to the amyloid precursor protein (APP) family, (ii) a partial amino acid sequence of the amino acid sequence of (i), and (iii) a modified amino acid sequence of the amino acid sequence of (i) or (ii).

Claims

1. A method for producing neural stem cells from fibroblasts, comprising:

preparing a cell culture of the fibroblasts,

wherein the fibroblasts are cultured independently without co-cultivation with other cells; and

supplying an artificially produced synthetic peptide to the cell culture,

wherein the synthetic peptide comprises;

a) the amino acid sequence of LLLLLLVGLTAPA (SEQ ID NO;

16); and

b) a transmembrane peptide sequence of KKRTLRKNDRKKR (SEQ ID NO;

7);

and wherein the total number of amino acid residues constituting the synthetic peptide is 50 or fewer.

2. The production method according to claim 1, wherein the transmembrane peptide sequence is disposed adjacent to the N-end or C-end of the amino acid sequence of LLLLLLVGLTAPA (SEQ ID NO:

16).

3. The production method according to claim 1, wherein a linker sequence consisting of one, two or three glycine residues is disposed between the amino acid sequence of LLLLLLVGLTAPA (SEQ ID NO:

16) and the transmembrane peptide sequence.

4. The production method according to claim 1, wherein the synthetic peptide comprises the following amino acid sequence:

LLLLLLVGLTAPAGKKRTLRKNDRKKR (SEQ ID NO;

26).

5. The production method according to claim 1, wherein the fibroblasts are fibroblasts derived from human skin.

6. A method for producing neural stem cells from fibroblasts, comprising:

preparing a cell culture containing the fibroblasts; and

supplying an artificially produced synthetic peptide to the cell culture,

wherein the synthetic peptide comprises;

a) the amino acid sequence of LLLLLLVGLTAPA (SEQ ID NO;

16);

 

and

b) a transmembrane peptide sequence of KKRTLRKNDRKKR (SEQ ID NO;

7);

wherein the total number of amino acid residues constituting the synthetic peptide is 50 or fewer, and

wherein the fibroblasts are fibroblasts derived from human skin.