Method for inducing differentiation of pluripotent stem cells into endodermal cells

• US 10,100,284 B2
• Filed: 12/24/2014
• Issued: 10/16/2018
• Est. Priority Date: 12/25/2013
• Status: Active Grant

• Current Assignee: Toagosei Company Limited
• Original Assignee: Toagosei Company Limited
• Inventors: Tanaka, Kenichi, Yoshida, Tetsuhiko, Baileykobayashi, Nahoko
• Primary Examiner(s): Claytor, Renee
• Assistant Examiner(s): Papciak, Sharon M.
• Application Number:
  US15/107,726
• Publication Number:
  US 20160326488A1
• Time in Patent Office: 1,392 Days
• Field of Search: None
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 2319/10 containing a tag for extracellular membrane crossing, e.g. TAT or VP22
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/067 Hepatocytes

Abstract

A synthetic peptide having a stem cell differentiation-inducing activity to induce differentiation of pluripotent stem cells into endodermal cells, a stem cell differentiation inducer having this peptide as an active ingredient, and a method for inducing differentiation of pluripotent stem cells using these. The synthetic peptide provided by the present invention contains a stem cell differentiation-inducing peptide sequence, and this stem cell differentiation-inducing peptide sequence is selected from (1) an amino acid sequence constituting a signal peptide in any of amyloid precursor proteins (APP), amyloid precursor-like protein (APLP) 1 and APLP2, which are known as proteins belonging to the APP family, (2) a partial amino acid sequence constituting the signal peptide, or (3) a modified amino acid sequence formed by substitution, deletion and/or addition of 1, 2 or 3 amino acid residues in these amino acid sequences.

Claims

1. A composition for inducing differentiation of pluripotent stem cells into hepatocytes and/or liver precursor cells, comprising:

a pharmaceutically acceptable carrier; and

a synthetic peptide comprising;

a stem cell differentiation-inducing peptide sequence that induces differentiation of pluripotent stem cells into hepatocytes and/or liver precursor cells, and

a transmembrane peptide sequence at the N end of or C end of the amino acid sequence of the stem cell differentiation-inducing peptide sequence,

wherein the total number of amino acid residues constituting the synthetic peptide is 100 or less amino acid residues,

the transmembrane peptide sequence is selected from the amino acid sequences represented by any of SEQ ID NOs;

7 to 15, and

the stem cell differentiation-inducing peptide sequence is selected from the amino acid sequences of i) through iv) below;

i) Amino acid sequence of SEQ ID NO;

1 below;

2. The composition according to claim 1, wherein the synthetic peptide has the following amino acid sequence as the transmembrane peptide sequence:

3. The composition according to claim 1, wherein the total number of amino acid residues constituting the synthetic peptide is 60 or less amino acid residues.

4. The composition according to claim 1, wherein the synthetic peptide has any of the following amino acid sequences:

5. The composition according to claim 1, which increases expression, in the pluripotent stem cells, of Alpha-Fetoprotein or albumin protein, which are proteins known as markers of differentiation into liver precursor cells and hepatocytes, respectively.

6. A method for inducing differentiation of pluripotent stem cells into hepatocytes and/or liver precursor cells in vitro, the method comprising:

culturing target pluripotent stem cells; and

supplying, at least once, the composition according to claim 1 to the pluripotent stem cells in the culture.

7. The differentiation inducing method according to claim 6, wherein the pluripotent stem cells are iPS cells (induced pluripotent stem cells) derived from humans or mammals other than humans.

8. A method for producing in vitro a culture containing hepatocytes and/or liver precursor cells from a culture containing pluripotent stem cells, wherein the composition according to claim 1 is supplied at least once to the culture containing the pluripotent stem cells in a process of inducing differentiation of pluripotent stem cells into hepatocytes in the culture.