Method for inducing differentiation into cardiomyocytes using G-CSF

• US 8,252,583 B2
• Filed: 06/06/2008
• Issued: 08/28/2012
• Est. Priority Date: 06/07/2007
• Status: Active Grant

• Current Assignee: Keiichi Fukuda
• Original Assignee: Keiichi Fukuda
• Inventors: Fukuda, Keiichi, Yuasa, Shinsuke, Shimoji, Kenichiro
• Primary Examiner(s): Qian, Celine
• Assistant Examiner(s): Ault, Addison D
• Application Number:
  US12/663,464
• Publication Number:
  US 20110129922A1
• Time in Patent Office: 1,544 Days
• Field of Search: 435/477
• US Class Current: 435/325
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells

Abstract

A method for inducing differentiation of ES cells into cardiomyocytes, which comprises contacting the ES cells with an agonist for G-CSF receptor.

Claims

1. A method for inducing differentiation of ES cells into cardiomyocytes in vitro, which comprises culturing the ES cells in the presence of an agonist for G-CSF receptor, wherein the agonist for G-CSF receptor is added to the cell culture solution 5 to 8 days after removal of leukemia inhibitory factor (LIF) from the culture medium.

2. The method according to claim 1, wherein the agonist for G-CSF receptor is G-CSF.

3. The method according to claim 2, which comprises the following steps:

(a) culturing the ES cells;

(b) adding the G-CSF to a cell culture solution; and

(c) culturing the ES cells using the culture solution from step (b).

4. The method according to claim 1, which further comprises the step of culturing the ES cells in the presence of a substance that inhibits BMP signaling, prior to culturing the ES cells in the presence of the agonist for G-CSF receptor.

5. The method according to claim 4, wherein the substance that inhibits BMP signaling is Noggin.

11. The method according to claim 4, wherein the agonist for G-CSF receptor is G-CSF.

6. A method for producing cardiomyocytes in vitro, which comprises culturing ES cells in the presence of an agonist for G-CSF receptor, wherein the agonist for G-CSF receptor is added to the cell culture solution 5 to 8 days after removal of leukemia inhibitory factor (LIF) from the culture medium.

7. The method according to claim 6, wherein the agonist for G-CSF receptor is G-CSF.

8. The method according to claim 6, which comprises the following steps:

(a) culturing the ES cells;

(b) adding the G-CSF to the cell culture solution at the 5^th to 8^th day after removal of LIF from the culture medium; and

(c) culturing the ES cells using the culture solution from step (b).

9. The method according to claim 6, which further comprises the step of culturing the ES cells in the presence of a substance that inhibits BMP signaling, prior to culturing the ES cells in the presence of the agonist for G-CSF receptor.

10. The method according to claim 9, wherein the substance that inhibits BMP signaling is Noggin.