IP Library › Patent Application 16694040
Patent Application
App. No. 16/694,040

EFFICIENT INDUCTION OF DEFINITIVE ENDODERM FROM PLURIPOTENT STEM CELLS

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Quick Facts
Patent No.
US None
App. No.
16/694,040
Abstract

The present invention relates to a method to differentiate pluripotent stem cells to a primitive streak cell population, in a stepwise manner for further maturation to definitive endoderm.

Claims (19)

1 . A method for differentiation of stem cells into definitive endoderm comprising the steps of:

incubating stem cells in a medium comprising at least 2 μM CHIR, wherein activin A is not present; and

subsequently incubating stem cells in a medium comprising activin A.

2 . The method according to claim 1 , wherein said medium is RPMI-1640.

3 . The method according to claim 1 , wherein said stem cells are embryonic stem cells or induced pluripotent stem cells.

4 . The method according to claim 1 , wherein the concentration of CHIR is at least about 2.5 μM.

5 . The method according to claim 1 , wherein the concentration of CHIR is at least about 3.1 μM.

6 . The method according to claim 1 , wherein the concentration of CHIR is in a range selected from the group consisting of about 2.5-15 μM, about 3.1-15 μM, about 3.1-7 μM, 3.5-7 μM, about 3.5-6 μM, and about 3.5-5 μM.

7 . The method according to claim 1 , wherein the concentration of CHIR is at least about 3.5 μM.

8 . The method according to claim 1 , wherein said incubation with CHIR is at least 24 hours.

9 . The method according to claim 1 , wherein said incubation with activin A is at least 24 hours.

10 . The method according to claim 1 , wherein said incubation with activin A is 48 to 72 hours.

11 . The method according to claim 1 , wherein endodermal cells are obtained from said definitive endoderm cells.

12 . The method according to claim 11 , wherein said endoderm cells are pancreatic endoderm cells.

13 . Pancreatic endoderm cells obtainable by the method of claim 12 .

14 . The method according to claim 1 , wherein the concentration of CHIR in the culture medium is at least 3 μM.

15 . The method according to claim 1 , wherein the concentration of CHIR in the culture medium is in a range selected from the group consisting of about 2-15 μM, about 3-15 μM, about 3.1-15 μM, about 3.1-7 μM, about 3.5-15 μM, and about 3.5-7 μM.

16 . A method for inducing primitive streak cells from embryonic stem cells by using CHIR in a concentration of at least 2 μM in the culture medium.

17 . A method for inducing definitive endoderm cells from embryonic stem cells by using CHIR in a concentration in the range of 3.5-7 μM.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 1, 2026
From: NOVO NORDISK A/S
To: ASPECT BIOSYSTEMS LTD.
Reel/Frame 075363/0935 →