Generation of Thymic Epithelial Progenitor Cells In Vitro

Patent metadata

Field	Value
Publication number	US-2016010055-A1
Application number	US-201414770625-A
Country	US
Kind code	A1
Filing date	Feb 26, 2014
Priority date	Feb 27, 2013
Publication date	Jan 14, 2016
Grant date	—

Abstract

Methods for generating thymic epithelial progenitor (TEP) cells from pluripotent stem (PS) cells in vitro are provided. Compositions and systems of cell populations of TEP cells as well as cells formed during different stages of differentiation of PS cells into TEP cells are also disclosed. The methods, isolated in vitro cell populations, compositions, and systems disclosed provide functional TEP cells that mature into thymic epithelial cells in vivo.

First claim

What is claimed is:

1 . A method for generating thymic epithelial progenitor (TEP) cells, the method comprising:
culturing definitive endodermal (DE) cells obtained from pluripotent stem cells in a medium comprising an activator of retinoic acid receptor, an activator of bone morphogenetic protein (BMP) signaling, and an inhibitor of transforming growth factor-β (TGF-β) signaling.

2 . The method of claim 1 , wherein the DE cells are obtained from pluripotent stem cells by culturing pluripotent stem cells in a medium comprising a growth factor selected from the group consisting of Nodal, Activin A, and Activin B.

3 . The method of claim 1 or 2 , wherein the method comprises:
culturing anterior foregut endodermal (AFE) cells produced by said culturing of the DE cells, wherein said culturing of the AFE cells is in a medium comprising an activator of retinoic acid receptor, an activator of bone morphogenetic protein (BMP) signaling, and an inhibitor of transforming growth factor-β (TGF-β) signaling.

4 . The method of claim 3 , wherein the method comprises:
culturing ventral pharyngeal endodermal (VPE) cells produced by said culturing of the AFE cells, wherein said culturing of the VPE cells is in a medium comprising an activator of retinoic acid receptor and an activator of bone morphogenetic protein (BMP) signaling.

5 . A method for generating thymic epithelial progenitor (TEP) cells, the method comprising:
culturing anterior foregut endodermal (AFE) cells obtained from pluripotent stem cells in a medium comprising an activator of retinoic acid receptor, an activator of bone morphogenetic protein (BMP) signaling, and an inhibitor of transforming growth factor-β (TGF-β) signaling.

6 . The method of claim 5 , wherein the method comprises:
culturing ventral pharyngeal endodermal (VPE) cells produced by said culturing of the AFE cells, wherein said culturing of the VPE cells is in a medium comprising an activator of retinoic acid receptor and an activator of bone morphogenetic protein (BMP) signaling.

7 . A method for generating thymic epithelial progenitor (TEP) cells, the method comprising:
culturing ventral pharyngeal endodermal (VPE) cells obtained from pluripotent stem cells in a medium comprising an activator of retinoic acid receptor and an activator of bone morphogenetic protein (BMP) signaling.

8 . The method of any one of claims 1 - 7 , wherein the pluripotent stem cells are selected from the group consisting of embryonic stem cell, embryonic germ cells, and induced pluripotent stem cell.

9 . The method of claim 8 , wherein the pluripotent stem cells are primate pluripotent stem cells (pPS) cells.

10 . The method of claim 9 , wherein the pPS cells are human pluripotent stem (hPS) cells.

11 . The method of claim 10 , wherein the hPS cells are human embryonic stem (hES) cells.

12 . The method of claim 10 , wherein the hPS cells are induced pluripotent stem (iPS) cells.

13 . A composition comprising:
thymic epithelial progenitor (TEP) cells; an activator of retinoic acid receptor; and an activator of BMP signaling.

14 . The composition of claim 13 , further comprising:
a Wnt family member; a fibroblast growth factor; and an inhibitor of hedgehog signaling.

15 . A composition comprising:
definitive endodermal (DE) cells; an activator of retinoic acid receptor; an activator of bone morphogenetic protein (BMP) signaling; and an inhibitor of transforming growth factor-β (TGF-β) signaling.

16 . A composition comprising:
anterior foregut endodermal (AFE) cells; an activator of retinoic acid receptor; an activator of bone morphogenetic protein (BMP) signaling; and an inhibitor of transforming growth factor-β (TGF-β) signaling.

17 . The composition of claim 16 , further comprising:
Wnt family member; a fibroblast growth factor; and an inhibitor of hedgehog signaling.

18 . A composition comprising:
ventral pharyngeal endodermal (VPE) cells; an activator of retinoic acid receptor; and an activator of bone morphogenetic protein (BMP) signaling.

19 . The composition of claim 18 , further comprising:
Wnt family member; a fibroblast growth factor; and an inhibitor of hedgehog signaling.

20 . A first in vitro cell population including primate cells and a second in vitro cell population comprising progeny of a portion of the first in vitro cell population, wherein the progeny are TEP cells.

21 . The first and second cell populations of claim 20 , wherein the TEP cells express FOXN1.

22 . The first and second cell populations of claims 20 - 21 , wherein the first in vitro cell population comprises primate pluripotent stem cells.

23 . The first and second cell populations of claims 20 - 21 , wherein the first in vitro cell population comprises DE cells.

24 . The first and second cell populations of claims 20 - 21 , wherein the first in vitro cell population comprises AFE cells.

25 . The first and second cell populations of claims 20 - 21 , wherein the first in vitro cell population comprises VPE cells.

26 . A first in vitro cell population including primate pluripotent stem cells and a second in vitro cell population comprising progeny of a portion of the first in vitro cell population, wherein the progeny are DE cells.

27 . The first and second cell populations of claim 26 , wherein the DE cells express SOX17.

28 . A first in vitro cell population including primate pluripotent stem cells and a second in vitro cell population comprising progeny of a portion of the first in vitro cell population, wherein the progeny are AFE cells.

29 . A first in vitro cell population including primate pluripotent stem cells and a second in vitro cell population comprising progeny of a portion of the first in vitro cell population, wherein the progeny are VPE cells.

30 . A system for generating TEP cells, the system comprising:
a line of undifferentiated human PS cells; and a cell population of TEP cells differentiated therefrom, wherein the TEP cells express one or more of the TEP cell markers.

31 . A system for generating TEP cells, the system comprising:
a cell population of human DE cells; and a cell population of TEP cells differentiated therefrom

32 . A system for generating TEP cells, the system comprising:
human AFE cells; and a cell population of TEP cells differentiated therefrom.

33 . A system for generating TEP cells, the system comprising:
human VPE cells; and a cell population of TEP cells differentiated therefrom.

34 . A system for generating TEP cells, the system comprising:
human PS cells and a cell population of DE cells differentiated therefrom.

35 . A system for generating TEP cells, the system comprising:
human PS cells; and a cell population of AFE cells differentiated therefrom.

36 . A system for generating TEP cells, the system comprising:
human PS cells and a cell population of VPE cells differentiated therefrom, wherein the VPE cells express one or more of the VPE cell markers

37 . A system for generating TEP cells, the system comprising:
human PS cells, a cell population of DE cells differentiated from the PS cells, a cell population of A

Assignees

• Univ California

Inventors

• Parent Audrey
• Hebrok Matthias
• Anderson Mark Stuart

Classifications

• C12N2506/45

  from artificially induced pluripotent stem cells · CPC title

• C12N2501/385

  of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR] · CPC title

• C12N2501/15

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

• C12N2501/119

  Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10 · CPC title

• C12N2501/155

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