Method for producing anterior eye segment tissue

Patent metadata

Field	Value
Publication number	US-2016186136-A1
Application number	US-201414910332-A
Country	US
Kind code	A1
Filing date	Aug 6, 2014
Priority date	Aug 6, 2013
Publication date	Jun 30, 2016
Grant date	—

Abstract

The present invention provides a production method of a cell aggregate containing an anterior eye segment tissue or a partial structure thereof, or a precursor tissue thereof, including culturing pluripotent stem cell aggregates in suspension in the presence of a bone morphogenic factor signal transduction pathway activating substance to induce self-organization of an anterior eye segment tissue or a partial structure thereof, or a precursor tissue thereof. In one embodiment, the bone morphogenic factor signal transduction pathway activating substance is BMP4. In one embodiment, the suspension culture is performed entirely or partially in the presence of a fibroblast growth factor. The produced cell aggregate can further contain a neural retinal tissue.

First claim

1 . A production method of a cell aggregate comprising an anterior eye segment tissue or a partial structure thereof, or a precursor tissue thereof, comprising culturing an aggregate of pluripotent stem cells in suspension in the presence of a bone morphogenic factor signal transduction pathway activating substance.

2 . The method according to claim 1 , wherein the aggregate of pluripotent stem cells is cultured in suspension in the absence of a bone morphogenic factor signal transduction pathway activating substance, prior to the suspension culture in the presence of a bone morphogenic factor signal transduction pathway activating substance.

3 . The method according to claim 1 , wherein the bone morphogenic factor signal transduction pathway activating substance is BMP4

4 . The method according to claim 3 , wherein BMP4 has a concentration of 1-5 nM.

5 . The production method according to claim 1 , wherein the suspension culture is performed entirely or partially in the presence of a fibroblast growth factor.

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

7 . The method according to claim 1 , wherein the pluripotent stem cells are derived from human.

8 . The method according to claim 1 , wherein the suspension culture is performed in the absence of a feeder cell.

9 . The method according to claim 1 , wherein the cell aggregate further comprises a neural retinal tissue.

10 . The method according to claim 1 , wherein the anterior eye segment tissue is cornea and/or lens.

11 . The method according to claim 1 , wherein the cell aggregate comprises corneal epithelium as a partial structure of the anterior eye segment tissue, and further comprises a mesenchymal tissue, or corneal stroma and/or corneal endothelium derived therefrom.

12 . The method according to claim 11 , wherein the corneal epithelium is stratified.

13 . The method according to claim 1 , further comprising separating the anterior eye segment tissue or partial structure thereof or precursor tissue thereof from the cell aggregate.

14 . The method according to claim 13 , wherein the anterior eye segment tissue or partial structure thereof, or precursor tissue thereof is separated together with a neural retinal tissue.

15 . A cell aggregate obtained by the method according to claim 1 .

16 . An anterior eye segment tissue or a partial structure thereof, or a precursor tissue thereof, which is obtained by the method according to claim 13 .

Assignees

• Riken

Inventors

• Sasai Yoshiki
• Sasai Miyuki
• Ozone Chikafumi
• Maruyama Yuko

Classifications

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title

• C12N5/0621 Primary

  Eye cells, e.g. cornea, iris pigmented cells (photoreceptors C12N5/062) · CPC title

• C12N2506/02

  from embryonic cells · CPC title

• C12N2501/155

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

• C12N2500/90

  Serum-free medium, which may still contain naturally-sourced components · CPC title