IP Library › Granted Patent US 12,350,294
Granted Patent B2
US 12,350,294 · App. 16/962,590 · Granted Jul 8, 2025

Medium comprising serum replacement, IGF1, STAT3 activator and adrenal gland hormone without BFGF or rock inhibitor for producing corneal endothelial substitute cells from iPS cells and a method thereof

Inventors: Shigeto Shimmura (Tokyo, JP); Shin Hatou (Tokyo, JP); Kazuo Tsubota (Tokyo, JP)
Assignee: KEIO UNIVERSITY
A61K35/30A61P27/02C12N5/0621
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Quick Facts
Patent No.
US 12,350,294
App. No.
16/962,590
Granted
Jul 8, 2025
Kind
B2
Abstract

Using the medium of the present invention for inducing a cornea endothelial substitute cell from an iPS cell, the medium containing an insulin-like growth factor and a STAT3 activator, and not containing a basic fibroblast growth factor or a ROCK inhibitor, and the method of the present invention for inducing corneal endothelial substitute cells from iPS cells by using the medium, it becomes possible to efficiently produce corneal endothelial substitute cells, particularly to efficiently produce corneal endothelial substitute cells from iPS cells. Furthermore, it becomes possible to stably produce large amounts of corneal endothelial substitute cells by inducing differentiation of iPS cells into corneal endothelial substitute cells more efficiently.

Claims (36)

1. A medium for inducing a corneal endothelial substitute cell that expresses Na,K-ATPase al subunit (ATP1A1), ZO-1, N-cadherin, and PITX2 from a somatic cell derived induced pluripotent stem (iPS) cell that expresses Oct4, Lin28A, and Sox9, the medium comprising

a basal medium selected from the group consisting of MEM medium, BME medium, BGJb medium, CMRL 1066 medium, Glasgow MEM medium, Improved MEM Zinc Option medium, IMDM medium, Medium 199 medium, Eagle MEM medium, aMEM medium, DMEM medium, ham medium, RPMI 1640 medium, Fischer's medium, a mixed medium thereof, and DMEM/F12 medium,

a serum replacement,

insulin-like growth factor 1 (IGF1),

a signal transducer and activator of transcription 3 (STAT3) activator that is one or both of interleukin-6 (IL-6) and leukemia inhibitory factor (LIF),

adrenal gland hormone selected from the group consisting of dexamethasone, hydrocortisone, betamethasone, beclomethasone, aldosterone, dehydroepiandrosterone, and androstenedione, and

not comprising a basic fibroblast growth factor or a Rho-associated, coiled-coil-containing kinase (ROCK) inhibitor.

2. The medium according to claim 1 , further comprising a keratinocyte growth factor.

3. The medium according to claim 1 , further comprising transforming growth factor beta 1 (TGF-beta 1).

4. The medium according to claim 1 , further comprising ascorbic acid.

5. The medium according to claim 1 , wherein the basal medium is DMEM/F12 medium.

6. The medium according to claim 1 , wherein the adrenal gland hormone is selected from the group consisting of aldosterone, dexamethasone, and hydrocortisone.

7. The medium according to claim 1 , which is a medium comprising

DMEM/F12 medium as a basal medium,

insulin-like growth factor 1 (IGF1),

a signal transducer and activator of transcription 3 (STAT3) activator that is one or both of interleukin-6 (IL-6) and leukemia inhibitory factor (LIF),

dexamethasone, and

aldosterone.

8. A method for producing a corneal endothelial substitute cell that expresses Na,K-ATPase al subunit (ATP1A1), ZO-1, N-cadherin, and PITX2 from a somatic cell derived induced pluripotent stem (iPS) cell that expresses Oct4, Lin28A, and Sox9, the method comprising adhesion-culturing in a medium comprising

a basal medium selected from the group consisting of MEM medium, BME medium, BGJb medium, CMRL 1066 medium, Glasgow MEM medium, Improved MEM Zinc Option medium, IMDM medium, Medium 199 medium, Eagle MEM medium, aMEM medium, DMEM medium, ham medium, RPMI 1640 medium, Fischer's medium, a mixed medium thereof, and DMEM/F12 medium,

a serum replacement,

insulin-like growth factor 1 (IGF1),

a signal transducer and activator of transcription 3 (STAT3) activator that is one or both of interleukin-6 (IL-6) and leukemia inhibitory factor (LIF),

adrenal gland hormone selected from the group consisting of dexamethasone, hydrocortisone, betamethasone, beclomethasone, aldosterone, dehydroepiandrosterone, and androstenedione, and

not comprising a basic fibroblast growth factor or a Rho-associated, coiled-coil-containing kinase (ROCK) inhibitor.

9. The method according to claim 8 , wherein the medium further comprises a keratinocyte growth factor.

10. The method according to claim 8 , wherein the medium further comprises transforming growth factor beta 1 (TGF-beta 1).

11. The method according to claim 8 , further comprising ascorbic acid.

12. The method according to claim 8 , wherein the basal medium is DMEM/F12 medium.

13. The method according to claim 8 , wherein the adrenal gland hormone is selected from the group consisting of aldosterone, dexamethasone, and hydrocortisone.

14. The medium according to claim 8 , which is a medium comprising

DMEM/F12 medium as a basal medium,

insulin-like growth factor 1 (IGF1),

a signal transducer and activator of transcription 3 (STAT3) activator that is one or both of interleukin-6 (IL-6) and leukemia inhibitory factor (LIF),

dexamethasone, and

aldosterone.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 4, 2020
From: SHIMMURA, SHIGETO; HATOU, SHIN; TSUBOTA, KAZUO
To: KEIO UNIVERSITY
Reel/Frame 053700/0441 →
Priority Claims (1)
JP 2018-005076 · Jan 16, 2018 · national
Continuity (1)
Related Publication 20200397826A1 · Dec 24, 2020
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