IP Library › Granted Patent US 12,680,080
Granted Patent B2
US 12,680,080 · App. 17/806,398 · Granted Jul 14, 2026

Proliferative liver organoid, metabolically activated liver organoid, and use thereof

Inventors: Norio Masuda (Tokyo, JP); Toshiro Sato (Tokyo, JP); Ryo Igarashi (Tokyo, JP)
Assignees: JSR Corporation; KEIO UNIVERSITY
C12N5/0672C12N5/0606C12N5/0671C12N5/0677C12N5/0679C12N2501/115C12N2501/117C12N2501/12C12N2501/155C12N2501/16C12N2501/39C12N2501/999C12N2503/04C12N2506/02C12N2506/45C12N2510/00C12N2511/00C12N2513/00C12N2533/90
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Quick Facts
Patent No.
US 12,680,080
App. No.
17/806,398
Filed
Jun 10, 2022
Granted
Jul 14, 2026
Kind
B2
Art Unit
1632
USPC
435/171
Abstract

A production method for a proliferative liver organoid includes culturing liver stem cells or a tissue fragment including liver stem cells in a growth medium to obtain a proliferative liver organoid, in which the growth medium contains an interleukin-6 family cytokine. A production method for a metabolically activated liver organoid includes culturing the proliferative liver organoid produced by the production method for a proliferative liver organoid in a differentiation medium to obtain a metabolically activated liver organoid, in which the differentiation medium does not substantially contain an interleukin-6 family cytokine.

Claims (38)

1 . A method for producing a metabolically activated liver organoid, the method comprising:

culturing primary human hepatocytes or cryopreserved primary human hepatocytes in a growth medium and with contact to an extracellular matrix to obtain a proliferative liver organoid for at least 10 days and up to 14 days; and

culturing the proliferative liver organoid in a differentiation medium for 1 week to obtain a metabolically activated liver organoid,

wherein the growth medium comprises an interleukin-6 family cytokine, at least one growth factor, a Wnt agonist, a transforming growth factor-β inhibitor, a bone morphogenetic protein inhibitor, and a forskolin,

wherein the interleukin-6 family cytokine comprises at least one selected from the group consisting of interleukin-6, interleukin-11, oncostatin M, a leukemia inhibitory factor, cardiotrophin-1, and a ciliary neurotrophic factor,

wherein the differentiation medium comprises a growth factor, a transforming growth factor-β inhibitor, and a forskolin, and the differentiation medium does not substantially contain an interleukin-6 family cytokine,

wherein the extracellular matrix comprises basement membrane matrix, collagen I, or a 1:1 mixture of collagen I and basement membrane matrix, and

wherein the at least one growth factor in the growth medium is selected from the group consisting of:

epidermal growth factor;

hepatocyte growth factor;

heparin-binding EGF-like growth factor;

amphiregulin;

a combination of epidermal growth factor, fibroblast growth factor 10, and hepatocyte growth factor;

a combination of epidermal growth factor and fibroblast growth factor 10;

a combination of hepatocyte growth factor and fibroblast growth factor 10; and

a combination of epidermal growth factor and hepatocyte growth factor.

2 . The method according to claim 1 ,

wherein the growth medium does not substantially contain nicotinamide.

3 . The method according to claim 1 ,

wherein the growth factor in the growth medium further comprises at least one selected from the group consisting of an epidermal growth factor, a fibroblast growth factor, a hepatocyte growth factor, amphiregulin, and a heparin-binding EGF-like growth factor.

4 . The method according to claim 1 ,

wherein the growth medium further comprises a Rho kinase inhibitor.

5 . The method according to claim 1 ,

wherein the differentiation medium does not substantially contain nicotinamide.

6 . The method according to claim 1 ,

wherein the growth factor in the differentiation medium comprises at least one selected from the group consisting of an epidermal growth factor, a fibroblast growth factor, and a hepatocyte growth factor.

7 . The method according to claim 1 ,

wherein the differentiation medium further comprises a Wnt agonist.

8 . The method according to claim 1 ,

wherein the differentiation medium further comprises a Rho kinase inhibitor.

9 . The method according to claim 1 ,

wherein the differentiation medium further comprises a bone morphogenetic protein inhibitor.

10 . The method according to claim 1 ,

wherein the differentiation medium further comprises vitamin D.

11 . The method according to claim 1 ,

wherein the differentiation medium further comprises a Notch inhibitor.

12 . The method according to claim 1 , wherein the extracellular matrix comprises collagen I.

13 . The method according to claim 1 , wherein the extracellular matrix comprises laminin, entactin, and collagen IV.

Assignments (2)
MERGER AND CHANGE OF NAME Recorded Mar 31, 2025
From: JSR CORPORATION; JICC-02 CO., LTD.
To: JSR CORPORATION
Reel/Frame 070675/0565 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 10, 2022
From: MASUDA, NORIO; SATO, TOSHIRO; IGARASHI, RYO
To: JSR CORPORATION; KEIO UNIVERSITY
Reel/Frame 060170/0801 →
Priority Claims (1)
JP 2019-226717 · Dec 16, 2019 · national
Continuity (2)
Continuation PCTJP2020046781 · Dec 15, 2020
Related Publication 20220298485A1 · Sep 22, 2022
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