Cardiac organoid, manufacturing method therefor, and method for evaluating drug toxicity by using same

Patent metadata

Field	Value
Publication number	US-12486495-B2
Application number	US-202117906679-A
Country	US
Kind code	B2
Filing date	Feb 17, 2021
Priority date	Mar 19, 2020
Publication date	Dec 2, 2025
Grant date	Dec 2, 2025

Abstract

The present specification provides a spontaneous-contracting cardiac organoid, a method for manufacturing the organoid, and a method for evaluating drug toxicity by using same, the cardiac organoid comprising: a chamber in which a fluid is stored; a first pipe connected to the chamber so that the fluid flows therethrough; a second pipe connected to the chamber so that the fluid is discharged therethrough; and a valve formed on the first pipe so as to spontaneously open/close an inflow pipe.

First claim

The invention claimed is:

1 . A manufacturing method of a cardiac organoid, the method comprising:
a first culturing step of differentiating pluripotent stem cells (PSCs) into cardiomyocytes (CMs); a second culturing step of culturing the cardiomyocytes and an extracellular matrix (ECM) in a first maintenance medium so as to form a fusion tissue; and a third culturing step of culturing the fusion tissue in a second maintenance medium so as to form a cardiac organoid.

2 . The manufacturing method of the cardiac organoid of claim 1 , wherein the second maintenance medium contains insulin.

3 . The manufacturing method of the cardiac organoid of claim 1 , wherein the first culturing step comprises
seeding pluripotent stem cells; maintaining the seeded pluripotent stem cells in the first maintenance medium; culturing the pluripotent stem cells in an induction medium so that the pluripotent stem cells are induced into cardiac progenitors through a mesoderm cell stage; and culturing the mesoderm cells in the first maintenance medium to differentiate the cardiac progenitors into mature cardiomyocytes.

4 . The manufacturing method of the cardiac organoid of claim 3 , wherein the induction medium includes at least one of the group consisting of IWR-1 endo, XAV-939, JW74, SEN461, ICG-001, LGK-974, IWP-2, IWP-4, Wnt-C59 and WIKI4.

5 . The manufacturing method of the cardiac organoid of claim 3 , wherein the culturing of the pluripotent stem cells in the induction medium is performed for at least one period of 5 to 7 days.

6 . The manufacturing method of the cardiac organoid of claim 3 , wherein the culturing of the cardiac progenitors in the first maintenance medium is performed for at least one period of 10 to 21 days.

7 . The manufacturing method of the cardiac organoid of claim 1 , wherein the first maintenance medium does not contain insulin.

8 . The manufacturing method of the cardiac organoid of claim 1 , wherein the extracellular matrix is obtained from fibroblast.

9 . The manufacturing method of the cardiac organoid of claim 1 , wherein the second culturing step is performed for at least one period of 28 to 32 days.

10 . The manufacturing method of the cardiac organoid of claim 1 , wherein the third culturing step comprises
cutting the fusion tissue, and suspension-culturing the cut fusion tissue.

11 . A spontaneous-contracting cardiac organoid comprising:
a chamber in which a fluid is stored; a first pipe (track) connected to the chamber so that the fluid flows therethrough; a second pipe connected to the chamber so that the fluid is discharged therethrough; and a valve formed on the first pipe to spontaneously open/close an inflow pipe.

12 . The spontaneous-contracting cardiac organoid of claim 11 , wherein the chamber expresses TUBB3, TNNT2, PECAM1 and MYL2.

13 . The spontaneous-contracting cardiac organoid of claim 11 , wherein in the chamber, trabeculated cardiomyocytes are formed toward an inner pipe of the chamber.

14 . The spontaneous-contracting cardiac organoid of claim 11 , wherein the chamber is formed with calcium transients.

15 . A method for evaluating drug toxicity by using a cardiac organoid, the method comprising:
reacting the cardiac organoid of claim 11 with a drug; washing the cardiac organoid after the drug reaction is completed; culturing the washed cardiac organoid; capturing images of the reacting, washing and culturing; obtaining the captured images; and analyzing the obtained images.

16 . The method of claim 15 , wherein the analyzing of the images is performed based on a difference in amount of change of pixel values between a cell area and a background area in images continuously captured during contraction of the cardiac organoid.

17 . The method of claim 15 , wherein the analyzing of the images is performed by measuring conduction displacement, beat rate variation, and beating velocity.

Assignees

• Univ Yonsei Iacf

Inventors

• Yoon Young Sup
• Cho Jae Yeaon

Classifications

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• C12N2513/00

  3D culture · CPC title

• C12N2506/45

  from artificially induced pluripotent stem cells · CPC title

• C12N2501/415

  Wnt; Frizzeled · CPC title

• C12N2501/16

  Activin; Inhibin; Mullerian inhibiting substance · CPC title