Method of culturing cell, method of manufacturing cell support complex, cultured cell, and cell support complex

Patent metadata

Field	Value
Publication number	US-12221597-B2
Application number	US-202017006138-A
Country	US
Kind code	B2
Filing date	Aug 28, 2020
Priority date	Mar 22, 2018
Publication date	Feb 11, 2025
Grant date	Feb 11, 2025

Abstract

A method of culturing cells includes culturing the cell in a cell suspension under repetitive cycles of stirring and rest while keeping the cell non-adherent to a culture vessel, so as to form at least one aggregate of the cells.

First claim

What is claimed is:

1. A method of culturing cells comprising:
culturing the cells in a cell suspension under repetitive cycles of stirring and rest while keeping the cells non-adherent to a culture vessel, wherein the culture vessel is treated by cell-repellent treatment or composed of a cell-repellent material,
forming at least one aggregate of the cells without applying a support in the cell suspension; and
subsequent to the steps of forming at least one aggregate and culturing the cells under repetitive cycles of stirring and rest, culturing the cells in a state of aggregate, with the cell suspension kept stirred.

2. The method of culturing cells according to claim 1 , wherein rest time is longer than 5 minutes and shorter than 480 minutes.

3. The method of culturing cells according to claim 1 , wherein the culture vessel has a stirring blade for stirring the cell suspension, with a rotating speed exceeding 20 rpm.

4. The method of culturing cells according to claim 1 , wherein the stirring applies a shear stress of 0.3 N/m2 or smaller in absolute value to the cells.

5. The method of culturing cells according to claim 1 , wherein culture period of the cells is between 5 days and 14 days.

6. The method of culturing cells according to claim 1 , wherein the aggregate is composed of cells with numbers between 500 and 5000.

7. The method of culturing cells according to claim 1 , wherein size of the aggregate is between 100 μm and 350 μm.

8. The method of culturing cells according to claim 1 , wherein the cells are dedifferentiated cells, and restore the physiological functions of the cells by forming the aggregate.

9. The method of culturing cells according to claim 1 , wherein the rest time is between 10 minutes and 30 minutes.

10. The method of culturing cells according to claim 1 , wherein the cells are renal cells.

11. The method of culturing cells according to claim 1 , further comprising seeding the cells in the culture vessel with a medium and resting the cells for a predetermined duration of time,
wherein the step of culturing the cells in the cell suspension is performed after resting the cells in the medium.

Assignees

• Nikkiso Co Ltd

Inventors

• Kitagawa Fumihiko

Classifications

• C12M27/02

  Stirrer or mobile mixing elements · CPC title

• C12N1/00

  Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor · CPC title

• C12M23/20 Primary

  Material Coatings (immunocoatings C12M25/00) · CPC title