Cell collection method

Patent metadata

Field	Value
Publication number	US-9574168-B2
Application number	US-201414341543-A
Country	US
Kind code	B2
Filing date	Jul 25, 2014
Priority date	Feb 1, 2012
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

A method for collecting cells that have been cultured by using a cell culture kit in which, at least, a culture container for cultivating cells, a cell collection container for collecting cells after cultivation and a waste liquid container for collecting a culture medium after cultivation are linked to one another through a conduit is provided. The method includes transferring a culture supernatant that has been cultured from the culture container to the waste liquid container, transferring a concentrated suspension of cells from the culture container to the cell collection container, returning part of the culture supernatant discharged in the waste liquid container to the culture container and allowing cells remaining in the culture container to be suspended in a suspension of cells, and transferring again a suspension of cells in which remaining cells are suspended from the culture container to the cell collection container.

First claim

The invention claimed is:

1. A method for collecting cultured cells by using a cell culture kit comprising a culture container for cultivating cells, a cell collection container for collecting cells after cultivation and a waste liquid container for collecting a culture medium after cultivation, wherein the culture container, the cell collection container, and the waste liquid container are linked to one another through a conduit, the method comprising:
discharging a culture supernatant by transferring the culture supernatant from the culture container to the waste liquid container to obtain a concentrated suspension;
collecting cells by transferring the concentrated suspension of the cultured cells from the culture container to the cell collection container;
returning part of the culture supernatant that had been discharged into the waste liquid container to the culture container to allow cells remaining in the culture container to be suspended in a suspension of cells; and
re-collecting the cells remaining in the culture container by transferring the suspension of the cells from the culture container to the cell collection container.

2. The method according to claim 1 , wherein, in the returning, the cells remaining in the culture container are dispersed in the suspension of the cells while pressing a surface of the culture container until the surface of the culture container is lowered to a predetermined depth by a stirring element and moving the stirring element reciprocally in a horizontal direction on the culture container.

3. The method according to claim 2 , wherein the dispersing of the cells in the returning and the transferring of the suspension in the re-collecting are conducted simultaneously.

4. The method according to claim 1 , wherein the returning and the re-collecting are repeated twice or more.

5. The method according to claim 1 , wherein the waste liquid container is a culture medium storage container.

6. The method according to claim 1 , further comprising, prior to discharging a culture supernatant:
reducing an inner bottom surface area of the culture container, wherein the reducing is performed by moving a partition element on the culture container while pressing the culture container by the partition element from the end portion thereof to bring an upper surface and a lower surface thereof into close contact with each other, and
settling the cultured cells in the culture container by standing still after the reducing.

7. A method for collecting cultured cells by using a cell culture kit comprising a culture container for cultivating cells, a cell collection container for collecting cells after cultivation and a waste liquid container for collecting a culture medium after cultivation, wherein the culture container, the cell collection container, and the waste liquid container are linked to one another through a conduit, the method comprising:
discharging a culture supernatant by transferring the culture supernatant from the culture container to the waste liquid container to obtain a concentrated suspension;
reducing an inner bottom surface area of the culture container to gather the concentrated suspension of the cultured cells to a cell collection port side of the culture container, wherein the reducing is performed by moving a partition element on the culture container while pressing the culture container by the partition element from the outside of a cultivation space to bring an upper surface and a lower surface thereof into close contact with each other; and
collecting the gathered suspension by transferring the concentrated suspension from the culture container to the cell collection container.

8. The method according to claim 7 , wherein the gathering of the concentrated suspension of the cultured cells in the reducing and the transferring of the concentrated suspension in the collecting are conducted simultaneously.

9. The method according to claim 7 , further comprising collecting cells by transferring the concentrated suspension of the cultured cells from the culture container to the cell collection container, wherein the collecting is performed after the discharging and is followed by the reducing.

10. The method according to claim 7 , further comprising, after collecting the gathered suspension:
returning part of the culture supernatant that had been discharged into the waste liquid container to the culture container to allow cells remaining in the culture container to be suspended in a suspension of cells; and
re-collecting the cells remaining in the culture container by transferring the suspension of the cells from the culture container to the cell collection container.

11. The method according to claim 7 , wherein the partition element is a roller.

Assignees

• Toyo Seikan Group Holdings Ltd

Inventors

• Tanaka Satoshi
• Suenaga Ryo
• Kuninori Masahiro
• Ota Kyohei

Classifications

• C12M33/18

  Rollers · CPC title

• C12N1/02 Primary

  Separating microorganisms from their culture media · CPC title

• C12M23/14

  Bags · CPC title

• C12M3/00

  Tissue, human, animal or plant cell, or virus culture apparatus · CPC title

• C12M1/00

  Apparatus for enzymology or microbiology · CPC title