EP-4737555-A1 - LIQUID-FILLED CULTURE BAG, METHOD FOR PRODUCING LIQUID-FILLED CULTURE BAG, AND METHOD FOR USING LIQUID-FILLED CULTURE BAG

Abstract

Provided is a liquid-filled culture bag capable of efficiently removing air bubbles accumulated in minute wells in the culture bag without requiring a lot of labor or dedicated equipment, and capable of reducing troublesome work and the risk of contamination at a production site. A liquid-filled culture bag in which a culture bag having a microstructure formed in at least a part of a culture portion is housed in an outer packaging portion, wherein a space capable of being depressurized is present between an outside of the culture portion and the outer packaging portion, and the culture bag is vacuum-packaged. It is preferable that a plurality of first protrusions be provided outside the culture portion. In addition, it is preferable that a plurality of second protrusions be provided in a region of the outer packaging portion facing the outside of the culture portion. Furthermore, it is preferable that a spacer portion that forms a space capable of being depressurized be provided between the outside of the culture portion and the outer packaging portion.

Inventors

• TANAKA SATOSHI

Assignees

• Toyo Seikan Group Holdings, Ltd.

Dates

Publication Date

20260506

Application Date

20240614

Claims (12)

1. A liquid-filled culture bag in which a culture bag having a microstructure formed in at least a part of a culture portion is housed in an outer packaging portion, wherein a space capable of being depressurized is present between an outside of the culture portion and the outer packaging portion, and the culture bag is vacuum-packaged.
2. The liquid-filled culture bag according to claim 1, wherein a plurality of first protrusions are provided outside the culture portion.
3. The liquid-filled culture bag according to claim 1 or 2, wherein a plurality of second protrusions are provided in a region of the outer packaging portion facing the outside of the culture portion.
4. The liquid-filled culture bag according to claim 1 or 2, comprising a spacer portion that forms a space capable of being depressurized between the outside of the culture portion and the outer packaging portion.
5. The liquid-filled culture bag according to claim 1 or 2, wherein the microstructure is a plurality of wells or a plurality of grooves.
6. The liquid-filled culture bag according to claim 1 or 2, wherein an oxygen permeability of a member in a region provided with the culture portion in the culture bag is 3000 ml/m 2 ·day·atm or more.
7. The liquid-filled culture bag according to claim 1 or 2, wherein an oxygen permeability of a member in the region of the outer packaging portion facing the outside of the culture portion is equal to or less than the oxygen permeability of the culture portion.
8. The liquid-filled culture bag according to claim 7, wherein the member in the region of the outer packaging portion facing the outside of the culture portion has a gas barrier property with an oxygen permeability of 1000 ml/m 2 ·day·atm or less.
9. A method for producing a liquid-filled culture bag in which a culture bag having a microstructure formed in at least a part of a culture portion is housed in an outer packaging portion, a space capable of being depressurized being present between an outside of the culture portion and the outer packaging portion, the method comprising: filling the culture bag with a liquid; and vacuum-packaging the culture bag.
10. The method for producing a liquid-filled culture bag according to claim 9, wherein a plurality of first protrusions are provided outside the culture portion, and/or a plurality of second protrusions are provided in a region of the outer packaging portion facing the outside of the culture portion, and/or a spacer portion that forms a space capable of being depressurized is provided between the outside of the culture portion and the outer packaging portion.
11. A method for using the liquid-filled culture bag described in claim 1 or 2, the method comprising storing or transporting the liquid-filled culture bag at normal temperature, under refrigeration, or in a frozen state.
12. A method for using the liquid-filled culture bag according to claim 1 or 2, the method comprising injecting a cell, a culture medium, and cytokines, such as a cell adhesion factor and a cell growth factor, into the liquid-filled culture bag to perform culture.

Description

Technical Field The present invention relates to a cell culture technique, and particularly to a culture bag provided with minute wells or the like in a culture portion. Background Art When spheroids or organoids of adherent cells, such as iPS cells or ES cells, are cultured, a culture bag or the like, in which a large number of minute wells are formed in a culture portion, may be used. When such a culture bag is filled with a culture medium, air bubbles are trapped and accumulated in the minute wells, which causes a problem that interferes with cell culture. That is, when air bubbles remain in the minute wells of the culture bag, there is a problem that the formation of spheres (cell aggregates) composed of iPS cells and the like is inhibited, but it is extremely difficult to remove all the air bubbles from the minute wells. Citation List Patent Document Patent Document 1: WO 2021/241665 APatent Document 2: JP 2020-80670 A Summary of Invention Technical Problem As a means for removing the air bubbles accumulated in the minute wells, there is a method for removing the air bubbles by water pressure by spraying a liquid onto the air bubbles using, for example, a micropipette. In such an operation using a micropipette, however, there is a case where the air bubbles cannot be sufficiently removed particularly when the wells are small. In addition, this method is applicable only to an open container, and is not suitable for removing the air bubbles accumulated in the culture portion of the culture bag. Here, Patent Document 1 discloses that a culture bag having minute wells in a culture portion is filled with a culture medium, and then pressure is applied to the culture bag to remove air from the inside of the culture bag. In addition, Patent Document 2 discloses that air bubbles accumulated in minute wells are removed by filling a culture bag having minute wells in a culture portion with a degassed culture medium filling liquid. Furthermore, this document also discloses that air is removed from the inside of a culture bag by disposing the culture bag having minute wells in a culture portion in a vacuum exhaust device and exposing the culture bag to a vacuum. However, these methods have a problem that it takes a lot of labor to remove air bubbles and a problem that dedicated equipment is required. In addition, in the regenerative medicine and the production of cellular pharmaceuticals, an adjustment operation for removing air bubbles from a culture bag is required on site, and thus there is a problem in that the production process or the like becomes complicated and contamination may occur. Therefore, the present inventors have made intensive studies and developed a liquid-filled culture bag capable of solving these problems, thereby completing the present invention. That is, the present invention has been made in view of the above circumstances, and an object thereof is to provide a liquid-filled culture bag, a method for producing a liquid-filled culture bag, and a method for using a liquid-filled culture bag, which can efficiently remove air bubbles accumulated in minute wells in the culture bag without requiring a lot of labor or dedicated equipment, and can reduce troublesome work and the risk of contamination at a production site. Solution to Problem In order to achieve the above object, a liquid-filled culture bag of the present invention is a liquid-filled culture bag in which a culture bag having a microstructure formed in at least a part of a culture portion is housed in an outer packaging portion, and has a configuration in which a space capable of being depressurized is present between the outside of the culture portion and the outer packaging portion, and the culture bag is vacuum-packaged. In addition, the liquid-filled culture bag of the present invention is preferably configured to include a plurality of first protrusions outside the culture portion, is also preferably configured to include a plurality of second protrusions in a region of the outer packaging portion facing the outside of the culture portion, and is also preferably configured to include a spacer portion that forms a space capable of being depressurized between the outside of the culture portion and the outer packaging portion. In addition, it is preferable that the liquid-filled culture bag of the present invention has a configuration in which the microstructure is a plurality of wells or a plurality of grooves. In addition, it is preferable that the liquid-filled culture bag of the present invention has a configuration in which the oxygen permeability of a member in a region provided with the culture portion in the culture bag is 3000 ml/m2·day·atm or more. In addition, the liquid-filled culture bag of the present invention is preferably configured such that a member in the region of the outer packaging portion facing the outside of the culture portion has an oxygen permeability equal to or less than the oxygen permeability of th