US 12,227,725 B2
Cell culturing method and device
Ryo Suenaga, Yokohama (JP); and Maki Mitsuishi, Yokohama (JP)
Assigned to TOYO SEIKAN GROUP HOLDINGS, LTD., Tokyo (JP)
Appl. No. 16/960,816
Filed by TOYO SEIKAN GROUP HOLDINGS, LTD., Tokyo (JP)
PCT Filed Jan. 4, 2019, PCT No. PCT/JP2019/000011
§ 371(c)(1), (2) Date Jul. 8, 2020,
PCT Pub. No. WO2019/138956, PCT Pub. Date Jul. 18, 2019.
Claims priority of application No. 2018-001091 (JP), filed on Jan. 9, 2018.
Prior Publication US 2020/0354662 A1, Nov. 12, 2020
Int. Cl. C12N 5/00 (2006.01); C12M 1/00 (2006.01); C12M 3/06 (2006.01)
CPC C12M 23/14 (2013.01) [C12M 23/50 (2013.01); C12M 27/16 (2013.01); C12M 29/26 (2013.01); C12N 5/0068 (2013.01); C12N 2513/00 (2013.01); C12N 2527/00 (2013.01)] 8 Claims
 
1. A cell culturing method using a cell culture bag comprising a bag body configured from upper and lower surface films having a sealed periphery, and a port attached to the bag body, wherein a plurality of recesses are formed in the lower surface film, and wherein cells or cell aggregates are cultured in the plurality of recesses, comprising:
a closing-off step comprising pressing the upper surface film from above by means of a pressing member having a flat bottom surface, wherein during the pressing a culture medium contained in the bag body is discharged through the port such that the plurality of recesses are closed off by the upper surface film to confine the cells or the cell aggregates being cultured in each recess wherein the upper surface film is not in contact with the cells or cell aggregates;
a releasing step comprising releasing, in some or all of the plurality of recesses closed-off by the upper surface film, the cells or the cell aggregates adhering to inner surfaces of the recesses from inner surfaces of the recesses to change adhesion positions of the cells or the cell aggregates in each recess; and
a further culturing step comprising culturing the cells or the cell aggregates released from the inner surfaces of the recesses in each recess.