	US 11,697,794 B2
	Method for producing layered cell sheet and layered cell sheet produced by the same
Yuki Kagawa, Tokorozawa (JP); Tatsuya Shimizu, Tokyo (JP); Yuji Haraguchi, Tokyo (JP); and Hirotsugu Kubo, Tokorozawa (JP)
Assigned to NIHON KOHDEN CORPORATION, Tokyo (JP); and TOKYO WOMEN'S MEDICAL UNIVERSITY, Tokyo (JP)
Appl. No. 16/627,156
Filed by NIHON KOHDEN CORPORATION, Tokyo (JP); and TOKYO WOMEN'S MEDICAL UNIVERSITY, Tokyo (JP)
PCT Filed Jun. 28, 2018, PCT No. PCT/JP2018/024605 § 371(c)(1), (2) Date Dec. 27, 2019, PCT Pub. No. WO2019/004361, PCT Pub. Date Jan. 3, 2019.
Claims priority of application No. 2017-127873 (JP), filed on Jun. 29, 2017.
Prior Publication US 2020/0140807 A1, May 7, 2020
Int. Cl. C12N 5/00 (2006.01); C12N 5/077 (2010.01)

CPC C12N 5/0062 (2013.01) [C12N 5/0658 (2013.01); C12N 2513/00 (2013.01); C12N 2523/00 (2013.01)]

12 Claims

1. A method for producing a layered cell sheet on a temperature-responsive culture surface, the method comprising:

(1) a step of placing a first cell sheet on the temperature-responsive culture surface and applying a centrifugal force to the first cell sheet for a predetermined time in a temperature range from a lower critical solution temperature of the temperature-responsive culture surface to 45° C.,

(2) a step of further placing a second cell sheet on the first cell sheet on the temperature-responsive culture surface, and

(3) a step of applying a centrifugal force to the first cell sheet and the second cell sheet on the temperature-responsive culture surface for a predetermined time in the temperature range from the lower critical solution temperature to 45° C.,

wherein the centrifugal force is applied by a centrifuge having a heating function.