US 12,291,720 B2
Production method for retinal tissue
Atsushi Kuwahara, Kobe (JP); Suguru Yamasaki, Kobe (JP); Yasushi Hiramine, Kobe (JP); Yoshiki Sasai, Wako (JP); and Masayo Takahashi, Wako (JP)
Assigned to SUMITOMO PHARMA CO., LTD., Osaka (JP); and RIKEN, Wako (JP)
Filed by SUMITOMO PHARMA CO., LTD., Osaka (JP); and RIKEN, Wako (JP)
Filed on Jan. 3, 2022, as Appl. No. 17/567,708.
Application 17/567,708 is a continuation of application No. 15/521,387, granted, now 11,214,772, previously published as PCT/JP2015/080017, filed on Oct. 23, 2015.
Claims priority of application No. 2014-217868 (JP), filed on Oct. 24, 2014.
Prior Publication US 2022/0119764 A1, Apr. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/079 (2010.01); A61K 35/30 (2015.01); A61K 35/44 (2015.01); A61L 27/00 (2006.01); A61L 27/38 (2006.01); C12N 5/0793 (2010.01); C12N 5/0797 (2010.01); C12Q 1/02 (2006.01); G01N 33/50 (2006.01)
CPC C12N 5/0621 (2013.01) [A61K 35/30 (2013.01); A61K 35/44 (2013.01); A61L 27/00 (2013.01); A61L 27/383 (2013.01); A61L 27/3895 (2013.01); C12N 5/0619 (2013.01); C12N 5/0623 (2013.01); C12Q 1/02 (2013.01); G01N 33/5014 (2013.01); G01N 33/5058 (2013.01); C12N 2501/115 (2013.01); C12N 2501/155 (2013.01); C12N 2501/41 (2013.01); C12N 2501/50 (2013.01); C12N 2501/999 (2013.01); C12N 2506/45 (2013.01); C12N 2533/52 (2013.01)] 18 Claims
 
1. A method for producing a cell aggregate containing a mixture of cells expressing Oct3/4 and cells expressing at least one kind selected from the group consisting of SOX1, N-cadherin, OTX2 and Nestin, comprising the following steps (1) and (2):
(1) a first step of culturing human pluripotent stem cells in the absence of feeder cells and in a medium comprising a factor for maintaining undifferentiated state of the human pluripotent stem cells, and
(2) a second step of dispersing the cells obtained in the first step and culturing the dispersed cells in suspension in the presence of a Sonic hedgehog signal transduction pathway activating substance in the absence of a Wnt signal transduction pathway inhibiting substance to form a cell aggregate within about 72 hours after the cells obtained in the first step are dispersed.