US 12,378,520 B2
Method for producing retinal tissue and retina-related cells
Tokushige Nakano, Osaka (JP); Yoshiki Sasai, Kobe (JP); and Chikafumi Ozone, Wako (JP)
Assigned to SUMITOMO CHEMICAL COMPANY, LIMITED, Tokyo (JP); and RIKEN, Wako (JP)
Filed by SUMITOMO CHEMICAL COMPANY, LIMITED, Tokyo (JP); and RIKEN, Wako (JP)
Filed on Jul. 21, 2022, as Appl. No. 17/870,569.
Application 17/870,569 is a continuation of application No. 16/692,484, filed on Nov. 22, 2019, granted, now 11,473,056.
Application 16/692,484 is a continuation of application No. 14/913,628, granted, now 10,501,724, issued on Dec. 10, 2019, previously published as PCT/JP2014/072065, filed on Aug. 22, 2014.
Claims priority of application No. 2013-173285 (JP), filed on Aug. 23, 2013.
Prior Publication US 2022/0364054 A1, Nov. 17, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/079 (2010.01); A61K 35/30 (2015.01); A61L 27/38 (2006.01)
CPC C12N 5/0621 (2013.01) [A61K 35/30 (2013.01); A61L 27/3804 (2013.01); A61L 27/3834 (2013.01); A61L 27/3895 (2013.01); A61L 2430/16 (2013.01); C12N 2500/33 (2013.01); C12N 2500/44 (2013.01); C12N 2500/90 (2013.01); C12N 2501/155 (2013.01); C12N 2501/19 (2013.01); C12N 2501/385 (2013.01); C12N 2501/415 (2013.01); C12N 2501/727 (2013.01); C12N 2506/02 (2013.01); C12N 2506/45 (2013.01)] 21 Claims
 
1. A method for producing a cell population containing retinal progenitor cells, comprising
(1) a first step of subjecting pluripotent stem cells to floating culture in a serum-free medium to form an aggregate of pluripotent stem cells,
(2) a second step of subjecting the aggregate formed in step (1) to floating culture in a serum-free medium or serum-containing medium each being free of a substance acting on the Sonic hedgehog (Shh) signal transduction pathway that can enhance signal transduction mediated by Shh at a concentration exerting an adverse influence on the selective differentiation into retinal progenitor cell and retinal tissue and containing a substance acting on the bone morphogenic protein (BMP) signal transduction pathway that can enhance signal transduction pathway mediated by BMP at a concentration necessary for differentiation induction into retinal cells from day 1 or later from the start of the floating culture in step (1) until a cell expressing retina and anterior neural fold homeobox (Rax) gene appears, thereby obtaining an aggregate containing retinal progenitor cells, and
(3) a third step of dispersing the aggregate containing retinal progenitor cells formed in step (2), thereby obtaining a dispersed cell population containing retinal progenitor cells.