US 11,987,810 B2
RPE cell populations and methods of generating same
Osnat Bohana-Kashtan, Tel-Mond (IL); Lior Ann Rosenberg Belmaker, Shoham (IL); and Ofer Wiser, Jerusalem (IL)
Assigned to CELL CURE NEUROSCIENCES LTD., Jerusalem (IL)
Filed by Cell Cure Neurosciences Ltd., Jerusalem (IL)
Filed on Dec. 8, 2021, as Appl. No. 17/546,013.
Application 17/546,013 is a division of application No. 15/539,473, granted, now 11,891,622, previously published as PCT/IL2015/051269, filed on Dec. 30, 2015.
Claims priority of provisional application 62/195,309, filed on Jul. 22, 2015.
Claims priority of provisional application 62/116,972, filed on Feb. 17, 2015.
Claims priority of provisional application 62/097,753, filed on Dec. 30, 2014.
Prior Publication US 2022/0154141 A1, May 19, 2022
Int. Cl. C12N 5/079 (2010.01); A61K 35/30 (2015.01)
CPC C12N 5/0621 (2013.01) [A61K 35/30 (2013.01); C12N 2500/02 (2013.01); C12N 2500/38 (2013.01); C12N 2501/115 (2013.01); C12N 2501/15 (2013.01); C12N 2501/16 (2013.01); C12N 2506/02 (2013.01); C12N 2533/52 (2013.01)] 15 Claims
 
1. A method of generating retinal pigment epithelium (RPE) cells, comprising:
(a) culturing human pluripotent stem cells for about 1 week to about 3 weeks in a medium comprising nicotinamide so as to generate differentiating cells, wherein said medium is devoid of Activin A;
(b) further culturing said differentiating cells for about 1 week to about 4 weeks in a medium comprising Activin A and nicotinamide to generate cells which are further differentiated towards an RPE lineage;
(c) further culturing said cells which are further differentiated towards the RPE lineage for up to 4 weeks in a medium comprising nicotinamide so as to generate RPE cells, wherein said medium is devoid of Activin A,
wherein steps (a)-(c) are effected under conditions wherein the atmospheric oxygen level is less than about 10%, wherein at least 80% of the RPE cells co-express premelanosome protein (PM EL17) and cellular retinaldehyde binding protein (CRALBP) after step (c); and (d)
expanding said RPE cells.