US 12,065,671 B2
Method for reproducible differentiation of clinical-grade retinal pigment epithelium cells
Kapil Bharti, Potomac, MD (US); Lucas Chase, Madison, WI (US); Feng Xuezhu, Madison, WI (US); and Balendu Shekhar Jha, Bethesda, MD (US)
Assigned to The United States of America, as represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US); and FUJIFILM Cellular Dynamics, Inc., Madison, WI (US)
Filed by The United States of America, as represented by the Secretary, Department of Health and Human Services, Bethesda, MD (US); and FUJIFILM Cellular Dynamics, Inc., Madison, WI (US)
Filed on Jul. 16, 2020, as Appl. No. 16/931,003.
Application 16/931,003 is a continuation of application No. 15/758,314, abandoned, previously published as PCT/US2016/050543, filed on Sep. 7, 2016.
Claims priority of provisional application 62/215,579, filed on Sep. 8, 2015.
Prior Publication US 2021/0139847 A1, May 13, 2021
Int. Cl. C12N 5/079 (2010.01)
CPC C12N 5/0621 (2013.01) [C12N 2500/30 (2013.01); C12N 2501/105 (2013.01); C12N 2501/15 (2013.01); C12N 2501/155 (2013.01); C12N 2501/16 (2013.01); C12N 2501/415 (2013.01); C12N 2506/45 (2013.01); C12N 2533/50 (2013.01); C12N 2533/52 (2013.01)] 30 Claims
 
1. A method for producing human retinal pigment epithelial (RPE) cells, comprising:
a) obtaining a starting population comprising human induced pluripotent stem cells (iPSCs) that are dissociated into essentially single cells;
b) culturing the iPSCs in a retinal induction medium at an initial cell density of about 1,000 to 33,000 cells/cm2 to initiate differentiation of the cells into retinal lineage cells;
c) further culturing the retinal lineage cells in a retinal differentiation medium to further differentiate the retinal lineage cells;
d) culturing the retinal lineage cells that were further differentiated in step (c) in a retinal medium to form differentiating RPE cells; and
e) culturing the differentiating RPE cells in a RPE maturation medium,
thereby producing RPE cells; wherein the method does not comprise the formation of embryoid bodies.