a) differentiating a population of human pluripotent stem cells in a medium comprising nicotinamide, in the absence of activin A, under non-adherent conditions to generate a cluster of cells comprising differentiating cells;

b) further differentiating said differentiating cells of (a) in a medium comprising nicotinamide, in the absence of activin A under adherent conditions;

c) further differentiating the population of differentiating cells in a culture comprising a medium comprising one or more members of the TGFβ superfamily, thereby generating a mixed population of cells comprising RPE cells;

d) enzymatically removing said mixed population of cells from said culture without mechanical isolation of cells, wherein more than 10% of the cells of said mixed populations of cells are non-pigmented cells; and

e) expanding said mixed population of cells to generate an expanded population of RPE cells.