Methods for producing retinal pigment epithelium cells

A method of enriching RPE cells or progenitors of RPE cells in a mixture of cells, comprising:

• a. labeling a cell surface marker in a starting mixture of RPE cells or progenitors of RPE cells;
• and
• b. separating the cells that are positive for said cell surface marker from cells that are negative for said cell surface marker to obtain a final mixture of RPE cells or progenitors of RPE cells that is more enriched in RPE cells or progenitors of RPE cells compared to the starting mixture;
• wherein said cell surface marker is CD140b, CD56, CD104, CD164, CD220, CD165, EGFR, GD2, CD184, CD10, CD30, CD49a, CD49b, CD50, CD171, TRA-1-60, or CD326.

A method of obtaining Retinal Pigment Epithelium (RPE) cells, comprising:

• culturing one or more human pluripotent stem cells on a first substrate comprising a single first laminin, wherein the laminin is an intact protein or protein fragment;
• exposing the stem cells to a first cell culture medium for a first time period, wherein the first cell culture medium maintains pluripotency;
• removing the first cell culture medium and exposing the stem cells to a second cell culture medium for a second time period of 3 days to 12 days, wherein the second cell culture medium is a differentiation medium and does not contain growth factors;
• removing the second cell culture medium and exposing the stem cells to a third cell culture medium that contains a growth factor being Activin A, for a third time period of 18 days to 35 days to obtain induced RPE cells;
• replating the induced RPE cells on a second substrate comprising a single second laminin;
• and
• culturing the induced RPE cells on the second substrate for 14 days to 28 days to obtain the RPE cells, wherein the total time for differentiation ranges from 42 days to 70 days, and
• wherein the single first laminin and the single second laminin are independently laminin-521, laminin-511, laminin-121, or laminin-111.