Method for producing polarized retinal progenitor cells from pluripotent stem cells and their differentiation into retinal pigment epithelium cells

摘要

The present invention relates to a method and components for producing polarized retinal progenitor cells (RPC) from pluripotent stem cells in high yield and purity. The polarized retinal progenitor cells are preferably further differentiated with high efficiency and speed into retinal pigment epithelium cells (RPE cells). The cells obtained are particularly suitable for use in cell transplantation or in the generation of transplant tissue and are particularly applicable to screening systems for substances that modulate the function of polarized retinal progenitor cells and/or RPE cells.;In the method according to the invention pluripotent stem cells are differentiated into polarized retinal progenitor cells by first culturing pluripotent stem cell colonies in three-dimensional cell culture embedded in a proteinaceous gel comprising at least two proteins selected from the group consisting of Laminin, Collagen IV Entactin and Perlecan and comprising at least two growth factors selected from agonists of the EGF, FGF2, NGF, PDGF, IGF-1 and TGF-beta pathways until polarized neural cysts develop and subsequently dissociating the polarized neural cysts into dispersed polarized retinal progenitor cells.