Successful applications of pluripotent stem cells to cell-based therapies will rely on rapid and efficient methods to differentiate cells toward the target cell type. While methods have been developed for the generation of some medically relevant cell types including retinal pigment epithelium (RPE) cells, such protocols are lengthy and result in a heterogeneous cell mixture of RPE and non-RPE cells, requiring manual subselection and expansion. Such considerations have significant limiting impact of therapeutic applicability. Here we describe the accelerated three-dimensional neuroepithelial cyst culture of human embryonic stem cells (hESCs) and its utility to achieve quantitative production of RPE cell sheet with no manual selection in 30 days.