Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

J Vis Exp. 2017 Oct 30:(128):56274. doi: 10.3791/56274.

Abstract

We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing a detailed and thorough protocol is to clearly demonstrate each step and to make this readily available to researchers in the field. This protocol results in a homogenous layer of RPE with minimal or no manual dissection needed. The method presented here has been shown to be effective for induced pluripotent stem cells (iPSC) and human embryonic stem cells. Additionally, we describe methods for cryopreservation of intermediate cell banks that allow long-term storage. RPE generated using this protocol might be useful for iPSC disease-in-a-dish modeling or clinical application.

Publication types

  • Video-Audio Media

MeSH terms

  • Cell Differentiation / physiology
  • Cytological Techniques / methods*
  • Embryonic Stem Cells
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Retinal Pigment Epithelium / cytology*