Three-dimensional early retinal progenitor 3D tissue constructs derived from human embryonic stem cells

J Neurosci Methods. 2010 Jun 30;190(1):63-70. doi: 10.1016/j.jneumeth.2010.04.025. Epub 2010 May 4.


Purpose: To develop three-dimensional (3D) constructs of retinal pigment epithelium (RPE) and early retina progenitor cells from human embryonic stem cells (hESCs).

Methods: 3D tissue constructs were developed by culturing hESC-derived neural retinal progenitors in a matrix on top of hESC-derived RPE cells in a cell culture insert. An osmolarity gradient maintained the nutrition of the 3D cell constructs. Cross-sections through hESC-derived tissue constructs were characterized by immunohistochemistry for various transcription factors and cell markers.

Results: hESC-derived tissue constructs expressed transcription factors characteristic of retinal development, such as pax6, Otx2, Chx10, retinal RAX; Brn3b (necessary for differentiation of retinal ganglion cells); and crx and nrl (role in photoreceptor development). Many cells expressed neuronal markers including nestin, beta-tubulin and microtubule-associated proteins.

Conclusions: This study shows for the first time that 3D early retinal progenitor tissue constructs can be derived from hESCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Line
  • Cell Proliferation
  • Embryonic Stem Cells / physiology*
  • Extracellular Matrix
  • Humans
  • Immunohistochemistry
  • Mice
  • Neurons / physiology
  • Retina / physiology*
  • Retinal Pigment Epithelium / physiology*
  • Tissue Engineering / methods*
  • Tissue Scaffolds
  • Transcription Factors / metabolism


  • Transcription Factors