Generation of Rx+/Pax6+ neural retinal precursors from embryonic stem cells

Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11331-6. doi: 10.1073/pnas.0500010102. Epub 2005 Aug 2.


We report directed differentiaion of retinal precursors in vitro from mouse ES cells. Six3+ rostral brain progenitors are generated by culturing ES cells under serum-free suspension conditions (SFEB culture) in the presence of Wnt and Nodal antagonists (Dkk1 and LeftyA), and subsequently steered to differentiate into Rx+ cells (16%) by treatment with activin and serum. Consistent with the characteristics of early neural retinal precursors, the induced Rx+ cells coexpress Pax6 and the mitotic marker Ki67, but not Nestin. The ES cell-derived precursors efficiently generate cells with the photoreceptor phenotype (rhodopsin+, recoverin+) when cocultured with embryonic retinal cells. Furthermore, organotypic culture studies demonstrate the selective integration and survival of ES cell-derived cells with the photoreceptor phenotype (marker expression and morphology) in the outer nuclear layer of the retina. Taken together, ES cells treated with SFEB/Dkk1/LeftyA/serum/activin generate neural retinal precursors, which have the competence of photoreceptor differentiation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activins / pharmacology
  • Animals
  • Cell Differentiation / drug effects*
  • Cells, Cultured
  • Culture Media, Serum-Free / pharmacology
  • Eye Proteins / metabolism*
  • Genetic Vectors
  • Homeodomain Proteins / metabolism*
  • Immunohistochemistry
  • In Situ Hybridization
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Left-Right Determination Factors
  • Lentivirus
  • Mice
  • Nerve Tissue Proteins / metabolism
  • PAX6 Transcription Factor
  • Paired Box Transcription Factors / metabolism*
  • Repressor Proteins / metabolism*
  • Retina / cytology*
  • Retina / metabolism
  • Rhodopsin / metabolism
  • Serum
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Transforming Growth Factor beta / pharmacology


  • Culture Media, Serum-Free
  • Dkk1 protein, mouse
  • Eye Proteins
  • Homeodomain Proteins
  • Intercellular Signaling Peptides and Proteins
  • Left-Right Determination Factors
  • Nerve Tissue Proteins
  • PAX6 Transcription Factor
  • Paired Box Transcription Factors
  • Pax6 protein, mouse
  • Repressor Proteins
  • Rx protein, mouse
  • Sine oculis homeobox homolog 3 protein
  • Transforming Growth Factor beta
  • Activins
  • Rhodopsin