Generation of three-dimensional retinal organoids expressing rhodopsin and S- and M-cone opsins from mouse stem cells

Biochem Biophys Res Commun. 2018 Jan 22;495(4):2595-2601. doi: 10.1016/j.bbrc.2017.12.092. Epub 2017 Dec 20.


Purpose: Three-dimensional retinal organoids can be differentiated from embryonic stem cells/induced pluripotent stem cells (ES/iPS cells) under defined medium conditions. We modified the serum-free floating culture of embryoid body-like aggregates with quick reaggregation (SFEBq) culture procedure to obtain retinal organoids expressing more rod photoreceptors and S- and M-cone opsins.

Methods: Retinal organoids differentiated from mouse Nrl-eGFP iPS cells were cultured in various mediums during photoreceptor development. To promote rod photoreceptor development, organoids were maintained in media containing 9-cis retinoic acids (9cRA). To obtain retinal organoids with M-opsin expression, we cultured in medium with 1% fetal bovine serum (FBS) supplemented with T3, BMP4, and DAPT. Section immunohistochemistry was performed to visualize the expression of photoreceptor markers.

Results: In three-dimensional (3D) retinas exposed to 9cRA, rhodopsin was expressed earlier and S-cone opsins were suppressed. We could maintain 3D retinas up to DD 35 in culture media with 1% FBS. The 3D retinas expressed rhodopsin, S- and M-opsins, but most cone photoreceptors expressed either S- or M-opsins.

Conclusion: By modifying culture conditions in the SFEBq protocol, we obtained rod-dominated 3D retinas and S- and M-opsin expressing 3D retinas.

Keywords: 3D retinal aggregates; Cone photoreceptor; Retinoic acid; Rod photoreceptor; Stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bioartificial Organs*
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Cone Opsins / metabolism*
  • Mice
  • Organogenesis / physiology
  • Printing, Three-Dimensional
  • Retina / cytology*
  • Retina / growth & development*
  • Retinal Cone Photoreceptor Cells / metabolism
  • Rhodopsin / metabolism*
  • Stem Cells / cytology*
  • Stem Cells / physiology
  • Tissue Engineering / methods*


  • Cone Opsins
  • Rhodopsin