Role of RB1 in human embryonic stem cell-derived retinal organoids

Dev Biol. 2020 Jun 15;462(2):197-207. doi: 10.1016/j.ydbio.2020.03.011. Epub 2020 Mar 19.


Three-dimensional (3D) organoid models derived from human pluripotent stem cells provide a platform for studying human development and understanding disease mechanisms. Most studies that examine biallelic inactivation of the cell cycle regulator Retinoblastoma 1 (RB1) and the link to retinoblastoma is in mice, however, less is known regarding the pathophysiological role of RB1 during human retinal development. To study the role of RB1 in early human retinal development and tumor formation, we generated retinal organoids from CRISPR/Cas9-derived RB1-null human embryonic stem cells (hESCs). We showed that RB is abundantly expressed in retinal progenitor cells in retinal organoids and loss of RB1 promotes S-phase entry. Furthermore, loss of RB1 resulted in widespread apoptosis and reduced the number of photoreceptor, ganglion, and bipolar cells. Interestingly, RB1 mutation in retinal organoids did not result in retinoblastoma formation in vitro or in the vitreous body of NOD/SCID immunodeficient mice. Together, our work identifies a crucial function for RB1 in human retinal development and suggests that RB1 deletion alone is not sufficient for tumor development, at least in human retinal organoids.

Keywords: CRISPR/Cas9; Cell cycle; Human embryonic stem cells; Organoid; Retinal disease; Retinoblastoma.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / physiology
  • CRISPR-Cas Systems
  • Cell Differentiation / genetics
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Organoids / cytology
  • Pluripotent Stem Cells / cytology
  • Retina / embryology*
  • Retina / physiology
  • Retinal Ganglion Cells / metabolism
  • Retinal Neoplasms / metabolism
  • Retinoblastoma / metabolism
  • Retinoblastoma Binding Proteins / metabolism*
  • Retinoblastoma Binding Proteins / physiology
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitin-Protein Ligases / physiology


  • RB1 protein, human
  • Retinoblastoma Binding Proteins
  • Ubiquitin-Protein Ligases