Human iPSC-Derived Retinas Recapitulate the Fetal CRB1 CRB2 Complex Formation and Demonstrate that Photoreceptors and Müller Glia Are Targets of AAV5

Stem Cell Reports. 2019 May 14;12(5):906-919. doi: 10.1016/j.stemcr.2019.03.002. Epub 2019 Apr 4.


Human retinal organoids from induced pluripotent stem cells (hiPSCs) can be used to confirm the localization of proteins in retinal cell types and to test transduction and expression patterns of gene therapy vectors. Here, we compared the onset of CRB protein expression in human fetal retina with human iPSC-derived retinal organoids. We show that CRB2 protein precedes the expression of CRB1 in the developing human retina. Our data suggest the presence of CRB1 and CRB2 in human photoreceptors and Müller glial cells. Thus the fetal CRB complex formation is replicated in hiPSC-derived retina. CRB1 patient iPSC retinal organoids showed disruptions at the outer limiting membrane as found in Crb1 mutant mice. Furthermore, AAV serotype 5 (AAV5) is potent in infecting human Müller glial cells and photoreceptors in hiPSC-derived retinas and retinal explants. Our data suggest that human photoreceptors can be efficiently transduced by AAVs in the presence of photoreceptor segments.

Keywords: CRB1; CRB1 patients; CRB2; Müller glial cells; adeno-associated virus; cell polarity; human induced pluripotent stem cells; organoids; photoreceptors; retina.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Cells, Cultured
  • Dependovirus / genetics
  • Ependymoglial Cells / cytology
  • Ependymoglial Cells / metabolism*
  • Ependymoglial Cells / ultrastructure
  • Eye Proteins / genetics
  • Eye Proteins / metabolism*
  • Female
  • Fetus
  • Humans
  • Immunohistochemistry
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Microscopy, Immunoelectron
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Organoids / cytology
  • Organoids / metabolism*
  • Photoreceptor Cells, Vertebrate / metabolism*
  • Photoreceptor Cells, Vertebrate / ultrastructure
  • Pregnancy
  • Retina / cytology
  • Retina / embryology
  • Retina / metabolism*


  • CRB1 protein, human
  • CRB2 protein, human
  • Carrier Proteins
  • Eye Proteins
  • Membrane Proteins
  • Multiprotein Complexes
  • Nerve Tissue Proteins