July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Characterization of interphotoreceptor matrix proteins expressed by human pluripotent stem cell derived retinal cells
Author Affiliations & Notes
  • Steven Mayerl
    Cellular and Molecular Pathology, University of Wisconsin-Madison, Madison, Wisconsin, United States
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Elizabeth E. Capowski
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Divya Sinha
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Joe Phillips
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Kimberly L Edwards
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • David M Gamm
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Steven Mayerl, None; Elizabeth Capowski, None; Divya Sinha, None; Joe Phillips, None; Kimberly Edwards, None; David Gamm, None
  • Footnotes
    Support  Foundation Fighting Blindness, Research to Prevent Blindness, Emmett A. Humble Distinguished Directorship of the Retina Research Foundation, McPherson Eye Research Institute Sandra Lemke Trout Chair in Eye Research, NICHD Grant U54 HD090256
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 3339. doi:
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    • Get Citation

      Steven Mayerl, Elizabeth E. Capowski, Divya Sinha, Joe Phillips, Kimberly L Edwards, David M Gamm; Characterization of interphotoreceptor matrix proteins expressed by human pluripotent stem cell derived retinal cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3339.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Human pluripotent stem cells (hPSCs) are powerful tools for modeling retinal development and disease. Interphotoreceptor matrix (IPM) proteins play an important role in the maturation of photoreceptors and, when defective, can lead to photoreceptor degenerative diseases. The objective of this study was to assess the expression and localization of IPM proteins in hPSC-derived retinal cells.

Methods : Three hPSC lines were differentiated to retinal organoids and maintained in culture until a developmental time point where mature photoreceptors were observed (stage 3; 150-250 days). Retinal organoids were then fixed, cryosectioned at 15μm thickness, and immunostained using primary antibodies against IPM and photoreceptor-specific proteins. Imaging was performed using a Nikon A1R-SI confocal microscope. Immunostained cryosections of non-human primate retina were used for comparison.

Results : Retinal organoids derived from hPSC-derived retinal organoids express and properly localize insoluble IPM markers to their outer surface where photoreceptor outer segments abound, similar to what is observed in non-human primate retina. IPM proteins detected in these stage 3 retinal organoids include PNA, WGA, IMPG1, IMPG2, RP1, IRBP, Versican, and Brevican. However, differences between retinal organoids and non-human primate retina were found with regard to localization of soluble IPM proteins, which is most likely due to the lack of a sequestered subretinal space in free-floating organoid cultures.

Conclusions : We demonstrate that hPSC-derived retinal organoids express and appropriately localize insoluble IPM proteins along their outer surface. Characterization of the IPM in hPSC-derived retinal organoids provides a platform to model photoreceptor maturation and some forms of inherited retinal degenerative disorders.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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