Imaging Transplanted Photoreceptors in Living Nonhuman Primates with Single-Cell Resolution

Ebrahim Aboualizadeh; M Joseph Phillips; Juliette E McGregor; David A DiLoreto Jr; Jennifer M Strazzeri; Kamal R Dhakal; Brittany Bateman; Lindsey D Jager; Kelsy L Nilles; Sara A Stuedemann; Allison L Ludwig; Jennifer J Hunter; William H Merigan; David M Gamm; David R Williams

doi:10.1016/j.stemcr.2020.06.019

Imaging Transplanted Photoreceptors in Living Nonhuman Primates with Single-Cell Resolution

Stem Cell Reports. 2020 Aug 11;15(2):482-497. doi: 10.1016/j.stemcr.2020.06.019. Epub 2020 Jul 23.

Authors

Affiliations

¹ Center for Visual Science, University of Rochester, Rochester, NY, USA.
² Waisman Center, University of Wisconsin, Madison, WI, USA; McPherson Eye Research Institute, University of Wisconsin, Madison, WI, USA.
³ Center for Visual Science, University of Rochester, Rochester, NY, USA; Flaum Eye Institute, University of Rochester, Rochester, NY, USA.
⁴ Flaum Eye Institute, University of Rochester, Rochester, NY, USA.
⁵ Waisman Center, University of Wisconsin, Madison, WI, USA.
⁶ Center for Visual Science, University of Rochester, Rochester, NY, USA; Flaum Eye Institute, University of Rochester, Rochester, NY, USA; The Institute of Optics, University of Rochester, Rochester, NY, USA.
⁷ Waisman Center, University of Wisconsin, Madison, WI, USA; McPherson Eye Research Institute, University of Wisconsin, Madison, WI, USA; Department of Ophthalmology and Visual Sciences, University of Wisconsin, Madison, WI, USA.
⁸ Center for Visual Science, University of Rochester, Rochester, NY, USA; The Institute of Optics, University of Rochester, Rochester, NY, USA. Electronic address: david.williams@rochester.edu.

Abstract

Stem cell-based transplantation therapies offer hope for currently untreatable retinal degenerations; however, preclinical progress has been largely confined to rodent models. Here, we describe an experimental platform for accelerating photoreceptor replacement therapy in the nonhuman primate, which has a visual system much more similar to the human. We deployed fluorescence adaptive optics scanning light ophthalmoscopy (FAOSLO) to noninvasively track transplanted photoreceptor precursors over time at cellular resolution in the living macaque. Fluorescently labeled photoreceptors generated from a CRX^+/tdTomato human embryonic stem cell (hESC) reporter line were delivered subretinally to macaques with normal retinas and following selective ablation of host photoreceptors using an ultrafast laser. The fluorescent reporter together with FAOSLO allowed transplanted photoreceptor precursor survival, migration, and neurite formation to be monitored over time in vivo. Histological examination suggested migration of photoreceptor precursors to the outer plexiform layer and potential synapse formation in ablated areas in the macaque eye.

Keywords: adaptive optics retinal imaging; fluorescence; hPSC; integration and survival; in vivo; nonhuman primates; photoreceptor precursor; retinal degeneration; stem cell therapy; ultrafast.

Publication types

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

MeSH terms

Animals
Cell Differentiation
Fluorescence
Humans
Light
Models, Animal
Optics and Photonics
Photoreceptor Cells / transplantation*
Primates
Retina / metabolism
Single-Cell Analysis
Tomography, Optical Coherence

Abstract

Publication types

MeSH terms

Grants and funding