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Device-free isolation of photoreceptor cells from patient iPSC-derived retinal organoids
Journal article   Peer reviewed

Device-free isolation of photoreceptor cells from patient iPSC-derived retinal organoids

Nicholas E Stone, Laura R Bohrer, Nathaniel K Mullin, Alexander Berthold, Allison T Wright, Ian C Han, Edwin M Stone, Robert F Mullins and Budd A Tucker
JCI insight, Vol.10(14), e186338
07/22/2025
DOI: 10.1172/jci.insight.186338
PMCID: PMC12288960
PMID: 40504625

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Abstract

Autologous photoreceptor cell replacement is one of the most promising strategies currently being developed for the treatment of patients with inherited retinal degenerative blindness. Induced pluripotent stem cell (iPSC) derived retinal organoids, which faithfully recapitulate the structure of the neural retina, are an ideal source of transplantable photoreceptors required for these therapies. However, retinal organoids contain other retinal cell types, including bipolar, horizontal and amacrine cells, which are unneeded and may reduce the potency of the final therapeutic product. Therefore, approaches for isolating fate committed photoreceptor cells from dissociated retinal organoids are desirable. In this work, we present partial dissociation, a technique which leverages the high level of organization found in retinal organoids to enable selective enrichment of photoreceptor cells without the use of specialized equipment or reagents such as antibody labels. We demonstrate up to 90% photoreceptor cell purity by simply selecting cell fractions liberated from retinal organoids during enzymatic digestion in the absence of mechanical dissociation. As the presented approach relies on the use of standard plasticware and commercially available cGMP compliant reagents, we believe that it is ideal for use in the preparation of clinical photoreceptor cell replacement therapies.
Translation Ophthalmology Stem Cells iPS cells

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