Micromanipulator-Assisted Subretinal Transplantation of Human Photoreceptor Reporter Cell Suspensions into Mice

Research output: Contribution to book/conference proceedings/anthology/reportChapter in book/anthology/reportContributedpeer-review

Contributors

Abstract

Retinopathies marked by photoreceptor cell loss are the main cause of visual impairment and blindness in industrialized societies. Photoreceptor replacement strategies are examined as potential treatment options, for instance, by subretinal transplantation of photoreceptor-enriched cell suspensions, an approach which has shown beneficial effects in mouse models of retinal degeneration. As it is now possible to generate human photoreceptors through retinal organoid culture, preclinical research has shifted to human-into-mouse transplantations. Here, we describe a detailed method for human retinal organoid dissociation and fluorescent reporter-based sorting of photoreceptors. We introduce a transvitreal, micromanipulator-based transplantation technique to increase the quality and efficiency of cell injections into the mouse subretinal space, thereby increasing reproducibility and reducing experimental animal numbers. Lastly, we use a local corticosteroid to prevent immune rejection of the grafted cells in this human-into-mouse xenotransplantation setting.

Details

Original languageEnglish
Title of host publicationNeuromethods
PublisherHumana Press
Pages81-98
Number of pages18
Publication statusPublished - 2023
Peer-reviewedYes

Publication series

SeriesNeuromethods
Volume189
ISSN0893-2336

External IDs

ORCID /0000-0001-9467-7677/work/161888202

Keywords

Sustainable Development Goals

Keywords

  • 3R, Flow cytometry, Micromanipulator, Photoreceptor replacement, Reduction, Refinement, Reporter iPSC, Retinal organoid, Subretinal, Suspension, Transplantation