Degenerative disorders of the retina (including age-related macular degeneration), which originate primarily at or within the retinal pigmented epithelial (RPE) layer, lead to a progressive disorganization of the retinal anatomy and the deterioration of visual function. The substitution of damaged RPE cells (RPEs) with in vitro cultured RPE cells using a subretinal cell carrier has shown potential for re-establishing the anatomical structure of the outer retinal layers and is, therefore, being further studied. Here, we present the principles of a surgical technique that allows for the effective subretinal transplantation of a cell carrier with cultivated RPEs into minipigs. The surgeries were performed under general anesthesia and included a standard lens-sparing three-port pars plana vitrectomy (PPV), subretinal application of a balanced salt solution (BSS), a 2.7 mm retinotomy, implantation of a nanofibrous cell carrier into the subretinal space through an additional 3.0 mm sclerotomy, fluid-air exchange (FAX), silicone oil tamponade, and closure of all the sclerotomies. This surgical approach was used in 29 surgeries (18 animals) over the past 8 years with a success rate of 93.1%. Anatomic verification of the surgical placement was carried out using in vivo fundus imaging (fundus photography and optical coherence tomography). The recommended surgical steps for the subretinal implantation of RPEs on a carrier in minipig eyes can be used in future preclinical studies using large-eye animal models.

Center for Eye Research Department of Ophthalmology Oslo University Hospital and Institute of Clinical Medicine Faculty of Medicine University of Oslo

Department of Ophthalmology University Hospital Kralovske Vinohrady and 3rd Faculty of Medicine Charles University Prague

Eye Center Donaustadt Department of Ophthalmology Sigmund Freud University

Eye Clinic Department of Ophthalmology University Hospital Giessen and Marburg GmbH

Eye Clinic Department of Ophthalmology University Hospital Giessen and Marburg GmbH; Karl Landsteiner Institute for Retinal Research and Imaging;

Institute of Animal Physiology and Genetics Czech Academy of Sciences

Institute of Animal Physiology and Genetics Czech Academy of Sciences; Department of Cell Biology Faculty of Science Charles University

Institute of Experimental Medicine Czech Academy of Sciences

Institute of Macromolecular Chemistry Czech Academy of Sciences

Moorfields Eye Hospitals UAE

Stem Cell Therapies in Neurodegenerative Diseases Lab Research Center Principe Felipe

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