Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large Animal Model

Laurence  M. Occelli; Felipe Marinho; Ratnesh  K. Singh; Francois Binette; Igor  O. Nasonkin; Simon M. Petersen-Jones

doi:10.3791/61683

Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large ...

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JoVE Journal Neuroscience

Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large Animal Model

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07:43 min

August 5, 2021

DOI: 10.3791/61683-v

Laurence M. Occelli*¹, Felipe Marinho*¹, Ratnesh K. Singh², Francois Binette², Igor O. Nasonkin², Simon M. Petersen-Jones¹

¹College of Veterinary Medicine, Department of Small Animal Clinical Sciences,Michigan State University, ²Lineage Cell Therapeutics, Inc.

Overview

This article presents a surgical technique for transplanting human pluripotent stem cell (hPSC)-derived retinal tissue into the subretinal space of a large animal model. The method focuses on direct delivery of retinal organoids to target tissues, with the goal of developing cell therapy for retinal degenerative diseases.

Key Study Components

Area of Science

Neuroscience
Stem Cell Therapy
Retinal Degeneration

Background

Retinal diseases cause significant vision loss worldwide.
Cell therapy using hPSC-derived tissues holds potential for treatment.
Techniques for precise tissue delivery are critical for efficacy.
This study outlines a method to improve implantation success rates.

Purpose of Study

To develop a reliable method for subretinal delivery of retinal organoids.
To assess the feasibility of cell therapy approaches for retinal degeneration.
To demonstrate the procedure using a large animal model.

Methods Used

A surgical technique is performed on a four-month-old anesthetized cat.
Direct implantation of hPSC-derived retinal organoids into the subretinal space.
Key steps include lateral canthotomy, vitrectomy, and careful handling of ocular tissues.
Post-operative examinations confirm organoid placement and viability.

Main Results

The technique successfully delivers retinal organoids to the targeted retinal area.
Surgeon visualization confirms correct placement during and after the procedure.
Results suggest potential for further exploration of stem cell treatments in clinical settings.

Conclusions

This study establishes a surgical protocol for retinal organoid transplantation.
The approach provides a foundation for future research in cell therapy for retinal diseases.
Successful implantation techniques could enhance our understanding of retinal repair mechanisms.

Frequently Asked Questions

What advantages does this method offer for retinal therapy?

This surgical technique allows for direct delivery of retinal organoids to the target site, potentially increasing the efficacy of cell therapy in treating retinal degenerative conditions.

How is the retinal organoid transplantation performed?

The process involves a detailed surgical protocol including canthotomy, conjunctival suturing, sclerotomy, and organoid injection through an adequately sized vitrectomy port.

What outcomes can be measured after the procedure?

Outcomes include successful placement of organoids within the subretinal space, confirmed via post-operative ophthalmic exams and imaging techniques.

What is the significance of using a large animal model?

Using a large animal model like a cat allows for better translation of surgical techniques to human applications due to anatomical similarities.

Are there any limitations to this approach?

Limitations may include the need for precise surgical skills and the risk of complications during the procedure, necessitating careful planning and execution.

Presented here is a surgical technique for transplanting human pluripotent stem cell (hPSC)-derived retinal tissue into the subretinal space of a large animal model.

The current protocol shows in detail subretinal delivery of the whole retinal organoids in a large animal model, aiming at developing cell therapy approaches for retinal degeneration. The main advantage of this technique is that it allows for delivery of organoids directly to the target tissue, the retina. The technique aims to treat advanced blinding retinal degenerative conditions, such as age-related macular degeneration, retinitis pigmentosa, or leber congenital amaurosis.

Demonstrating the procedure will be Dr.Peterson Jones. For subretinal implantation of the organoids, use Stevens Tenotomy Scissors to perform a 0.5 to 1 centimeter lateral canthotomy in a four-month-old overnight-fasted anesthetized cat, and place an appropriately sized Barraquer eyelid speculum into the incision to keep the eyelids open with a surgical assistant regularly irrigating the cornea with BSS throughout the procedure. Use 0.5 Castroviejo corneal tying forceps and a small mosquito hemostat to gently grasp the bulbar conjunctiva next to the limbus.

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