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RETINA Publication-derived

Retinal Organoids for Disease and Development (2021)

Full SOP with Academic or a plan

Source Afanasyeva et al., 2021 · Radboud University Medical Center · 10.1007/s00018-021-03917-4

Authors: Tess A. V. Afanasyeva, Julio C. Corral-Serrano, Alejandro Garanto, Ronald Roepman, Michael E. Cheetham, Rob W. J. Collin Duration: 180 days 4 phases Cell source: Human iPSC

Abstract

This protocol generates three-dimensional retinal organoids from human induced pluripotent stem cells (iPSCs) that contain all major retinal cell types including photoreceptors, ganglion cells, bipolar cells, horizontal cells, amacrine cells, and Müller glia. The organoids recapitulate many aspects of human retinal development and can be used to model inherited retinal diseases, test gene therapies including AAV-based approaches and antisense oligonucleotides, and assess photoreceptor transplantation strategies.

Cell source
Human iPSC
Application
Disease modeling and therapeutic development for inherited retinal diseases

Protocol overview

12 steps across 4 phases

Phase 1 · Embryoid Body Formation and Neural Induction D0-D16
  1. a Initiate embryoid body formation
  2. b Apply neural induction medium at D1
  3. c Continue neural induction through D16
Phase 2 · Retinal Differentiation and Early Organoid Development D16-D63
  1. a Transition to retinal differentiation medium
  2. b Add supplementary factors for photoreceptor development (D25-D63)
  3. c Maintain organoid culture in suspension
Phase 3 · Photoreceptor Maturation and Late Differentiation D63-D180+
  1. a Begin retinoic acid supplementation
  2. b Assess organoid maturation stages
  3. c Continue culture through maturation period
  4. d Optional: Add N2 supplement in later stages
Phase 4 · Optional: AAV Transduction for Gene Therapy Studies D44-D180
  1. a Prepare AAV vectors for transduction
  2. b Add AAV to organoid culture

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Attribution

This SOP was written by Organthis based on the published method in Afanasyeva et al., 2021. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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