Generation of Three-Dimensional Retinal Organoids from Human Pluripotent Stem Cells
Source Afanasyeva et al., 2021 · Radboud University Medical Center · 10.1007/s00018-021-03917-4
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.
Protocol overview
12 steps across 4 phases
- 1 Initiate embryoid body formation
- 2 Apply neural induction medium at D1
- 3 Continue neural induction through D16
- 4 Transition to retinal differentiation medium
- 5 Add supplementary factors for photoreceptor development (D25-D63)
- 6 Maintain organoid culture in suspension
- 7 Begin retinoic acid supplementation
- 8 Assess organoid maturation stages
- 9 Continue culture through maturation period
- 10 Optional: Add N2 supplement in later stages
- 11 Prepare AAV vectors for transduction
- 12 Add AAV to organoid culture
Full SOP
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Attribution
This SOP was authored by Organthis based on the published method in Afanasyeva et al., 2021. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.
This wording is awaiting legal review.
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