Skip to content
← Back to browse
RETINA Publication-derived

Human iPSC differentiation to retinal organoids in response to IGF1 and BMP4 activation

Source Chichagova et al., 2020 · Institute of Genetic Medicine, Newcastle University · 10.1002/stem.3116

👤 Valeria Chichagova, Gerrit Hilgen, Ali Ghareeb, Maria Georgiou, Madeleine Carter, Evelyne Sernagor, Majlinda Lako, Lyle Armstrong ⏱ 180 days 📋 7 phases 🧫 Human iPSC

Abstract

This protocol describes the generation of retinal organoids from human iPSC lines using two distinct differentiation methods that modulate either BMP4 (Method I) or IGF1 (Method II) signaling pathways. The protocol produces three-dimensional retinal structures containing photoreceptors, retinal ganglion cells, Müller glia, and other retinal cell types with variable efficiency depending on iPSC line and method chosen. The organoids demonstrate light-responsive functional properties and serve as platforms for disease modeling and developmental studies.

Cell source
Human iPSC
Application
Disease modeling and developmental study of retinal organogenesis

Protocol overview

13 steps across 7 phases

iPSC Maintenance and Pluripotency Confirmation Prior to differentiation
  1. 1 Expand iPSC lines to required cell density
Method I: BMP4 Pathway Activation Days 0–18 (initial phase)
  1. 1 Initiate BMP4-dependent retinal differentiation
Method II: IGF1 Pathway Activation Days 0–18 (initial phase)
  1. 1 Initiate IGF1-dependent retinal differentiation
Maturation Phase with Supplementary Factors (Day 18 onwards) Days 18–180+
  1. 1 Add supplementary maturation factors
  2. 2 Monitor morphological development through day 85 and day 169
  3. 3 Continue culture to day 180 for gene and protein expression analysis
Gene Expression Analysis (Day 180) Day 180
  1. 1 Extract RNA and perform quantitative PCR
Protein Expression and Localization Analysis (Day 180) Day 180
  1. 1 Perform immunofluorescence microscopy for photoreceptor markers
  2. 2 Stain for non-photoreceptor retinal cell types
Functional Assessment: Light-Driven Responses (Days 150–180+) Days 150–180+
  1. 1 Prepare organoids for electrophysiological recording
  2. 2 Apply white light stimulus and record RGC responses
  3. 3 Apply sustained blue light stimulus to identify intrinsically photosensitive RGCs
  4. 4 Quantify change of firing (COF) and compare between methods

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Chichagova et al., 2020. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence