Skip to content
Back to browse
RETINA Publication-derived

Retinal Organoids for Disease Modelling and Long-Term Culture

Full SOP with Academic or a plan

Source Wahlin et al., 2017 · Wilmer Eye Institute, Johns Hopkins University · 10.1038/s41598-017-00774-9

Authors: Karl J. Wahlin, Julien A. Maruotti, Srinivasa R. Sripathi, John Ball, Juan M. Angueyra, Catherine Kim, Rhonda Grebe, Wei Li, Bryan W. Jones, Donald J. Zack Duration: 300 days 3 phases Cell source: Human iPSC (IMR90.4, EP1), Human ESC (H7)

Abstract

This protocol describes an in vitro method to differentiate human pluripotent stem cells into 3D retinal structures with photoreceptor outer segment-like features, synaptic ribbons, and neurotransmitter expression. The advanced maturation supports modeling of retinal degenerative diseases and screening of neuroprotective compounds.

Cell source
Human iPSC (IMR90.4, EP1), Human ESC (H7)
Application
Disease modeling and developmental study of retinal photoreceptors

Protocol overview

14 steps across 3 phases

Phase 1 · Stem Cell Maintenance and Preparation D-10 to D0
  1. a Culture pluripotent stem cells
  2. b Passage cells with blebbistatin
Phase 2 · Serum-Free Embryoid Body Formation and Optic Vesicle Induction (D0-D18) D0 to D18
  1. a Seed cells for aggregate formation
  2. b Maintain in hypoxia for first day
  3. c Transition to normoxia and begin neural induction
  4. d Optimize medium exchanges and add pathway inhibitors
  5. e Transfer to suspension and enhance retinal induction
  6. f Mechanically isolate optic vesicles
Phase 3 · Long-Term Retinal Cup Development and Maturation (D18-D300) D18 to D300
  1. a Culture isolated optic vesicles in suspension
  2. b Perform periodic grooming and maintenance
  3. c Apply all-trans retinoic acid (ATRA) treatment
  4. d Add Gamma-secretase inhibitor (DAPT)
  5. e Monitor photoreceptor outer segment development
  6. f Continue culture to final maturation stage

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

This SOP was written by Organthis based on the published method in Wahlin et al., 2017. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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