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BRAIN Publication-derived

Retinal Organoids for Developmental Studies (2019)

Full SOP with Academic or a plan

Source Capowski et al. · University of Wisconsin-Madison, Madison, WI, USA · 10.1242/dev.171686

Authors: Capowski EE, Samimi K, Mayber SJ, McUsic AC, Shui YB, Baber MJ, Berlinicke CA, Phillips M, Zack DJ, Gamm DM Duration: 175 days 4 phases Cell source: Human iPSC / Human ESC

Abstract

Capowski et al. established a reproducible, standardized protocol for generating retinal organoids from human pluripotent stem cells with defined morphological staging criteria enabling rigorous quality control. The Gamm lab Wisconsin protocol validated differentiation across multiple cell lines and provides the reference framework for retinal organoid research and clinical-grade manufacturing.

Cell source
Human iPSC / Human ESC
Application
Standardized Retinal Organoid Production

Protocol overview

20 steps across 4 phases

Phase 1 · hPSC maintenance and embryoid body formation Day 0 - Day 7
  1. a Maintain pluripotent stem cells
  2. b Lift embryoid bodies
  3. c Wean EBs into Neural Induction Medium
  4. d Add BMP4 to culture
  5. e Plate EBs on Matrigel
  6. f Perform sequential half media changes
Phase 2 · Optic vesicle formation and transition to retinal differentiation Day 16 - Day 30
  1. a Switch to Retinal Differentiation Medium
  2. b Monitor optic vesicle formation
  3. c Dissect optic vesicles
Phase 3 · Suspension culture and long-term organoid maturation Day 30 - Day 175 or longer
  1. a Transfer organoids to poly-HEMA-coated flasks
  2. b Feed organoids with 3D-RDM twice weekly
  3. c Add all-trans retinoic acid
  4. d Remove retinoic acid
  5. e Monitor organoid morphology by light microscopy
  6. f Culture organoids for extended maturation
Phase 4 · Organoid characterization and analysis Day 35 onwards
  1. a Perform optical coherence tomography (OCT)
  2. b Perform immunocytochemistry
  3. c Perform electron microscopy (TEM)
  4. d Perform multi-photon imaging
  5. e Perform optical metabolic imaging (OMI)

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Attribution

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

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