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Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells
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Biologie du développement
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Journal JoVE Biologie du développement
Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

DOI:
10.3791/64509-v

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09:47 min

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December 09, 2022

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, , , , ,
1Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre,LV Prasad Eye Institute, 2Manipal Academy of Higher Education, 3Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School

Chapitres

  • 00:05Introduction
  • 01:19Passaging and Plating of hiPSCs to Initiate Retinal Lineage Differentiation
  • 02:40Differentiation of hiPSCs into Eye Fields and Retinal Lineage
  • 04:38Harvesting of Retinal Organoids
  • 06:41Results: Characterization of Retinal Organoids and RPE Cells Derived From Normal and RB1-/- Mutant iPSCs
  • 09:06Conclusion

Summary

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This protocol describes an efficient method of differentiating hiPSCs into eye field clusters and generating neuro-retinal organoids using simplified culture conditions involving both adherent and suspension culture systems. Other ocular cell types, such as the RPE and corneal epithelium, can also be isolated from mature eye fields in retinal cultures.

Tags

Keywords: Retinal OrganoidsHuman Induced Pluripotent Stem CellsRetinal DegenerationRetinal Pigment EpitheliumIn Vitro ModelsCell-based TherapiesEye DevelopmentInherited Retinal DiseasesDifferentiationAdherent CultureSuspension CultureRetinal CupsSpatial PositioningMorphological FeaturesCell DissociationIPSC CultureDifferentiation Induction MediumBFGFNoggin

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