title: Retinal Organoids from Pluripotent Stem Cells Efficiently Recapitulate Retinogenesis.
abstract: The plasticity of pluripotent stem cells provides new possibilities for studying development, degeneration, and regeneration. Protocols for the differentiation of retinal organoids from embryonic stem cells have been developed, which either recapitulate complete eyecup morphogenesis or maximize photoreceptor genesis. Here, we have developed a protocol for the efficient generation of large, 3D-stratified retinal organoids that does not require evagination of optic-vesicle-like structures, which so far limited the organoid yield. Analysis of gene expression in individual organoids, cell birthdating, and interorganoid variation indicate efficient, reproducible, and temporally regulated retinogenesis. Comparative analysis of a transgenic reporter for PAX6, a master regulator of retinogenesis, shows expression in similar cell types in mouse in vivo, and in mouse and human retinal organoids. Early or late Notch signaling inhibition forces cell differentiation, generating organoids enriched with cone or rod photoreceptors, respectively, demonstrating the power of our improved organoid system for future research in stem cell biology and regenerative medicine.
authors.affiliation: German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307 Dresden, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Völkner
authors.affiliation: German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307 Dresden, Germany.
authors.forename: Marlen
authors.initials: M
authors.lastname: Zschätzsch
authors.affiliation: Stem Cell Engineering, Biotechnology Center (BIOTEC), TU Dresden, 01307 Dresden, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Rostovskaya
authors.affiliation: CRTD - Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307 Dresden, Germany.
authors.forename: Rupert W
authors.initials: RW
authors.lastname: Overall
authors.affiliation: CRTD - Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307 Dresden, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Busskamp
authors.affiliation: Stem Cell Engineering, Biotechnology Center (BIOTEC), TU Dresden, 01307 Dresden, Germany.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Anastassiadis
authors.affiliation: German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307 Dresden, Germany; CRTD - Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307 Dresden, Germany. Electronic address: mike.karl@dzne.de.
authors.forename: Mike O
authors.initials: MO
authors.lastname: Karl
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D009924:Organ Culture Techniques; D038081:Organogenesis / Q000235:genetics; D009940:Organoids / Q000166:cytology / Q000378:metabolism; D000071841:PAX6 Transcription Factor / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012160:Retina / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000071841:PAX6 Transcription Factor