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Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture

Using retinal organoid systems, organ-like 3D tissues, relies implicitly on their robustness. However, essential key parameters, particularly retinal growth and longer-term culture, are still insufficiently defined. Here, we hypothesize that a previously optimized protocol for high yield of evenly-s...

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Autores principales: Völkner, Manuela, Kurth, Thomas, Schor, Jana, Ebner, Lynn J. A., Bardtke, Lara, Kavak, Cagri, Hackermüller, Jörg, Karl, Mike O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8114082/
https://www.ncbi.nlm.nih.gov/pubmed/33996806
http://dx.doi.org/10.3389/fcell.2021.645704
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author Völkner, Manuela
Kurth, Thomas
Schor, Jana
Ebner, Lynn J. A.
Bardtke, Lara
Kavak, Cagri
Hackermüller, Jörg
Karl, Mike O.
author_facet Völkner, Manuela
Kurth, Thomas
Schor, Jana
Ebner, Lynn J. A.
Bardtke, Lara
Kavak, Cagri
Hackermüller, Jörg
Karl, Mike O.
author_sort Völkner, Manuela
collection PubMed
description Using retinal organoid systems, organ-like 3D tissues, relies implicitly on their robustness. However, essential key parameters, particularly retinal growth and longer-term culture, are still insufficiently defined. Here, we hypothesize that a previously optimized protocol for high yield of evenly-sized mouse retinal organoids with low variability facilitates assessment of such parameters. We demonstrate that these organoids reliably complete retinogenesis, and can be maintained at least up to 60 days in culture. During this time, the organoids continue to mature on a molecular and (ultra)structural level: They develop photoreceptor outer segments and synapses, transiently maintain its cell composition for about 5–10 days after completing retinogenesis, and subsequently develop pathologic changes – mainly of the inner but also outer retina and reactive gliosis. To test whether this organoid system provides experimental access to the retina during and upon completion of development, we defined and stimulated organoid growth by activating sonic hedgehog signaling, which in patients and mice in vivo with a congenital defect leads to enlarged eyes. Here, a sonic hedgehog signaling activator increased retinal epithelia length in the organoid system when applied during but not after completion of development. This experimentally supports organoid maturation, stability, and experimental reproducibility in this organoid system, and provides a potential enlarged retina pathology model, as well as a protocol for producing larger organoids. Together, our study advances the understanding of retinal growth, maturation, and maintenance, and further optimizes the organoid system for future utilization.
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spelling pubmed-81140822021-05-13 Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture Völkner, Manuela Kurth, Thomas Schor, Jana Ebner, Lynn J. A. Bardtke, Lara Kavak, Cagri Hackermüller, Jörg Karl, Mike O. Front Cell Dev Biol Cell and Developmental Biology Using retinal organoid systems, organ-like 3D tissues, relies implicitly on their robustness. However, essential key parameters, particularly retinal growth and longer-term culture, are still insufficiently defined. Here, we hypothesize that a previously optimized protocol for high yield of evenly-sized mouse retinal organoids with low variability facilitates assessment of such parameters. We demonstrate that these organoids reliably complete retinogenesis, and can be maintained at least up to 60 days in culture. During this time, the organoids continue to mature on a molecular and (ultra)structural level: They develop photoreceptor outer segments and synapses, transiently maintain its cell composition for about 5–10 days after completing retinogenesis, and subsequently develop pathologic changes – mainly of the inner but also outer retina and reactive gliosis. To test whether this organoid system provides experimental access to the retina during and upon completion of development, we defined and stimulated organoid growth by activating sonic hedgehog signaling, which in patients and mice in vivo with a congenital defect leads to enlarged eyes. Here, a sonic hedgehog signaling activator increased retinal epithelia length in the organoid system when applied during but not after completion of development. This experimentally supports organoid maturation, stability, and experimental reproducibility in this organoid system, and provides a potential enlarged retina pathology model, as well as a protocol for producing larger organoids. Together, our study advances the understanding of retinal growth, maturation, and maintenance, and further optimizes the organoid system for future utilization. Frontiers Media S.A. 2021-04-27 /pmc/articles/PMC8114082/ /pubmed/33996806 http://dx.doi.org/10.3389/fcell.2021.645704 Text en Copyright © 2021 Völkner, Kurth, Schor, Ebner, Bardtke, Kavak, Hackermüller and Karl. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Völkner, Manuela
Kurth, Thomas
Schor, Jana
Ebner, Lynn J. A.
Bardtke, Lara
Kavak, Cagri
Hackermüller, Jörg
Karl, Mike O.
Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title_full Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title_fullStr Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title_full_unstemmed Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title_short Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture
title_sort mouse retinal organoid growth and maintenance in longer-term culture
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8114082/
https://www.ncbi.nlm.nih.gov/pubmed/33996806
http://dx.doi.org/10.3389/fcell.2021.645704
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