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Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid
BACKGROUND: Human-induced pluripotent stem cell-derived retinal organoids are a valuable tool for disease modelling and therapeutic development. Many efforts have been made over the last decade to optimise protocols for the generation of organoids that correctly mimic the human retina. Most protocol...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9482314/ https://www.ncbi.nlm.nih.gov/pubmed/36114559 http://dx.doi.org/10.1186/s13287-022-03146-x |
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author | Sanjurjo-Soriano, Carla Erkilic, Nejla Damodar, Krishna Boukhaddaoui, Hassan Diakatou, Michalitsa Garita-Hernandez, Marcela Mamaeva, Daria Dubois, Gregor Jazouli, Zhour Jimenez-Medina, Carla Goureau, Olivier Meunier, Isabelle Kalatzis, Vasiliki |
author_facet | Sanjurjo-Soriano, Carla Erkilic, Nejla Damodar, Krishna Boukhaddaoui, Hassan Diakatou, Michalitsa Garita-Hernandez, Marcela Mamaeva, Daria Dubois, Gregor Jazouli, Zhour Jimenez-Medina, Carla Goureau, Olivier Meunier, Isabelle Kalatzis, Vasiliki |
author_sort | Sanjurjo-Soriano, Carla |
collection | PubMed |
description | BACKGROUND: Human-induced pluripotent stem cell-derived retinal organoids are a valuable tool for disease modelling and therapeutic development. Many efforts have been made over the last decade to optimise protocols for the generation of organoids that correctly mimic the human retina. Most protocols use common media supplements; however, protocol-dependent variability impacts data interpretation. To date, the lack of a systematic comparison of a given protocol with or without supplements makes it difficult to determine how they influence the differentiation process and morphology of the retinal organoids. METHODS: A 2D-3D differentiation method was used to generate retinal organoids, which were cultured with or without the most commonly used media supplements, notably retinoic acid. Gene expression was assayed using qPCR analysis, protein expression using immunofluorescence studies, ultrastructure using electron microscopy and 3D morphology using confocal and biphoton microscopy of whole organoids. RESULTS: Retinoic acid delayed the initial stages of differentiation by modulating photoreceptor gene expression. At later stages, the presence of retinoic acid led to the generation of mature retinal organoids with a well-structured stratified photoreceptor layer containing a predominant rod population. By contrast, the absence of retinoic acid led to cone-rich organoids with a less organised and non-stratified photoreceptor layer. CONCLUSIONS: This study proves the importance of supplemented media for culturing retinal organoids. More importantly, we demonstrate for the first time that the role of retinoic acid goes beyond inducing a rod cell fate to enhancing the organisation of the photoreceptor layer of the mature organoid. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-03146-x. |
format | Online Article Text |
id | pubmed-9482314 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-94823142022-09-18 Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid Sanjurjo-Soriano, Carla Erkilic, Nejla Damodar, Krishna Boukhaddaoui, Hassan Diakatou, Michalitsa Garita-Hernandez, Marcela Mamaeva, Daria Dubois, Gregor Jazouli, Zhour Jimenez-Medina, Carla Goureau, Olivier Meunier, Isabelle Kalatzis, Vasiliki Stem Cell Res Ther Research BACKGROUND: Human-induced pluripotent stem cell-derived retinal organoids are a valuable tool for disease modelling and therapeutic development. Many efforts have been made over the last decade to optimise protocols for the generation of organoids that correctly mimic the human retina. Most protocols use common media supplements; however, protocol-dependent variability impacts data interpretation. To date, the lack of a systematic comparison of a given protocol with or without supplements makes it difficult to determine how they influence the differentiation process and morphology of the retinal organoids. METHODS: A 2D-3D differentiation method was used to generate retinal organoids, which were cultured with or without the most commonly used media supplements, notably retinoic acid. Gene expression was assayed using qPCR analysis, protein expression using immunofluorescence studies, ultrastructure using electron microscopy and 3D morphology using confocal and biphoton microscopy of whole organoids. RESULTS: Retinoic acid delayed the initial stages of differentiation by modulating photoreceptor gene expression. At later stages, the presence of retinoic acid led to the generation of mature retinal organoids with a well-structured stratified photoreceptor layer containing a predominant rod population. By contrast, the absence of retinoic acid led to cone-rich organoids with a less organised and non-stratified photoreceptor layer. CONCLUSIONS: This study proves the importance of supplemented media for culturing retinal organoids. More importantly, we demonstrate for the first time that the role of retinoic acid goes beyond inducing a rod cell fate to enhancing the organisation of the photoreceptor layer of the mature organoid. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-03146-x. BioMed Central 2022-09-16 /pmc/articles/PMC9482314/ /pubmed/36114559 http://dx.doi.org/10.1186/s13287-022-03146-x Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Sanjurjo-Soriano, Carla Erkilic, Nejla Damodar, Krishna Boukhaddaoui, Hassan Diakatou, Michalitsa Garita-Hernandez, Marcela Mamaeva, Daria Dubois, Gregor Jazouli, Zhour Jimenez-Medina, Carla Goureau, Olivier Meunier, Isabelle Kalatzis, Vasiliki Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title | Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title_full | Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title_fullStr | Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title_full_unstemmed | Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title_short | Retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
title_sort | retinoic acid delays initial photoreceptor differentiation and results in a highly structured mature retinal organoid |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9482314/ https://www.ncbi.nlm.nih.gov/pubmed/36114559 http://dx.doi.org/10.1186/s13287-022-03146-x |
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