Cargando…

Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids

Retinal organoids (ROs) are three-dimensional retinal tissues, which are differentiated in vitro from induced pluripotent stem cells (iPSC), ultimately forming all main retinal cell types under defined culture conditions. ROs show several highly specialized retinal features, including the outgrowth...

Descripción completa

Detalles Bibliográficos
Autores principales: Berber, Patricia, Bondarenko, Sofiia, Michaelis, Lisa, Weber, Bernhard Heinrich Friedrich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9739155/
https://www.ncbi.nlm.nih.gov/pubmed/36499228
http://dx.doi.org/10.3390/ijms232314893
_version_ 1784847734947184640
author Berber, Patricia
Bondarenko, Sofiia
Michaelis, Lisa
Weber, Bernhard Heinrich Friedrich
author_facet Berber, Patricia
Bondarenko, Sofiia
Michaelis, Lisa
Weber, Bernhard Heinrich Friedrich
author_sort Berber, Patricia
collection PubMed
description Retinal organoids (ROs) are three-dimensional retinal tissues, which are differentiated in vitro from induced pluripotent stem cells (iPSC), ultimately forming all main retinal cell types under defined culture conditions. ROs show several highly specialized retinal features, including the outgrowth of photoreceptor outer segments (OSs). In vivo, the photoreceptor OSs are enveloped and maintained by protrusions of retinal pigment epithelium (RPE) cells, the so-called apical microvilli, while ROs fail to recapitulate this critical interaction in culture development. Here, we define specific co-culture conditions aiming to compensate for the missing physical proximity of RPE and OSs in RO development. Accordingly, functional RPE cells and ROs were differentiated simultaneously from the same iPSC clone, the former resulting in byproduct RPE or bRPE cells. While some co-culture approaches indicated a temporary functional interaction between bRPE and RO photoreceptors, they did not improve the photoreceptor histoarchitecture. In contrast, embedding ROs in a basement membrane extract without bRPE cells showed a robust improvement in the rate of photoreceptor OS retention. RO embedding is a quick and easy method that greatly enhances the preservation of photoreceptor OSs, an important structure for modelling retinal diseases with the involvement of photoreceptors.
format Online
Article
Text
id pubmed-9739155
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-97391552022-12-11 Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids Berber, Patricia Bondarenko, Sofiia Michaelis, Lisa Weber, Bernhard Heinrich Friedrich Int J Mol Sci Article Retinal organoids (ROs) are three-dimensional retinal tissues, which are differentiated in vitro from induced pluripotent stem cells (iPSC), ultimately forming all main retinal cell types under defined culture conditions. ROs show several highly specialized retinal features, including the outgrowth of photoreceptor outer segments (OSs). In vivo, the photoreceptor OSs are enveloped and maintained by protrusions of retinal pigment epithelium (RPE) cells, the so-called apical microvilli, while ROs fail to recapitulate this critical interaction in culture development. Here, we define specific co-culture conditions aiming to compensate for the missing physical proximity of RPE and OSs in RO development. Accordingly, functional RPE cells and ROs were differentiated simultaneously from the same iPSC clone, the former resulting in byproduct RPE or bRPE cells. While some co-culture approaches indicated a temporary functional interaction between bRPE and RO photoreceptors, they did not improve the photoreceptor histoarchitecture. In contrast, embedding ROs in a basement membrane extract without bRPE cells showed a robust improvement in the rate of photoreceptor OS retention. RO embedding is a quick and easy method that greatly enhances the preservation of photoreceptor OSs, an important structure for modelling retinal diseases with the involvement of photoreceptors. MDPI 2022-11-28 /pmc/articles/PMC9739155/ /pubmed/36499228 http://dx.doi.org/10.3390/ijms232314893 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Berber, Patricia
Bondarenko, Sofiia
Michaelis, Lisa
Weber, Bernhard Heinrich Friedrich
Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title_full Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title_fullStr Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title_full_unstemmed Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title_short Transient Retention of Photoreceptor Outer Segments in Matrigel-Embedded Retinal Organoids
title_sort transient retention of photoreceptor outer segments in matrigel-embedded retinal organoids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9739155/
https://www.ncbi.nlm.nih.gov/pubmed/36499228
http://dx.doi.org/10.3390/ijms232314893
work_keys_str_mv AT berberpatricia transientretentionofphotoreceptoroutersegmentsinmatrigelembeddedretinalorganoids
AT bondarenkosofiia transientretentionofphotoreceptoroutersegmentsinmatrigelembeddedretinalorganoids
AT michaelislisa transientretentionofphotoreceptoroutersegmentsinmatrigelembeddedretinalorganoids
AT weberbernhardheinrichfriedrich transientretentionofphotoreceptoroutersegmentsinmatrigelembeddedretinalorganoids