Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors
Three-dimensional retinal culture systems help to understand eye development and the pathology of disorders. There is a need for reporter stem cell lines to allow in vitro studies on retinal progenitors and photoreceptors and their developmental dynamics or properties and to test therapeutic approac...
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MDPI AG
2025-02-01
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| Online Access: | https://www.mdpi.com/2073-4409/14/4/252 |
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| author | Oleksandr Zabiegalov Adeline Berger Dhryata Kamdar Kabirou Adamou Chuanxi Tian Martial Mbefo Mathieu Quinodoz Florian Udry Carlo Rivolta Corinne Kostic Yvan Arsenijevic |
| author_facet | Oleksandr Zabiegalov Adeline Berger Dhryata Kamdar Kabirou Adamou Chuanxi Tian Martial Mbefo Mathieu Quinodoz Florian Udry Carlo Rivolta Corinne Kostic Yvan Arsenijevic |
| author_sort | Oleksandr Zabiegalov |
| collection | DOAJ |
| description | Three-dimensional retinal culture systems help to understand eye development and the pathology of disorders. There is a need for reporter stem cell lines to allow in vitro studies on retinal progenitors and photoreceptors and their developmental dynamics or properties and to test therapeutic approaches. The isolation of pure progenitor populations or photoreceptor precursors may serve for drug, gene, and cell therapy development. Here, we generated a dual-reporter mouse embryonic stem cell line <i>Crx-GFP;Rax-mCherry</i> enabling the visualization or isolation of photoreceptors and retinal progenitors from retinal organoid settings. From day 4 organoids, we isolated mCherry-positive cells to assess their early retinal progenitor identity with proliferation tests as well as transcriptomic and proteomic profiling. The timing of eye field transcription factor expression at the transcriptomic and protein levels is in accordance with mouse retinogenesis. This new line will be helpful for rapidly investigating biological questions or testing therapeutics before using human induced pluripotent stem cells (iPSCs), which require a much longer time for retinal organoid formation. |
| format | Article |
| id | doaj-art-4a7871bb230042e4943548dade1d9368 |
| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj-art-4a7871bb230042e4943548dade1d93682025-08-20T02:44:59ZengMDPI AGCells2073-44092025-02-0114425210.3390/cells14040252Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and PhotoreceptorsOleksandr Zabiegalov0Adeline Berger1Dhryata Kamdar2Kabirou Adamou3Chuanxi Tian4Martial Mbefo5Mathieu Quinodoz6Florian Udry7Carlo Rivolta8Corinne Kostic9Yvan Arsenijevic10Unit of Retinal Degeneration and Regeneration, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandUnit of Epigenetics of Ocular Diseases, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandOphthalmic Genetics Group, Institute of Molecular and Clinical Ophthalmology, 4031 Basel, SwitzerlandGroup for Retinal Disorders Research, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandUnit of Retinal Degeneration and Regeneration, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandUnit of Retinal Degeneration and Regeneration, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandOphthalmic Genetics Group, Institute of Molecular and Clinical Ophthalmology, 4031 Basel, SwitzerlandUnit of Retinal Degeneration and Regeneration, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandOphthalmic Genetics Group, Institute of Molecular and Clinical Ophthalmology, 4031 Basel, SwitzerlandGroup for Retinal Disorders Research, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandUnit of Retinal Degeneration and Regeneration, Department of Ophthalmology, University of Lausanne, 1004 Lausanne, SwitzerlandThree-dimensional retinal culture systems help to understand eye development and the pathology of disorders. There is a need for reporter stem cell lines to allow in vitro studies on retinal progenitors and photoreceptors and their developmental dynamics or properties and to test therapeutic approaches. The isolation of pure progenitor populations or photoreceptor precursors may serve for drug, gene, and cell therapy development. Here, we generated a dual-reporter mouse embryonic stem cell line <i>Crx-GFP;Rax-mCherry</i> enabling the visualization or isolation of photoreceptors and retinal progenitors from retinal organoid settings. From day 4 organoids, we isolated mCherry-positive cells to assess their early retinal progenitor identity with proliferation tests as well as transcriptomic and proteomic profiling. The timing of eye field transcription factor expression at the transcriptomic and protein levels is in accordance with mouse retinogenesis. This new line will be helpful for rapidly investigating biological questions or testing therapeutics before using human induced pluripotent stem cells (iPSCs), which require a much longer time for retinal organoid formation.https://www.mdpi.com/2073-4409/14/4/252retinal organoidsreporter linestem cellsretinal progenitor cells<i>Rax</i> genecell tracing |
| spellingShingle | Oleksandr Zabiegalov Adeline Berger Dhryata Kamdar Kabirou Adamou Chuanxi Tian Martial Mbefo Mathieu Quinodoz Florian Udry Carlo Rivolta Corinne Kostic Yvan Arsenijevic Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors Cells retinal organoids reporter line stem cells retinal progenitor cells <i>Rax</i> gene cell tracing |
| title | Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors |
| title_full | Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors |
| title_fullStr | Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors |
| title_full_unstemmed | Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors |
| title_short | Generation of a Double Reporter mES Cell Line to Simultaneously Trace the Generation of Retinal Progenitors and Photoreceptors |
| title_sort | generation of a double reporter mes cell line to simultaneously trace the generation of retinal progenitors and photoreceptors |
| topic | retinal organoids reporter line stem cells retinal progenitor cells <i>Rax</i> gene cell tracing |
| url | https://www.mdpi.com/2073-4409/14/4/252 |
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