Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids
Objectives. Primary cilia are conserved organelles found in polarized mammalian cells that regulate neuronal growth, migration, and differentiation. Proper cilia formation is essential during eye development. Our previous reports found that both amacrine and retinal ganglion cells (RGCs) contain pri...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Hindawi Limited
2023-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2023/6494486 |
| _version_ | 1797945539481829376 |
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| author | Ke Ning Ziming Luo Tia J. Kowal Matthew Tran Rishab Majumder Trent M. Jarin Albert Y. Wu Jeffrey L. Goldberg Yang Sun |
| author_facet | Ke Ning Ziming Luo Tia J. Kowal Matthew Tran Rishab Majumder Trent M. Jarin Albert Y. Wu Jeffrey L. Goldberg Yang Sun |
| author_sort | Ke Ning |
| collection | DOAJ |
| description | Objectives. Primary cilia are conserved organelles found in polarized mammalian cells that regulate neuronal growth, migration, and differentiation. Proper cilia formation is essential during eye development. Our previous reports found that both amacrine and retinal ganglion cells (RGCs) contain primary cilia in primate and rodent retinas. However, whether primary cilia are present in the inner retina of human retinal organoids remains unknown. The purpose of this study is to characterize the primary cilia distribution in human embryonic stem cell (hESC-derived retinal organoid development. Materials and Methods. Retinal organoids were differentiated from a hESC line, harvested at various developmental timepoints (day 44-day 266), and immunostained with antibodies for primary cilia, including Arl13b (for the axoneme), AC3, and Centrin3 (for the basal body). AP2α, Prox1, GAD67, Calretinin, GFAP, PKCα, and Chx10 antibodies as well as Brn3b-promoted tdTomato expression were used to visualize retinal cell types. Results. A group of ciliated cells were present in the inner aspects of retinal organoids from day 44 to day 266 in culture. Ciliated Chx10-positive retinal progenitor cells, GFAP-positive astrocytes, and PKCα-positive rod-bipolar cells were detected later during development (day 176 to day 266). Ciliation persisted during all stages of retinal developmental in AP2α-positive amacrine cells, but it was decreased in Brn3b-positive retinal ganglion cells (RGCs) at later time points. Additionally, AC3-positive astrocytes significantly decreased during the later stages of organoid formation. Conclusions. Amacrine cells in retinal organoids retain cilia throughout development, whereas RGC ciliation gradually and progressively decreases with organoid maturation. |
| first_indexed | 2024-04-10T20:57:46Z |
| format | Article |
| id | doaj.art-69691f9dd4564113a4be480d77d35865 |
| institution | Directory Open Access Journal |
| issn | 1687-9678 |
| language | English |
| last_indexed | 2024-04-10T20:57:46Z |
| publishDate | 2023-01-01 |
| publisher | Hindawi Limited |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj.art-69691f9dd4564113a4be480d77d358652023-01-23T00:56:07ZengHindawi LimitedStem Cells International1687-96782023-01-01202310.1155/2023/6494486Characterization of Primary Cilia Formation in Human ESC-Derived Retinal OrganoidsKe Ning0Ziming Luo1Tia J. Kowal2Matthew Tran3Rishab Majumder4Trent M. Jarin5Albert Y. Wu6Jeffrey L. Goldberg7Yang Sun8Department of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyDepartment of OphthalmologyObjectives. Primary cilia are conserved organelles found in polarized mammalian cells that regulate neuronal growth, migration, and differentiation. Proper cilia formation is essential during eye development. Our previous reports found that both amacrine and retinal ganglion cells (RGCs) contain primary cilia in primate and rodent retinas. However, whether primary cilia are present in the inner retina of human retinal organoids remains unknown. The purpose of this study is to characterize the primary cilia distribution in human embryonic stem cell (hESC-derived retinal organoid development. Materials and Methods. Retinal organoids were differentiated from a hESC line, harvested at various developmental timepoints (day 44-day 266), and immunostained with antibodies for primary cilia, including Arl13b (for the axoneme), AC3, and Centrin3 (for the basal body). AP2α, Prox1, GAD67, Calretinin, GFAP, PKCα, and Chx10 antibodies as well as Brn3b-promoted tdTomato expression were used to visualize retinal cell types. Results. A group of ciliated cells were present in the inner aspects of retinal organoids from day 44 to day 266 in culture. Ciliated Chx10-positive retinal progenitor cells, GFAP-positive astrocytes, and PKCα-positive rod-bipolar cells were detected later during development (day 176 to day 266). Ciliation persisted during all stages of retinal developmental in AP2α-positive amacrine cells, but it was decreased in Brn3b-positive retinal ganglion cells (RGCs) at later time points. Additionally, AC3-positive astrocytes significantly decreased during the later stages of organoid formation. Conclusions. Amacrine cells in retinal organoids retain cilia throughout development, whereas RGC ciliation gradually and progressively decreases with organoid maturation.http://dx.doi.org/10.1155/2023/6494486 |
| spellingShingle | Ke Ning Ziming Luo Tia J. Kowal Matthew Tran Rishab Majumder Trent M. Jarin Albert Y. Wu Jeffrey L. Goldberg Yang Sun Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids Stem Cells International |
| title | Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids |
| title_full | Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids |
| title_fullStr | Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids |
| title_full_unstemmed | Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids |
| title_short | Characterization of Primary Cilia Formation in Human ESC-Derived Retinal Organoids |
| title_sort | characterization of primary cilia formation in human esc derived retinal organoids |
| url | http://dx.doi.org/10.1155/2023/6494486 |
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