Suppr超能文献

长期培养中小鼠视网膜类器官的生长与维持

Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture.

作者信息

Völkner Manuela, Kurth Thomas, Schor Jana, Ebner Lynn J A, Bardtke Lara, Kavak Cagri, Hackermüller Jörg, Karl Mike O

机构信息

German Center for Neurodegenerative Diseases (DZNE) Dresden, Dresden, Germany.

Center for Molecular and Cellular Bioengineering, Technology Platform, Electron Microscopy and Histology Facility, Technische Universität Dresden, Dresden, Germany.

出版信息

Front Cell Dev Biol. 2021 Apr 27;9:645704. doi: 10.3389/fcell.2021.645704. eCollection 2021.

Abstract

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 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.

摘要

利用视网膜类器官系统,即类器官样三维组织,隐含地依赖于它们的稳健性。然而,关键参数,尤其是视网膜生长和长期培养,仍未得到充分界定。在此,我们假设,一种先前优化的方案,用于高产、大小均匀且变异性低的小鼠视网膜类器官,有助于评估此类参数。我们证明,这些类器官能可靠地完成视网膜发育,并且在培养中至少可维持60天。在此期间,类器官在分子和(超)结构水平上持续成熟:它们发育出光感受器外段和突触,在完成视网膜发育后短暂维持其细胞组成约5 - 10天,随后出现病理变化——主要在内层视网膜,但外层视网膜和反应性胶质增生也有变化。为了测试这个类器官系统在发育期间及发育完成后是否能提供对视网膜的实验性研究途径,我们通过激活音猬因子信号通路来定义和刺激类器官生长,在患有先天性缺陷的患者和小鼠中,该信号通路会导致眼睛增大。在此,当在发育期间而非发育完成后应用时,音猬因子信号通路激活剂增加了类器官系统中视网膜上皮的长度。这在实验上支持了该类器官系统中的类器官成熟、稳定性和实验可重复性,并提供了一个潜在的扩大视网膜病理模型,以及一种生产更大类器官的方案。总之,我们的研究推进了对视网膜生长、成熟和维持的理解,并进一步优化了类器官系统以供未来使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d2f/8114082/40aa1c2dfd2b/fcell-09-645704-g001.jpg

相似文献

1
Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture.
Front Cell Dev Biol. 2021 Apr 27;9:645704. doi: 10.3389/fcell.2021.645704. eCollection 2021.
2
Hyaluronan improves photoreceptor differentiation and maturation in human retinal organoids.
Acta Biomater. 2024 Jun;181:117-132. doi: 10.1016/j.actbio.2024.05.001. Epub 2024 May 3.
7
Retinal Organoids from Pluripotent Stem Cells Efficiently Recapitulate Retinogenesis.
Stem Cell Reports. 2016 Apr 12;6(4):525-538. doi: 10.1016/j.stemcr.2016.03.001. Epub 2016 Mar 31.
8
Structural and Functional Characterization of Human Stem-Cell-Derived Retinal Organoids by Live Imaging.
Invest Ophthalmol Vis Sci. 2017 Jul 1;58(9):3311-3318. doi: 10.1167/iovs.16-20796.
9
The Analysis of Embryoid Body Formation and Its Role in Retinal Organoid Development.
Int J Mol Sci. 2024 Jan 24;25(3):1444. doi: 10.3390/ijms25031444.
10
From Otic Induction to Hair Cell Production: Pax2 Cell Line Illuminates Key Stages of Development in Mouse Inner Ear Organoid Model.
Stem Cells Dev. 2018 Feb 15;27(4):237-251. doi: 10.1089/scd.2017.0142. Epub 2018 Jan 29.

引用本文的文献

1
Bridging the gap: the role of 3D cell cultures in mimicking tumor microenvironment for enhanced drug testing accuracy.
Front Bioeng Biotechnol. 2025 Aug 12;13:1498141. doi: 10.3389/fbioe.2025.1498141. eCollection 2025.
2
Characterization of age-related dolichol increases in the mouse retina.
Res Sq. 2025 May 16:rs.3.rs-6498277. doi: 10.21203/rs.3.rs-6498277/v1.
3
Robust generation of photoreceptor-dominant retinal organoids from porcine induced pluripotent stem cells.
Stem Cell Reports. 2025 Apr 8;20(4):102425. doi: 10.1016/j.stemcr.2025.102425. Epub 2025 Mar 6.
4
DNA Origami Barcodes for Immunostaining.
ACS Appl Mater Interfaces. 2025 Mar 12;17(10):15813-15823. doi: 10.1021/acsami.4c19153. Epub 2025 Feb 27.
5
Automated quantification of photoreceptor outer segments in developing and degenerating retinas on microscopy images across scales.
Front Mol Neurosci. 2024 May 24;17:1398447. doi: 10.3389/fnmol.2024.1398447. eCollection 2024.
6
Intrinsic signal optoretinography of dark adaptation abnormality due to rod photoreceptor degeneration.
Exp Biol Med (Maywood). 2024 Jan 31;249:10024. doi: 10.3389/ebm.2024.10024. eCollection 2024.
7
Deletion of IFT20 exclusively in the RPE ablates primary cilia and leads to retinal degeneration.
PLoS Biol. 2023 Dec 4;21(12):e3002402. doi: 10.1371/journal.pbio.3002402. eCollection 2023 Dec.
8
Reliability of human retina organoid generation from hiPSC-derived neuroepithelial cysts.
Front Cell Neurosci. 2023 Oct 6;17:1166641. doi: 10.3389/fncel.2023.1166641. eCollection 2023.
9
Mathematical model for glutathione dynamics in the retina.
Sci Rep. 2023 Jul 7;13(1):10996. doi: 10.1038/s41598-023-37938-9.
10
Modeling inducible neuropathologies of the retina with differential phenotypes in organoids.
Front Cell Neurosci. 2023 May 5;17:1106287. doi: 10.3389/fncel.2023.1106287. eCollection 2023.

本文引用的文献

1
Retinal organoids: a window into human retinal development.
Development. 2020 Dec 24;147(24):dev189746. doi: 10.1242/dev.189746.
2
Human Organoids for the Study of Retinal Development and Disease.
Annu Rev Vis Sci. 2020 Sep 15;6:91-114. doi: 10.1146/annurev-vision-121219-081855.
3
Hydrogel-based milliwell arrays for standardized and scalable retinal organoid cultures.
Sci Rep. 2020 Jun 24;10(1):10275. doi: 10.1038/s41598-020-67012-7.
5
Modeling and Rescue of RP2 Retinitis Pigmentosa Using iPSC-Derived Retinal Organoids.
Stem Cell Reports. 2020 Jul 14;15(1):67-79. doi: 10.1016/j.stemcr.2020.05.007. Epub 2020 Jun 11.
6
Pluripotent stem cell-derived retinal organoids for disease modeling and development of therapies.
Stem Cells. 2020 Oct 1;38(10):1206-1215. doi: 10.1002/stem.3239. Epub 2020 Jun 7.
8
uap: reproducible and robust HTS data analysis.
BMC Bioinformatics. 2019 Dec 12;20(1):664. doi: 10.1186/s12859-019-3219-1.
10
Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis.
Stem Cell Reports. 2019 Nov 12;13(5):906-923. doi: 10.1016/j.stemcr.2019.09.010. Epub 2019 Oct 24.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验