Moosajee, M;
Eintracht, J;
Harding, P;
Lima Cunha, D;
(2022)
Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells [version 1; peer review: 2 approved].
F1000Research
, 11
, Article 324. 10.12688/f1000research.108829.1.
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Abstract
Animal models have provided many insights into ocular development and disease, but they remain suboptimal for understanding human oculogenesis. Eye development requires spatiotemporal gene expression patterns and disease phenotypes can differ significantly between humans and animal models, with patient-associated mutations causing embryonic lethality reported in some animal models. The emergence of human induced pluripotent stem cell (hiPSC) technology has provided a new resource for dissecting the complex nature of early eye morphogenesis through the generation of three-dimensional (3D) cellular models. By using patient-specific hiPSCs to generate in vitro optic vesicle-like models, we can enhance the understanding of early developmental eye disorders and provide a pre-clinical platform for disease modelling and therapeutics testing. A major challenge of in vitro optic vesicle generation is the low efficiency of differentiation in 3D cultures. To address this, we adapted a previously published protocol of retinal organoid differentiation to improve embryoid body formation using a microwell plate. Established morphology, upregulated transcript levels of known early eye-field transcription factors and protein expression of standard retinal progenitor markers confirmed the optic vesicle/presumptive optic cup identity of in vitro models between day 20 and 50 of culture. This adapted protocol is relevant to researchers seeking a physiologically relevant model of early human ocular development and disease with a view to replacing animal models.
| Type: | Article |
|---|---|
| Title: | Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells [version 1; peer review: 2 approved] |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.12688/f1000research.108829.1 |
| Publisher version: | https://doi.org/10.12688/f1000research.108829.1 |
| Language: | English |
| Additional information: | © 2022 Eintracht J et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Embryoid bodies, PAX6, VSX2, eye development, iPSCs, optic vesicles, retinal differentiation, Animals, Cell Differentiation, Embryoid Bodies, Humans, Induced Pluripotent Stem Cells, Retina, Transcription Factors |
| UCL classification: | UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10152642 |
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