Mouse pluripotent stem cells can be efficiently differentiated into retinal organoids with polarized, laminated neural retina harboring all retinal cell types by the Hypoxia-Induced Generation of Photoreceptor in Retinal Organoids (HIPRO) protocol. In our recent publication, we modified the HIPRO protocol on the basis of comparative transcriptome analyses to facilitate photoreceptor biogenesis and maturation. Here, we provide a detailed protocol for efficient generation of retinal organoids from mouse pluripotent stem cells. For complete details on the use and execution of this protocol, please refer to (Chen et al., 2016, DiStefano et al., 2018, Brooks et al., 2019).
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7402619 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2020.100018 | DOI Listing |
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- Histology is crucial for examining tissue structure and cell details, but standard methods for cryosectioning small tissues like organoids lack efficiency and cost-effectiveness, hindering analysis.
- The adapted HistoBrick method uses an optimal embedding mixture of 8% PEGDA and 2.5% gelatine, providing support for fragile samples during cryosectioning and preserving delicate structures of human retinal organoids.
- Using these PEGDA-gelatine HistoBricks, researchers monitored retinal organoid development over time, finding that photoreceptor cell bodies were sustained for up to 98 weeks, although outer segments diminished, making this approach valuable for increased throughput in tissue studies and research.
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