Protocol for modeling injury-induced regeneration in retinal organoids and isolating hNRSCs for subretinal transplantation in rd10 mice
Human retinal organoids (hROs) provide a platform for modeling injury and assessing regenerative capacity in vitro. Here, we present a protocol for stepwise generation of hROs from human embryonic stem cells (hESCs). We describe steps for microsurgical induction of neural retina (NR) injury, isolation of CPAMD8<sup>+</sup> human neural retinal stem-like cells (hNRSCs) using flow cytometry, and their subretinal transplantation into rd10 mice. By isolating and subretinally injecting hESC-derived neural retinal stem-like cells, this protocol enables subsequent functional studies in retinal degeneration models. For complete details on the use and execution of this protocol, please refer to Liu et al.<sup>1</sup>.
Authors
- Cheng Chen (ORCID: https://orcid.org/0000-0003-1065-7201)
- Na Gao
- Weisan Pan
- Jianzhong Su (ORCID: https://orcid.org/0000-0002-1172-7411)
- Shasha Li (ORCID: https://orcid.org/0000-0003-3823-3419)
- Hui Liu (ORCID: https://orcid.org/0000-0003-0444-5579)
- Ming Chen (ORCID: https://orcid.org/0009-0009-1087-4186)
- Xiaoyu Liu (ORCID: https://orcid.org/0009-0006-9888-4773)
- Yingfan Zhang (ORCID: https://orcid.org/0000-0002-5002-6896)
Institutions
- Wenzhou Medical University (CN)
- Affiliated Eye Hospital of Wenzhou Medical College (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-11-27
- DOI
- https://doi.org/10.1016/j.xpro.2025.104232
- Primary Topic
- Retinal Development and Disorders
- Type
- article
- Field-Weighted Citation Impact
- 0.00