Stone Nicholas E, Bohrer Laura R, Mullin Nathaniel K, Berthold Alexander, Wright Allison T, Han Ian C, Stone Edwin M, Mullins Robert F, Tucker Budd A
Institute for Vision Research.
Department of Ophthalmology and Visual Sciences, and.
JCI Insight. 2025 Jun 12;10(14). doi: 10.1172/jci.insight.186338. eCollection 2025 Jul 22.
Autologous photoreceptor cell replacement is one of the most promising strategies currently being developed for the treatment of patients with inherited retinal degenerative blindness. Induced pluripotent stem cell-derived (iPSC-derived) retinal organoids, which faithfully recapitulate the structure of the neural retina, are an ideal source of transplantable photoreceptors required for these therapies. However, retinal organoids contain other retinal cell types, including bipolar, horizontal, and amacrine cells, which are unneeded and may reduce the potency of the final therapeutic product. Therefore, approaches for isolating fate-committed photoreceptor cells from dissociated retinal organoids are desirable. In this work, we present partial dissociation, a technique that leverages the high level of organization found in retinal organoids to enable selective enrichment of photoreceptor cells without the use of specialized equipment or reagents such as antibody labels. We demonstrate up to 90% photoreceptor cell purity by simply selecting cell fractions liberated from retinal organoids during enzymatic digestion in the absence of mechanical dissociation. Since the presented approach relies on the use of standard plasticware and commercially available current good manufacturing practice-compliant reagents, we believe that it is ideal for use in the preparation of clinical photoreceptor cell replacement therapies.
自体光感受器细胞替代是目前正在开发的用于治疗遗传性视网膜退行性失明患者最有前景的策略之一。诱导多能干细胞衍生的（iPSC衍生的）视网膜类器官能够忠实地重现神经视网膜的结构，是这些疗法所需的可移植光感受器的理想来源。然而，视网膜类器官包含其他视网膜细胞类型，包括双极细胞、水平细胞和无长突细胞，这些细胞是不需要的，并且可能会降低最终治疗产品的效力。因此，需要从解离的视网膜类器官中分离已确定命运的光感受器细胞的方法。在这项工作中，我们提出了部分解离技术，该技术利用视网膜类器官中发现的高度组织化，在不使用诸如抗体标记等专门设备或试剂的情况下，实现光感受器细胞的选择性富集。我们通过在没有机械解离的情况下，简单地选择酶消化过程中从视网膜类器官中释放的细胞部分，证明了高达90%的光感受器细胞纯度。由于所提出的方法依赖于使用标准塑料制品和符合现行良好生产规范的市售试剂，我们认为它非常适合用于临床光感受器细胞替代疗法的制备。
