Laboratory for Retinal Regeneration, RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo 650-0047, Japan; Department of Ophthalmology, Kobe City Eye Hospital, Kobe, Hyogo 650-0047, Japan; Cell and Gene Therapy in Ophthalmology Laboratory, BZP, RIKEN, Wako, Saitama 351-0198, Japan; Department of Ophthalmology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
ThinkCyte K.K., Tokyo 113-8654, Japan.
Stem Cell Reports. 2024 Feb 13;19(2):254-269. doi: 10.1016/j.stemcr.2023.12.001. Epub 2024 Jan 4.
Pluripotent stem cell-based therapy for retinal degenerative diseases is a promising approach to restoring visual function. A clinical study using retinal organoid (RO) sheets was recently conducted in patients with retinitis pigmentosa. However, the graft preparation currently requires advanced skills to identify and excise suitable segments from the transplantable area of the limited number of suitable ROs. This remains a challenge for consistent clinical implementations. Herein, we enabled the enrichment of wild-type (non-reporter) retinal progenitor cells (RPCs) from dissociated ROs using a label-free ghost cytometry (LF-GC)-based sorting system, where a machine-based classifier was trained in advance with another RPC reporter line. The sorted cells reproducibly formed retinal spheroids large enough for transplantation and developed mature photoreceptors in the retinal degeneration rats. This method of enriching early RPCs with no specific surface antigens and without any reporters or chemical labeling is promising for robust preparation of graft tissues during cell-based therapy.
基于多能干细胞的视网膜退行性疾病治疗方法是恢复视觉功能的一种很有前途的方法。最近，一项使用视网膜类器官（RO）片的临床研究在色素性视网膜炎患者中进行。然而，目前的移植物制备需要先进的技能，以从有限数量的合适 RO 中移植区域中识别和切除合适的片段。这对于一致的临床实施仍然是一个挑战。在这里，我们使用无标记鬼细胞术（LF-GC）为基础的分选系统，从分离的 RO 中富集野生型（非报告）视网膜祖细胞（RPC），其中机器基分类器在另一个 RPC 报告细胞系中提前进行了训练。分选后的细胞可重复性地形成足够大的视网膜球体，用于移植，并在视网膜变性大鼠中发育成熟的光感受器。这种没有特定表面抗原的早期 RPC 富集方法，并且没有任何报告基因或化学标记，有望在基于细胞的治疗中稳健地制备移植物组织。
