Identification and Characterization of Human Retinal Stem-Like Cells with Regenerative Potential [snRNA-seq]

Human retinal stem cells (hRSCs) hold great promise in regenerative medicine due to their ability to directly differentiate into retinal cells. However, the characteristics and roles of hRSCs remain ambiguous. Here, we employed single-cell multi-omics profiles of human ciliary marginal zone (CMZ) of fetal retinas and human retinal organoids (hROs). We discovered a unique cell population of human retinal stem-like cells (hRSLCs) that specifically resides in the retinal tip and non-pigmented CMZ and possess a new regenerative lineage into neural retinal cells. Analysis of single cell and spatial transcriptomes show that the hRSLCs of hRO exhibit consistent characterization of cellular and molecular functions with fetal retinas. Functionally, several transcription factors were identified involved in the regulation of stemness from single cell RNA-seq and ATAC-seq data, and deficiency of gene MECOME in hRSLCs leds to impaired homeostasis and diminished retinal regeneration. Furthermore, transplantation of hRO-derived hRSLCs into retinal degeneration mice exhibit the high potential for retinal repair and improved visual function. Our work combines single cell multi-omics and organoid technologies to identify and comprehensively characterize a promising hRSLCs population in both human fetal retinas and organoid, opening up new avenues for potential therapy in retinal regeneration. Overall design: snRNA-seq for retinal organoids sample cultured for Day 60, Day 65, Day 70, Day 80, Day 90 and Day 100