Corneal organoids are useful tools for disease modeling and tissue transplantation; however, they have not yet been well studied during maturation. We characterized human iPSC-derived corneal organoids at 1, 2, 3, and 4 months of development using single-cell RNA sequencing to determine the cellular heterogeneity at each stage. We found pluripotent cell clusters committed to epithelial cell lineage at 1 month; early corneal epithelial, endothelial, and stromal cell markers at 2 months; keratocytes as the largest cell population at 3 months; and a large epithelial cell population at 4 months. We compared organoid to fetal corneal development at different stages and found that 4-month organoids closely resemble the corneal cellular complexity of the fetal (16 post conception week) and adult cornea. Using RNA velocity trajectory analysis, we found that less differentiated cells appear to give rise to corneal epithelial cells during development.

角膜类器官是疾病建模和组织移植的有用工具；然而，它们在成熟过程中尚未得到充分研究。我们使用单细胞 RNA 测序对人类 iPSC 衍生的角膜类器官在发育 1、2、3 和 4 个月时进行了表征，以确定每个阶段的细胞异质性。我们发现 1 个月时多能细胞簇致力于上皮细胞谱系；2 个月时早期角膜上皮、内皮和基质细胞标记物；角膜细胞是 3 个月时最大的细胞群；4个月时出现大量上皮细胞群。我们将不同阶段的类器官与胎儿角膜发育进行比较，发现 4 个月的类器官与胎儿（受孕后 16 周）和成人角膜的角膜细胞复杂性非常相似。使用RNA速度轨迹分析，我们发现分化程度较低的细胞似乎在发育过程中产生角膜上皮细胞。