Single-cell transcriptomic analysis of corneal organoids during development.

Area of Science:

• Ophthalmology
• Developmental Biology
• Stem Cell Biology

Background:

• Corneal organoids are valuable for disease modeling and transplantation.
• Their developmental maturation process remains incompletely understood.

Purpose of the Study:

• To characterize the cellular heterogeneity of human induced pluripotent stem cell (iPSC)-derived corneal organoids during maturation.
• To compare organoid development with fetal corneal development.

Main Methods:

• Single-cell RNA sequencing was employed to analyze corneal organoids at 1, 2, 3, and 4 months of development.
• RNA velocity trajectory analysis was used to infer cell differentiation pathways.

Main Results:

• Pluripotent cell clusters committed to the epithelial lineage were observed at 1 month.
• Early markers for corneal epithelial, endothelial, and stromal cells appeared at 2 months.
• Keratocytes became the predominant cell type by 3 months, followed by a dominant epithelial cell population at 4 months.
• Four-month-old organoids exhibited cellular complexity comparable to fetal (16 weeks post-conception) and adult corneas.
• RNA velocity analysis indicated that less differentiated cells give rise to corneal epithelial cells.

Conclusions:

• Human iPSC-derived corneal organoids recapitulate key stages of corneal development.
• Mature corneal organoids serve as a robust model for studying corneal biology and disease.
• This study provides insights into the differentiation dynamics of corneal cell types within organoids.