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Putative Self-Organizing Human Corneal Organoids Recapitulate Human Corneal Architecture and Cellular Diversity
Timothy A Blenkinsop1, Anne Z Eriksen2
1Department of Ophthalmology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Bioengineering (Basel, Switzerland)
|May 27, 2026
Summary
Researchers developed a novel 3D corneal organoid model from human embryonic stem cells (hESCs) that successfully mimics the complex structure and cell types of the human cornea for disease research.
Area of Science:
- Stem cell biology
- Ophthalmology
- Tissue engineering
Background:
- Pluripotent stem cell-derived corneal organoids are valuable for studying corneal development and disease.
- Existing models require improvement to fully replicate native human cornea's complexity.
- A need exists for advanced 3D models recapitulating corneal architecture and cellular diversity.
Purpose of the Study:
- To develop a modified spontaneous 3D corneal organoid model using hESCs.
- To create a more accurate in vitro model of human corneal tissue.
- To utilize an adapted Self-formed Ectoderm Autonomous Multi-zone (SEAM) protocol.
Main Methods:
- Human embryonic stem cells (hESCs) were cultured as spheroids and differentiated for 7-8 weeks.
- Organoids were analyzed using immunofluorescence staining for corneal markers.
- Single-cell RNA sequencing (scRNA-seq) was employed to determine cellular composition and gene expression.
Main Results:
- Approximately 20% of organoids exhibited transparent corneal regions by day 30.
- Immunofluorescence confirmed spatially organized corneal markers (ZO-1, E-cadherin, P63α, vimentin, laminin-1).
- scRNA-seq identified 18 cell clusters, including distinct epithelial, stromal, and endothelial populations, with key corneal gene expression.
Conclusions:
- The 3D spheroid-based corneal organoid model recapitulates human cornea's multilayered architecture and cellular diversity.
- The model exhibits molecular signatures consistent with native corneal tissue.
- This platform is valuable for studying corneal development, disease mechanisms, and therapeutic applications.
