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Updated: Sep 9, 2025
A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice
Published on: December 18, 2015
Single-cell RNA sequencing of mouse cornea during wound healing after infrared laser irradiation
Yixue Yin1, Luguang Jiao2, Jiarui Wang2
1Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine, Jinan, 250002, China; School of Basic Medical Sciences, Anhui Medical University, Hefei, 230032, China.
Abstract:
The cornea is highly specialized and a detailed map of the diverse component cell types is a vital tool for ocular disease treatment and corneal healing. Single-cell RNA sequencing (scRNA-seq) was performed to analyze corneal wound healing in C57BL/6J mice following 3.74 μm infrared laser injury (23.2 J/cm2). Analysis of 12,956 cells from 10 healthy corneas identified 28 clusters of epithelial stem cells, differentiated epithelial cells, stromal cells, endothelial cells, immune cells, Schwann cells and progenitor populations. Three newly differentiated epithelial subpopulations emerged at 3 days post-injury and a marked increase in immune cells, especially neutrophils and CD163+ macrophages, was seen. Differentiated epithelial cells and normal stromal cells decreased, myofibroblasts and stromal stem cells increased and endothelial cells and Schwann cells declined. Immune cell numbers and myofibroblasts had decreased by 21 days post-injury and differentiated epithelial cells, normal stromal cells and stromal stem cells had increased. Endothelial cells and Schwann cells showed partial recovery. Corneal repair was composed of two distinct phases. An early inflammatory phase was characterized by immune activation and epithelial remodeling and a late recovery phase by stromal restoration, myofibroblast regression and neural/endothelial regeneration. Dynamic cellular interactions during corneal wound healing are described by the present work.
Insights
This study maps corneal cell types during wound healing using single-cell RNA sequencing. It reveals distinct inflammatory and recovery phases, crucial for understanding ocular disease treatment and corneal repair.
Area of Science:
- Ophthalmology
- Cell Biology
- Genomics
Background:
- A detailed cell map is vital for treating ocular diseases and promoting corneal healing.
- Understanding the cellular dynamics of corneal wound repair is essential.
Purpose of the Study:
- To analyze corneal wound healing using single-cell RNA sequencing (scRNA-seq) in mice after laser injury.
- To identify and characterize diverse cell populations and their dynamics during the repair process.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) was performed on mouse corneas post-infrared laser injury.
- Analysis of 12,956 cells identified 28 distinct cell clusters, including epithelial, stromal, immune, and neural cells.
Main Results:
- Three new epithelial subpopulations emerged by day 3 post-injury, with increased immune cells (neutrophils, macrophages).
- Myofibroblasts and stromal stem cells increased, while differentiated epithelial and stromal cells, endothelial, and Schwann cells decreased.
- By day 21, immune cells and myofibroblasts decreased, with partial recovery of endothelial and Schwann cells, and increased differentiated epithelial, stromal cells, and stromal stem cells.
Conclusions:
- Corneal repair involves two phases: early inflammation/epithelial remodeling and late stromal restoration/regeneration.
- The study describes dynamic cellular interactions critical for effective corneal wound healing.
