The cornea is the clear window that lets light into the eye. It is composed of five layers: epithelium, Bowman’s layer, stroma, Descemet’s membrane and endothelium.
More...The cornea is the clear window that lets light into the eye. It is composed of five layers: epithelium, Bowman’s layer, stroma, Descemet’s membrane and endothelium. The maintenance of its structure and transparency are determined by the functions of the different cell types populating each layer. Attempts to regenerate corneal tissue and understand disease conditions requires knowledge of how cell profiles vary across this heterogeneous tissue. We performed a single cell transcriptomic profiling of 19,472 cells isolated from eight healthy donor corneas. Our analysis delineates the heterogeneity of the corneal layers by identifying cell populations and revealing cell states that contribute in preserving corneal homeostasis. We identified that the expression of CAV1, CXCL14, HOMER3 and CPVL were exclusive to the corneal epithelial limbal stem cell niche, CKS2, STMN1 and UBE2C were exclusively expressed in highly proliferative transit amplifying cells, and NNMT was exclusively expressed by stromal keratocytes. Overall, this research provides a basis to improve current primary cell expansion protocols, for future profiling of corneal disease states, to help guide pluripotent stem cells into different corneal lineages, and to understand how engineered substrates affect corneal cells to improve regenerative therapies.
Overall design: In this study, we provide a high-quality single-cell ribonucleic acid sequencing (scRNAseq) dataset from 19,472 corneal cells isolated from four female and four male donors. Six full donor corneas and the limbi of two donor corneas were manually dissected by layer and dissociated into single cells for single-cell RNA sequencing. Prior to loading the cells on the 10x Chromium controller, cells were rehydrated in rehydration buffer. Cells were then counted to assess cell integrity and concentration. Approximately 9,000 cells (comprising 3,000 cells from each of the three layers) from corneas 01, 02, 03 and 04 were separately loaded by layer and cornea. Furthermore, 3,000 cells (1,000 cells from each of the three layers) from corneas 05 and 06, and 3,000 cells from corneas 07 and 08 (from the limbal samples) were separately loaded. The resulting sequencing libraries were prepared following a standard 10x Genomics protocol. After filtering low quality cells, the transcriptome profiles of 19,472 cells were further analyzed
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