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This approach allowed us to identify the distinct clusters of LSCs and progenitors with exclusively expressed markers, and might apply for translational research on regenerating a normal corneal epithelium and restoring vision. Overall design: We isolated limbal basal cells from two young donors, constructed scRNA-seq libraries, generated RNA-sequencing data using the 10x Genomics platform, and then finally obtained the transcriptome of about 18,000 individual single-cells using Cell ranger (https://10xgenomics.com). 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This approach allowed us to identify the distinct clusters of LSCs and progenitors with exclusively expressed markers, and might apply for translational research on regenerating a normal corneal epithelium and restoring vision. Overall design: We isolated limbal basal cells from two young donors, constructed scRNA-seq libraries, generated RNA-sequencing data using the 10x Genomics platform, and then finally obtained the transcriptome of about 18,000 individual single-cells using Cell ranger (https://10xgenomics.com). 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This approach allowed us to identify the distinct clusters of LSCs and progenitors with exclusively expressed markers, and might apply for translational research on regenerating a normal corneal epithelium and restoring vision. Overall design: We isolated limbal basal cells from two young donors, constructed scRNA-seq libraries, generated RNA-sequencing data using the 10x Genomics platform, and then finally obtained the transcriptome of about 18,000 individual single-cells using Cell ranger (https://10xgenomics.com). 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This approach allowed us to identify the distinct clusters of LSCs and progenitors with exclusively expressed markers, and might apply for translational research on regenerating a normal corneal epithelium and restoring vision. Overall design: We isolated limbal basal cells from two young donors, constructed scRNA-seq libraries, generated RNA-sequencing data using the 10x Genomics platform, and then finally obtained the transcriptome of about 18,000 individual single-cells using Cell ranger (https://10xgenomics.com). 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We identified 11 unique clusters of cells assigned to a putative cell type based on published known biomarkers for differentiation, proliferation and putative epithelial stem cells. As a results, we found 2 Terminally Differentiated Cell (TDC), 3 Post-Mitotic Cell (PMC), 1 Transient Amplifying Cell (TAC), 2 Limbal Progenitor Cell (LPC), Putative Limbal Stem Cell (LSC) and 2 Melanocyte (MC) sub-cell type clusters. Furthermore, we confirmed the trajectory order of LSC differentiation for 9 clusters using Pseudotemporal and Functional PCA analysis. Lastly, we validated each assigned cell subtypes and determined their location in human corneal limbus tissue with 9 markers using RNAscope We were able to reveal the heterogeneity of corneal limbal basal epithelium by defining novel dynamic trajectories for cell types in a pseudotemporal manner. This approach allowed us to identify the distinct clusters of LSCs and progenitors with exclusively expressed markers, and might apply for translational research on regenerating a normal corneal epithelium and restoring vision. Overall design: We isolated limbal basal cells from two young donors, constructed scRNA-seq libraries, generated RNA-sequencing data using the 10x Genomics platform, and then finally obtained the transcriptome of about 18,000 individual single-cells using Cell ranger (https://10xgenomics.com). Next, we performed quality control, filtering and data integration and further downstream analysis such as dimensional reduction, unsupervised clustering, and Differentially expressed gene selection, trajectory analysis and visualization.", "id": "SRP269263" }, "id": "SRX8635113" }, "annotation": { "rawDataCondition": { "anatEntity": { "id": "UBERON:0006761", "name": "corneo-scleral junction" }, "anatEntityAuthorAnnotation": "corneal limbal epithelial basal layer", "cellType": { "id": "CL:0000034", "name": "stem cell" }, "cellTypeAuthorAnnotation": "LSC (Limbal Stem Cell)", "devStage": { "id": "HsapDv:0000237", "name": "third decade stage (human)" }, "devStageAuthorAnnotation": "young donors (adut in HCA)", "sex": "NA", "strain": "NA", "time": "NA", "timeUnit": "NA", "species": { "id": 9606, "name": "human", "genus": "Homo", "speciesName": "sapiens", "preferredDisplayOrder": 1 } }, "physiologicalStatus": "NA" }, "pipelineSummary": { "meanRefIntergenicDistribution": 13.308588, "sdRefIntergenicDistribution": 1.454792, "pValueThreshold": 0.0500, "allUMIsCount": 0, "mappedUMIsCount": 85203, "distinctRankCount": 309, "maxRank": 21462.00 }, "barcode": "" } ], "downloadUrls": [ { "text": "scRNA-Seq droplet-based processed expression values per cell population in Homo sapiens", "href": "https://www.bgee.org/ftp/current/download/processed_expr_values/droplet_based/Homo_sapiens/Homo_sapiens_Droplet-Based_SC_RNA-Seq_read_counts_CPM_SRP269263.tsv.gz", "title": "Homo_sapiens_Droplet-Based_SC_RNA-Seq_read_counts_CPM_SRP269263.tsv.gz" }, { "text": "scRNA-Seq droplet-based H5AD data per cell in Homo sapiens", "href": "https://www.bgee.org/ftp/current/download/h5ad/droplet_based/Homo_sapiens/Homo_sapiens_SRP269263_droplet_based.h5ad", "title": "Homo_sapiens_SRP269263_droplet_based.h5ad" } ] } }