Comparative gene expression profile of canine and human invariant Natural Killer T (iNKT) cells at single cell level [scRNA-seq]

We used single cell RNA sequencing (scRNA-seq) to characterize the heterogeneity of peripheral blood-derived canine iNKT cells relative to that of human iNKTs. Canine and human iNKT cells were sorted via Magnetic Activated Cell Sorting (MACS) using a human CD1d tetramer loaded with PBS57. >90% T cells were confirmed to be iNKT by flow cytometry. 10,000 cells were immediately processed for scRNA-seq analysis. The remaining cells were expanded for 3-4 weeks in vitro. At the end of expansion, iNKT purity was confirmed by flow cytometry (>90%), prior to processing for scRNA-seq analysis. Please note that as de-multiplexed raw files are unavailable, mixed sample records were created and individual sample information is provided in the metadata.readme.txt.