Single-cell RNA-sequencing from stroma in a mouse submandibular salivary gland following 14-day ligation or mock surgery

The submandibular salivary gland stroma makes up only a small portion of the total salivary gland and the stromal response to salivary gland injury has been understudied. We used single-cell RNA-sequencing (scRNAseq) to analyze which cell types are present in ductal ligated and mock surgery salivary glands, how the cell type abundance is altered during injury, and how the transcriptome of those cells is being altered. This will allow us to examine which cell types are important contributors to recovery from salivary gland ductal ligation injury. Submandibular salivary glands were dissociated into single cells followed by two rounds of magnetic activated cell sorting (MACs) to deplete for epithelial cells expressing EpCAM and red blood cells expressing Ter119. The samples then went through one round of dead cell removal using Miltenyi dead cell removal kit.