GSM8747080: OM_POD2 scRNA-seq; Mus musculus; RNA-Seq
Mouse oral mucosa and facial wounds were harvested at listed timepoints. Wounds were mechanically digested with sharp dissecting scissors. Tissue was then added to an enzymatic digestion consisting of Collagenase II (ThermoFisher, Cat:17101015) and IV (ThermoFisher, Cat:17104019) in DMEM-F12 (GIBCOTM, Fischer Scientific, Hampton, NH). Samples were added to an orbital shaker at 150 rpm for 90 minutes at 37ºC. FACS buffer was added to quench the digest and samples were filtered through 70 µm cell strainers. Samples were then centrifuged at 1500 for 5 minutes at 4°C. For samples that were sorted, cells were counted and resuspended in FACS buffer. Primary antibodies were applied, and cells were stained in the dark with gentle agitation for 30 min. Cells were then washed thoroughly in FACS buffer. A BD II FACS Aria machine was used for FACS sorting analysis. For lineage negative (Lin-) FACS analysis, the following primary antibodies were used: Tie2, CD45, CD31, CD324, CD326, Ter119. scRNAseq  Stanford Genomics Facility (SGF) using the 10x Genomics Visium platform. The relevant protocols were followed per manufacturer's guidelines.