GSM8287161: Pig dorsal skin, 8 weeks, wounded and treated with Verteporfin; Sus scrofa; RNA-Seq
Skin wounds from porcine skin and xenografted human skin at POD 14 were harvested and mechanically digested with sharp dissecting scissors (n = 6 per condition). Tissue was then added to an enzymatic digestion solution consisting of Collagenase II (ThermoFisher, Cat:17101015) and IV (ThermoFisher, Cat:17104019) in DMEM-F12 (GIBCOTM, Fischer Scientific, Hampton, NH). Samples were digested on 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 and then filtered a second time using 40 μm cell strainers. scRNA-seq was then performed at the Stanford Genomics Facility (SGF) using the 10x Genomics platform. The relevant protocols were followed per manufacturer's guidelines. Final libraries were analyzed on an Agilent Bioanalyzer High Sensitivity DNA chip for qualitative control purposes. Libraries were sequenced on a HiSeq 4000 Illumina platform targeting 50,000 reads per cell. Transcriptomic samples were prepared from pooled single-cell suspensions using the Chromium Single Cell 3ʹ GEM, Library & Gel Bead Kit v3, Chromium Single Cell B Chip Kit, and Chromium i7 Multiplex Kit (10X Genomics, cat# 1000075, 1000073, and 120262).