GSM8586893: Day 0 Unwounded Skin Male Control 3; Mus musculus; RNA-Seq
Tissues were harvested: 1) Unwounded samples consisted of two 6mm punch biopsies of the healthy dorsal skin. 2) Wounded skin samples were three 3.5mm punch biopsies of the entire parially-healed full-thickness dorsal skin wounds on POD4. 3) Buccal mucosa samples were collected from both side cheeks inside the mouth. All tissues were then digested using the Whole Skin Dissociation Kit (Miltenyi) according to manufacturer's instructions. Tissues were incubated in enzyme mixture for 2.5 hours at 37°C prior to dissociation on the gentleMACS Dissociator with the gentleMACS C-tubes (Miltenyi; 'h_skin_01' cycle one time). Cells were filtered through 70µm and 30µm Pre-Separation Filters (Miltenyi) prior to resuspension in KBM-Gold (Lonza) and counting for scRNA-seq library prep. Unwounded skin and buccal samples were all >90% viable; live cells were sorted from Day 4 Wound Skin samples by FACS prior to scRNA-seq library prep. Up to 50,000 cells were prepared for scRNA-seq with Chromium Single Cell 5' (10X Genomics) according to manufacturer's protocol.