GSM8086130: Skin0h, scRNAseq; Mus musculus; RNA-Seq
WT mice (n=5, 6~8 weeks) were anesthetized via intramuscular injection of a zolazepam and tiletamine mixture (30 mg/kg, Zoletil 50; Virbac, Carros, France) and xylazine (10 mg/kg, Xylazine Hydrochloride Injection; Bayer, Leverkusen, Germany). The wound was inflicted using a 1.5 mm punch biopsy (Miltex, York, Inc. PA, USA) in the buccal mucosa, with the same size wounds created in the facial skin of the same mouse. At 48h after injury, the oral mucosa and skin wounds were harvested by 3.0 mm punch biopsy. Tissues were minced and digested at 37°C with 0.25% trypsin-EDTA for 10 minutes, and continues to digest at 37°C with RPMI 1640 media containing DNase I (Roche), collagenase type I (Worthington), and dispase (Sigma-Aldrich) for 1 h. Clumps and debris were removed by filtering cells through 70-μm mesh. The single cell suspension was quantified and loaded into the 10X Chromium system, targeting approximately10,000 cells for barcoding in preparation for scRNA-seq using a Single Cell 3′ Library Kit (10X Genomics). Cells were successfully barcoded and their transcriptomes were sequenced using the  Illumina Novaseq System (Illumina, San Diego, CA).