GSM8252902: Whole skin, vehicle, sample 2, scRNAseq; Mus musculus; RNA-Seq
Single-cell suspensions were prepared from 6 mm punch biopsies of full-thickness mouse skin using the Miltenyi Whole Skin Dissociation kit (Miltenyi Biotec). Samples were incubated in enzymes P, D, and A for 3 hours at 37°C. Mechanical dissociation was then performed using the h_skin_01 setting on a Miltenyi gentleMACS dissociator (Miltenyi Biotec) for 20 seconds. Suspension was filtered through a 30 µm filter to remove debris. Cell viability was assessed by staining with ViaStain AOPI Staining Solution (Nexcelom Bioscience), and live/dead cells were counted using a Cellometer Auto 2000 (Nexcelom Bioscience). Volume of each suspension containing 50,000 live cells was centrifuged at 300g at 4°C for 5 minutes and resuspended in 50 mL 0.4% BSA prepared in 1x phosphate-buffered saline. Library construction was performed per manufacturer specifications (Chromium Next GEM Single Cell 3' Gene expression (GEX) library protocol, v3.1). Single-cell RNA-sequencing (3' - v3.1)