GSM5743970: Normal_Dermis; Homo sapiens; RNA-Seq
Freshly collected human skin biopsies were immediately washed with cold PBS and digested with 1X dispase (Thermo Fisher Scientific) overnight at 4 °C. The epidermis and the dermis were separated from each other using forceps and then digested at 37 °C with 0.25% trypsin/EDTA. Digestion efficiency was monitored every 5 minutes by microscopic examination of cell counts in solution. The epidermal and dermal cell suspensions were passed through 70 µm and then 40 µm cell strainers, precipitated via centrifugation at 300 g centrifugal force for 5 minutes, washed, and suspended in cold 0.4%BSA in keratinocyte serum free media (Thermo Fisher Scientific). Single-cell suspensions were loaded on the 10x Genomics system, and mRNA was converted to cDNA library using 10x 3' scRNAseq kit, v3 and S1 flow cell with reads 28x8x91