GSM6284672: Epidermis_multiome_S2_RNA; Homo sapiens; RNA-Seq
Healthy whole skin biopsies were obtained from two male donors of fair-skin type (55 and 72 y/o) and preserved in MACS Tissue Storage Solution (Miltenyi Biotec). Samples were subsequently cut into small pieces that were further processed using the human Epidermis Dissociation Kit (Miltenyi Biotec) and the Gentle MACS Dissociator (Miltenyi Biotec), following the manufacturer's instructions. Nuclei were isolated from the resulting single-cell suspension using the lysis buffer recipe described in (Wysocka et al., 2001) In brief, epidermal cells were resuspended in Buffer A (10 mM HEPES pH 7.9, 10 mM KCl, 1.5 mM MgCl2, 0.34 M sucrose, 10% glycerol, 1 mM DTT, and 1X protease inhibitor cocktail) containing freshly added Triton X-100 at a final concentration of 0.1% . Cells were resuspended in a 1x106 cells/25 µL of Buffer A ratio and were incubated for 10 minutes on ice. Nuclei were recovered by centrifugation at 1,300 g for 5 minutes at 4ºC and resuspended in 1X Nuclei Buffer (10X Genomics). Approximately 10,000 nuclei per sample were loaded into the Chromium Controller system, and libraries were subsequently prepared using the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Kit (10X Genomics), following the manufacturer's instructions. Libraries were sequenced on a NovaSeq 6000 device (Illumina) using paired-end (28+90bp) settings for 100 cycles for the scRNA-seq, and paired-end (50+50bp) for 100 cycles for the scATAC-seq.