GSM9787147: Healthy human epidermis scRNA-seq donor 2; Homo sapiens; RNA-Seq
Skin samples were obtained from three healthy adult donors during abdominoplasty. From each skin sample, six biopsies were digested with Dispase II overnight at 4 °C to separate the epidermis. Half of the separated epidermis was dissociated using the GentleMACS Human Whole Skin dissociation kit for 2.5 hours at 37 °C according to the manufacturer's instructions. The other half was digested with 0.05% Trypsin-EDTA and DNase I for 7 minutes at 37 °C. Dissociated cell suspensions were pooled, processed using the GentleMACS OctoDissociator, sequentially filtered through 100 µm and 40 µm cell strainers, stained with DAPI for 10 seconds, resuspended in 0.04% BSA in PBS, washed twice, and sorted for viable cells using an AriaFusion high-speed cell sorter. Single-cell gene expression libraries were generated using the 10x Genomics Chromium platform with the Single Cell 3′ v2 chemistry. Immediately after cell sorting, single-cell suspensions were processed for Gel Bead-in-Emulsion generation, cDNA synthesis, and library preparation. GEM generation, library preparation, and RNA sequencing were performed by the Biomedical Sequencing Core Facility of the Center for Molecular Medicine, Vienna, Austria. Biopsy acquisition, cell suspension preparation, and loading on the Chromium instrument were completed within 3.5 hours. Libraries were sequenced on an Illumina HiSeq 3000/4000 instrument.