GSM7497667: Darier Patient 4, lesion skin biopsy; Homo sapiens; RNA-Seq
Slow-frozen skin biopsies were washed with RPMI 1640 medium (Gibco, 61870- 010), supplemented with 10% Gold FBS (PAA Laboratories, A15-151). The skin  biopsies were then minced and transferred in 1mL of Liberase (Roche,  5401119001, final concentration 0.5 mg/mL diluted with PBS without Ca and Mg)  to digest the tissues. The mixture was incubated for 1 hour at 37°C and the tube  was manually inverted every 15 minutes. Afterwards, 100 μl of 0.05% trypsin was added to the mixture and incubated at 37°C for additional 15 minutes. The tissue remnants were removed by using 70μm cell strainer. The cells were washed with  MACS buffer (PBS, 0.5% BSA and 2mM EDTA) and then stained with SYTOX Red  (ThermoFisher, S34859) and Hoechst 33342 (ThermoFisher, H3570) for cell  viability assessment. The stained cells were suspended in MACS buffer and single living cells (SYTOX Red-negative and Hoechst 33342-positive) were sorted by  FACS sorter (BD FACSAris-II SORP) at Flow Cytometry Facility of UNIL.  scRNA-seq processing Single-cell mRNA capture and sequencing were performed immediately by the  Lausanne Genomic Technologies Facility (GTF) of UNIL using the Chromium Next  GEM Single Cell 5' GEM kit (10x Genimics) following the manufacturer's protocol.