GSM8566404: Degos_1_PBMC_TCR; Homo sapiens; OTHER
Skin biopsy from Degos disease patients and healthy volunteers were washed with PBS and digested enzymatically using whole skin dissociation kit (Miltenyi Biotec).  Following mechanical dissociation and filteration, cells were resuspended in 500 μL of complete RPMI-1640 with 10% FBS before sorting. Skin live cells were enriched by fluorescence-activated cell sorting (FACS) using Sytox green. Peripheral blood mononuclear cells (PBMCs) from blood were isolated according to desity centrifugation method. PBMCs and CSF samples were subjected to ACK lysis treatment to remove RBCs. Finally, single cell suspensions prepared from skin, PBMCs and CSF were counted and and cell concentration was adjusted in the range of 700-1200 cells/ul with the viability of > 70% before being used for single-cell capturing. cDNA was generated using 5' 10X kit and subsequenctly used for gene expression library and single cell TCR library preparation. Single cell TCR library construction was performed in accordance with 10X Genomics protocol (10X Chromium 5' v2 or 5'-HT kit).