GSM9326684: scRNAseq healthy CD8+ T cells 4; Homo sapiens; RNA-Seq
Cryopreserved human PBMCs were thawed as indicated in the manuscript, counted, and 0.5E6-1E6 cells were transferred to a new tube, washed with PBS and centrifuged, followed by an incubation with 50μL of Live/Dead Aqua, CD14/CD16/CD19 V500, CD3 APC-R700, CD4 AF488 and CD8a BV605 antibodies in FACS Buffer with 10% Brilliant Stain Buffer for 30 minutes at RT. Cells were then washed and resuspended using FACS buffer followed by a straining through a 35um nylon mesh strainer. Cells were sorted for live CD8+ T cells into eppendorf tubes with RPMI + 10% FBS. Cells were then washed two times with RPMI + 10% FBS to remove EDTA. Library prep done with 10X Chromium Next GEM Single Cell 5' Kit v2 and Chromium Single Cell Human TCR Amplification Kit.