GSM6481346: In vitro Activated CD8 T Cells; Mus musculus; RNA-Seq
For the Activated sample, CD8+ T cells were collected after 24h in vitro stimulation and subject to dead-cell removal using a MACS® Dead Cell Removal Kit. In brief, ~2 x 10^6 cells were pooled from 24 culture wells and resuspended in 100 µL Dead Cell Removal MicroBeads suspension, following the manufacturer's guidelines thereafter. The purified live-cell suspension (~90% viability) was then washed once with fresh culture medium, and resuspended at ~10^6 cells/mL in fresh culture medium supplemented with 10 ng/mL IL-2. Dead cell removal was not necessary for the 144h Effector-polarized (~90% viability) or Memory-polarized (~70% viability) samples, in which case ~10^6 cells/sample were pooled from 6 culture wells each, washed once with fresh culture media, and resuspended at ~10^6 cells/mL in fresh culture media supplemented with either 10 ng/mL IL-2 (Effector-polarized) or 50 ng/mL IL-15 (Memory-polarized). Cell suspensions were kept on ice and immediately processed for single-cell library preparation. Cell suspensions for each sample were converted to barcoded single-cell cDNA libraries using the Chromium Single Cell 3' v2 Library, Gel Bead, Chip, and Multiplex Kit, following the manufacturer's guidelines, and aiming for a total of 10000 cells per library.