human T cells extracted from spleen of PTCy-treated mice_p
NSG mice were engrafted with human PBMC on day 0 to induce xenogeneic GVHD and treated (n=4) or not (n=3)with cyclophosphamide 100 mg/kg on day +3. Mice were sacrificed on day +6 and human CD3+ T cells were isolated from the spleen by immunomagnetic selection and pooled for each group. NSG mice were engrafted on day 0 with human PBMC and sacrificed on day 6 to harverst spleen. Human CD3+ T cells were isolated from mice spleen by immunomagnetic selection. Treated mice received a single Cyclosphophamide (100mg/kg) i.p. injection on day 3 post-transplantation Isolated cells were washed once with PBS + 0.04% BSA. Cell concentration was adjusted to 1,000 total cells/Âµl and 13,300 cells were loaded on Chromium Controller (10X Genomics). Samples were processed for droplet-based-RNA sequencing and libraries were prepared using Chromium Single Cell 3' Reagent Kits V3 according to manufacturer's protocol (10x Genomics). Amplified cDNA quality controls were performed with an Agilent bioanalyzer and final library profile were checked on Qiaxcel (Qiagen).