GSM8946499: G-CSF mobilized graft and PBMC post-alloHSCT sublib 2; Homo sapiens; RNA-Seq
Peripheral blood stem cell graft samples were collected at the time of transplantation and were cryopreserved on the next day. The PBMC samples of recipients were collected at day +30 and day +100 and were cryopreserved on the same or the following day. Mononuclear cells were isolated using a Ficoll gradient and cryopreserved in DMSO until they were used for single cell analysis. Ten adult (>18y) patients with AML in complete remission who underwent alloHSCT with peripheral blood stem cell grafts. Peripheral blood stem cell graft samples were collected at the time of transplantation and were cryopreserved on the next day. The PBMC samples of recipients were collected at day +30 and day +100 and were cryopreserved on the same or the following day. Mononuclear cells were isolated using a Ficoll gradient and cryopreserved in DMSO until they were used for single cell analysis. All live cells (DAPI⁻) were sorted from frozen PBMC samples. The sorted cells were fixed and permeabilized using Evercode™ Cell Fixation v2 (Parse Biosciences, Cat. ECF2001) following the manufacturer's protocol. Fixed cells were stored at −80°C. All samples were then processed together to prepare single-cell libraries using one Evercode™ WT Mega v2 Kit (Parse Biosciences, Cat. ECW02050), according to the manufacturer's instructions. Fifteen sub-libraries were sequenced on the Illumina NovaSeq 6000 S4 flow cell platform using PE150 chemistry with a sequencing depth of 25,000 reads per cell.