GSM8187587: HD1 HSC; Homo sapiens; RNA-Seq
Total CD34+ cells (gate: size/singlets/ Zombie-negative/CD34-positive cells) or phenotypic HSC/MPPs (gate: size/singlets/ Zombie-negative/CD34-positive/bottom 10-20% CD38-low cells, CD45RA-) were sorted using the BD FACSAria Fusion. After the sort, up to 20.000 CD34+ cells were pooled with up to 10.000 phenotypic HSC/MPPs from different donors. After centrifugation, cells were resuspended in 47 μl PBS + 0.04% BSA for library prep using the Chromium™ Single Cell 3' reagents. Cells were immediately processed for single-cell sequencing using the Chromium™ Single Cell 3' Library & Gel Bead Kit v3 (10x Genomics) following the manufacturer's protocols. Libraries from multiple samples were combined prior sequencing and sequenced on the Illumina NovaSeq 6000 system, aiming for a minimum of 40,000 reads per cell for gene expression and 10,000 reads per cell for cell surface protein.