GSM7776247: Hemanoid_d8_IL3 (CD34+); Homo sapiens; RNA-Seq
EB and adherent myeloid-cell-forming-complexes were dissociated with TrypLE Express (#12605028, Thermo Fischer Scientific) for 10-30 minutes at 37°C and filtered through a 70 µm mesh. Subsequently, the single-cell suspension was stained for CD34 expression (#11-0349, 1:50, eBioscience) and DAPI (Sigma) and further enriched by sorting (FACS Aria, BD) for live and CD34+ cells. Library preparation for single cell mRNA-Seq analysis was performed according to the Chromium NextGEM Single Cell 3ʹ Reagent Kits v3.1 User Guide (Manual Part Number CG000204 Rev B; 10x Genomics). A twofold excess of cells was loaded to the 10x controller in the specified volume in order to reach a target number of 5000 cells per sample. Fragment length distribution of generated cDNAs and libraries was monitored using 'Bioanalyzer High Sensitivity DNA Assay' (5067-4626; Agilent Technologies). Quantification of libraries was performed by use of the 'Qubit® dsDNA HS Assay Kit' (Q32854; ThermoFisher Scientific).