title: GSM8059099: HSPC mid endocytosis, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: CB HSPCs were sorted after thawing (7AAD- CD34+ CD38- CD90+) and cultured for 96 hours. The cells were sorted again for HSPC phenotype (DAPI- CD34+ CD38- CD90+ EPCR+) in 3 fractions based on the intensity of internalized fluorescent dextran (20% lowest, mid, 20% highest). Cells were sorted into PBS 0.04% Ultrapure BSA (Thermofisher Scientific AM2616) using a BD FACS Aria cell sorter. A Chromium single cell instrument (10x genomics) was used for the generation of single-cell gel beads in emulsion. In brief, single-cell suspension of cells in 0.4% BSA-PBS were added to each channel on the 10X chip. Cells were partitioned with Gel Beads into emulsion in the Chromium instrument where cell lysis and barcoded reverse transcription of RNA occurred following amplification. scRNA-seq libraries were prepared by using the Chromium single-cell 3′ library and gel bead kit v3 (10x Genomics).