GSM8633800: HSPC OP-HU; Mus musculus; RNA-Seq
BM cells were obtained by crushing or flushing femurs and tibia in staining media composed of PBS containing 2% heat-inactivated FBS. Red blood cells were removed with RBC Lysis buffer (eBioscience™, 00-4300-54). BM cells were furthered enriched by combination of lineage depletion kit (Miltenyi Biotech, 130-090-858) and CD117 MicroBeads (Miltenyi Biotech, 130-091-224). Library was prepared according to GEXSCOPE Single Cell RNA Library Kit (high density). Briefly, Lin- CD117+ enriched HSPCs (90,000 cells) were dispelled into single cell for further single-cell RNA sequencing using high density microwell chip (containing 250,0000 wells/chip for capturing 20,000-50,000 single cells, Singleron Biotech Co., Ltd, Nanjing, China) for each sample. The GEXSCOPE microchip integrates single cell capture, cell lysis, capture of the cellular mRNA and molecular labeling. The barcoded cDNA is then amplified and used for the construction of single cell NGS libraries.