Next Generation Sequencing of in vitro mouse T-cell differentiation via VCAM-1. Next Generation Sequencing of in vitro mouse T-cell differentiation via VCAM-1

Sample preparation and single-cell RNA-sequencing of in vitro T-cell differentiation Method: 10X Chromium was used to prepare single-cell cDNA libraries, and Illumina Nextseq was used to 3’ sequence the samples Overall design: Isolated fetal liver cells from decapitated E13.5 CD1 mouse embryos were subjected to TER-119 depletion by EasySep magnetic sorting (STEMCELL Technologies). Next, sorted HSPCs (Sca-1+ cKit+) cultured at 3.1 x10E3 HSPCs/cm2 (corresponding to 1000 cells/well) in DL4 (10 μg/mL) and VCAM-1 (2.32 μg/mL) coated 96-well plates