GSM9685926: Lung Monocyte/machrophage cells, CEA10 infected; Mus musculus; RNA-Seq
Mice were intravenously injected with CD45 ab before pulmonary digestion. Lungs were liberase digested (2mg/ml liberase in RPMI) and incubated for 37°C for 30min. Digestion was passed through a 40µm filter to isolate single cell suspention. Cells were washed and resuspended in macrophage buffer (PBS with 0.5% BSA and 2mM EDTA) and incubated with CD11b and magnetic beads. CD11b enrichment was performed according to STEMCELL EasySep protocol. Enriched sample was stained and FACS sorted using FACS Aria II using a 2-step staining protocol: 1 step: CD45+CD64+Dump(negative selected cells)-, 2 step: Viability with CD11b+. Cells were resuspended at 1000cells/µl for 10x genomic sequensing. 10x Genomics Single Cell 3' v3 (CG000315)