GSM4119767: sham blood 1; Mus musculus; RNA-Seq
Single-cell suspensions were prepared from mouse blood and brain. Peripheral blood was obtained from deeply anesthetized mice by cardiac puncture, and 700 μL of blood was diluted in 1300 μL of HBSS and 100 μL of 1% heparin sodium. The diluted blood was fractioned in 2 mL Ficoll solution at 500 g for 15 min. Mononuclear cells in the middle layer were collected. Mice were transcardially perfused with sterile 0.9% NaCl after blood collection. The ipsilesional brain hemisphere was harvested, and cell suspensions were prepared using the Neural Tissue Dissociation Kit and gentleMACS Octo Dissociator with Heaters (Miltenyi Biotec). Suspensions were passed through a 70 μm cell strainer, and fractionated on a 30% and 70% Percoll gradient at 500 g for 30 min. Mononuclear cells at the interface were collected. Isolated mononuclear cells were washed, resuspended at 1×10^6 per mL, and stained with fluorophore-conjugated antibodies and the appropriate isotype controls. Flow cytometry was performed using the FACS Aria I sorter and FACS Diva software (BD Biosciences). Monocytes and macrophages were gated as CD11c–Ly6G–CD11b+CD45high and sorted for bulk RNA-seq. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM4119767