title: GSM7866244: PID 0; Mus musculus; RNA-Seq
library_construction_protocol: Spinal cords were harvested from sham or SNI mice at PID 3, PID 7 and PID 14 in ice-cold PBS solution. Cells in the ipsilateral L4-L6 spinal cord dorsal horns were isolated, homogenized using loose and tight pestles in 1 mL ice-cold Ca2+/Mg2+-free HBSS for 15-20 times by simultaneously rotation. The homogenate was passed through a pre-wet 70 μm mesh cell strainer (BD Biosciences, USA) and spun down at 300 g for 5 min at 4 °C. The cell pellet was resuspended in 40% Percoll solution (GE Healthcare #GE17-0891-01) and centrifuged at 500 g for 15 min at 4 °C. Cells in pellet resuspended and incubated in ice-cold FACS buffer and then incubated in ice-cold FACS buffer containing Cd11b-PE (BioLegend, #101208) at a final dilution of 1:300 for 30 min. The cells were then washed with 1 mL of ice-cold FACS buffer. After centrifuging at 800 g for 5 min, the pellet was resuspended in ice-cold FACS buffer. Living cells exhibiting high fluorescence levels of Cd11b were sorted by FACS using a 70 μm chip on the FACSAriaTM Fusion Platform (BD Biosciences, Carlsbad, CA, USA) with Purity mode. Single-cell RNA-Seq libraries was prepared using Chromium Single Cell 3' Library and Gel Bead Kit v3.0 (10X Genomics #PN-1000075), Chromium Single Cell 3′ Library Construction Kit v3.0 (10X Genomics #PN-1000078), Chromium Chip B Single Cell kit (10X Genomics #PN-1000154), Chromium i7 Multiplex Kit (10X Genomics #PN-120262) and chromium controller, following manufacturer's instruction. The samples were sequenced on an DNBSEQ-T7 sequencer.