GSM8086567: CD3+ T cells, trained, UES-WH s011; Mus musculus; RNA-Seq
The ipsilateral hemisphere of brains from trained mice and untrained mice (controls) were isolated 14 days post stroke. Brain mononuclear cells were isolated from single cell suspensions by Percoll density centrifugation (37% Percoll, 20 min at 2800 x g without brake, 18°C). Cells were washed, resuspended in PBS/2mM EDTA/2% FCS and stained with fluorochrome-labelled antibodies (anti-CD45 PB (30-F11) Biolegend; anti-CD3 PE (145-2C11), BD Biosciences). After washing, single cells were sorted into 384 so-called primer plates (Single Cell Discoveries) using a BD Aria FACS Fusion (BD Biosciences). Single cells were sorted into 384 so-called primer plates (Single Cell Discoveries) using a BD Aria FACS Fusion (BD Biosciences). The primer plates contain mineral oil and 50 nl droplet with reagents. Plates were recovered with an aluminium sealer, centrifuged for 2 min and snap frozen on dry ice. SIngle cell sequencing (SORT-Seq) was performed by SIngle Cell DIscoveries, Netherlands.