GSM9979884: U3013MG, cultured brain-slice PDCX, day 1, dasatinib treatment, tech rep 2; Homo sapiens; Mus musculus; RNA-Seq
The four slices per sample (each slice from different brain of U3013MG PDCX) for each time point were pooled together using cold HBSS buffer containing 1% Pen/Strep, 0.6% glucose, and 25 nM HEPES. The tissues of brain slices were dissociated into single cells by mixing with 1 ml micropipette tip and using tissue dissociation kit, human and mouse (Miltenyi, #130-095-942) according to the manufacturer's protocol. Red blood cells were removed using the Red Blood Cell Lysis Solution (Miltenyi, #130-094-183). The myelin debris was removed using myelin removal kit (Miltenyi, #130-096-733). The single-cell RNA sequencing libraries were prepared according to the manufacturer's guidelines, following the Chromium GEM-X Single Cell 3' Kit v4. (10X Genomics, #PN-1000686). Following cell isolation from PDCX slices, cells were washed and resuspended in 1X PBS and loaded onto Chromium GEM-X Single Cell 3´chip (10X Genomics, #PN-1000690) aiming to recover 20000 cells per sample. The quality of the libraries was assessed using Agilent High Sensitivity D5000 Kit (Agilent Technologies, #5067-55(92,93,94)).