title: GSM8334156: U3013MG, PDX, replicate 1, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: Upon the experimental endpoint, mouse brains were harvested into cold HBSS buffer containing 1% Pen/Strep, 0.6% glucose, and 25nM HEPES. Then, the brain was sliced using a 1mm coronal section matrix, cut into about 1-2 mm pieces using a surgical blade, and dissociated into single cells using the Tumor Dissociation kit, human (Miltenyi, #130-095-929), used in combination with the Mouse Cell Depletion Kit (Miltenyi, #130-104-694) according to the manufacturer's protocols. Red blood cells were removed using the Red Blood Cell Lysis Solution (Miltenyi, #130-094-183 The generation of single-cell RNA sequencing libraries followed the manufacturer's guidelines, utilizing the Chromium Single Cell 3' Library and Gel Bead Kit v2, v3, and v3.1 (analysis of KO-cells) (10x Genomics, Pleasanton, CA). Cryo-preserved cells underwent washing and re-suspension in 0.1% BSA in PBS just before loading onto a Chromium Single Cell B Chip (10X Genomics) with the aim of capturing 10,000 cells.