title: GSM7809244: VS, T3, scRNAseq; Homo sapiens; RNA-Seq
library_name: GSM7809244
library_construction_protocol: Freshly collected vestibular schwannomas and great auricular nerve were resected and washed twice with 1x PBS (Gibco). Fresh samples were cut into 1 mm3 pieces and enzymatically digested with 10 mL digestion medium containing 1 mg/mL collagenase IV (Gibco) and 1 U/mL DNase I (Sigma). The samples were subsequently incubated at 37°C for 45 min and triturated by pipette for 15 min. The suspended cells were subsequently washed with 20% fetal bovine serum (FBS) in Dulbecco's Modified Eagle Medium (DMEM) sieved through a 50-µm Cell-Strainer nylon mesh (BD) and centrifuged at 300 × g for 5 min. The supernatant was removed, and the pelleted cells were washed twice with MACS buffer (PBS containing 1% FBS, 0.5% EDTA, and 0.05% gentamycin) and resuspended in PBS with 0.04% BSA. The cell suspension of each sample was processed through the Gel Bead Kit V3 (10x Genomics) for library preparation according to the manufacturer's protocol. scRNA-seq libraries were constructed using a Chromium Single Cell 30 V2 or V3 Reagent Kit. The single cell libraries were then pooled and sequenced on Illumina Novaseq 6000 Systems using 150-base-pair paired-end reads. Raw data were converted to FASTQ files using Illumina bcl2fastq (version 2.19.1) and aligned to GRCh38 human reference genome.