title: GSM9604815: isolated living cells from young mouse brain_3; Mus musculus; RNA-Seq
library_construction_protocol: Mice were euthanized and transcardially perfused with PBS, after which brains were rapidly harvested, rinsed in ice-cold PBS, and minced into small fragments. Tissue was enzymatically digested using collagenase, hyaluronidase, and elastase at 37 °C, then filtered through a 100 μm cell strainer to obtain a single-cell suspension. Cells were washed, centrifuged, and enriched for cerebrovascular and neurovascular unit cells using an iodixanol (OptiPrep) density gradient. The enriched cell fraction was collected and resuspended in PBS containing 0.4% BSA for downstream processing. Single-cell suspensions were loaded onto the 10x Genomics Chromium Single Cell 3′ platform for droplet-based capture. Libraries were generated using the Chromium Single Cell 3′ v2 kit following the manufacturer's protocol, pooled based on molar concentration, and sequenced on an Illumina NovaSeq 6000 system. Raw sequencing data were processed using the Cell Ranger pipeline for read alignment, barcode processing, and gene expression quantification