GSM7299725: AnaVI (P32) skin mouse 2, scRNA-seq; Mus musculus; RNA-Seq
To isolate cells from mouse back skin, samples were dissected and digested in 0.25% collagenase (Sigma) in HBSS for 35 min before 0.25% trypsin-EDTA for an additional 15 min. Single cell suspensions were obtained by scraping the epidermal side gently using a blunt scalpel. After washing with FACS buffer (5% FBS in PBS), cells were filtered through 70- and 45-μm strainers, pelleted, washed with FACS buffer, and incubated with published antibodies for 20 min on ice. To resuspend autologous hair follicle stem cells (HFSCs), 3D cultures were washed with sterile PBS, dissociated into single cell suspensions using TrpLE-Express (Gibco) for 15min at 37°C. After filtering, cells were pelleting, washed with Cell Staining Buffer, and incubated with TotalSeq-B hashtag antibodies for 20 min on ice. Libraries were constructed according to the manufacter's instructions (3' Gene Expression V3, 10x Genomics).