GSM8395473: Cd24a+ 10X Genomics scRNAseq; Mus musculus; RNA-Seq
Following euthanasia of mice and tooth extraction, dental pulp was subsequently obtained using a syringe needle to remove epithelial tissue while preserve the mesenchymal part. The collected specimens were then placed into 1.5ml centrifuge tubes, finely minced for 3min, and subjected to a 5min enzymatic dissociation in 0.5ml of 0.25% trypsin-EDTA (ThermoFisher, 25200072). Neutralization was achieved with 1ml of culture medium (DMEM basal medium (Gibco, C11995500BT) supplemented with 10% fetal bovine serum (Lonsera, S711-001)). The medium was further supplemented with 20μl of 5mg/ml DNase I (Roche, 10104159001) and thoroughly mixed. After straining through a 70µm strainer and resuspension in 1x PBS, flow cytometry was employed for the isolation of the targeted cell populations. Cell counting and viability confirmation were performed using the Countess II Automated Cell Counter (Thermo Fisher, AMQAX1000). Samples were then utilized for single-cell RNA sequencing with the 10x Genomics system