GSM6756508: 16w TMJ disc, scRNAseq; Mus musculus; RNA-Seq
To isolate TMJ disc cells, the discs were freed from the joint space and isolated precisely from the surrounding tissue using fine forceps and scissors under a stereomicroscope. Next, TMJ discs were digested in 4 mg/ml Pronase (10165921001, Roche, Switzerland) for one hour, followed by digesting in 2 mg/ml Collagenase P (11213857001, Roche, Switzerland) in fetal bovine serum (FBS)-free DMEM for 2 hours. After filtering the cell suspension using 70 μm and 40 μm cell filters, the cell suspension was loaded into Chromium microfluidic chips with 3' chemistry and barcoded with a 10x Chromium Controller. RNA from the barcoded cells was subsequently reverse-transcribed, and sequencing libraries were constructed with reagents from a Chromium Single Cell 3' v2 reagent kit according to the manufacturer's instructions. Sequencing was performed with an Illumina system.