scRNA-seq of murine tendon from sham and injured group

ScRNA-seq was performed on murine single cell suspension collected from sham and injured group. Tendon tissues were collected and dissected into pieces. Then they were digested in a mix of collagenase II (Roche, 3 mg/ml) and dispase II (Sigma-Aldrich, 4 mg/ml), prepared in DMEM for 1 hour at 37 °C. Digestions were subsequently quenched with 10% FBS DMEM and filtered through 40μm sterile strainers. Cells were then washed in PBS with 0.04% BSA, counted and resuspended at a concentration of ~1000 cells/μl. Cell viability was assessed with Trypan blue exclusion on a Countess II (Thermo Fisher Scientific) automated counter and only samples with >85% viability were processed for further sequencing.