Isolation and Cultivation of DPSCs

DPSCs were cultivated and identified according to our previous developed protocol[56 (link)]. Briefly, impacted third molars from healthy individuals (15–25 years old) were collected from the Department of Stomatology, Renmin Hospital of Wuhan University. The study was approved by the Ethics Committee of Renmin Hospital of Wuhan University and informed consent was obtained from all donors. After cleaning and disinfecting the tooth surface with alcohol, the teeth were dissected aseptically, and the pulp tissue was isolated and rinsed in PBS. The dental pulp tissue was cut into small pieces of 1mm³, and digested with type I collagenase and dispase (Sigma, Germany) at 37 °C for 1 h with shaking once every 10 min. After termination of digestion, the cell pellet was resuspended with α-modified Eagle’s medium (α-MEM, Gibco, USA) containing 20% fetal bovine serum (FBS, Gibco, USA), 100 µg/mL streptomycin, and 100 U/mL penicillin (Gibco, USA) and incubated in T-25 culture flasks in a standard culture environment (37 °C, 5% CO₂). The culture medium was changed every 3 days. Cells from passages 3 to 5 were used for experiments.