Isolation and Culture of Human Stem Cells

HASCs were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). HUVECs were purchased from the China Infrastructure of Cell Line Resources (Beijing, China). Isolation of HAMSCs was performed following the pancreatin/collagenase digestion method[23 (link)–25 (link)]. HASCs and HAMSCs were cultured in 60-mm plates in αMEM supplemented with 100 U/L penicillin, 100 mg/L streptomycin, and 10% FBS in a humidified atmosphere of 5% CO2 at 37°C. HUVECs were cultured in 60-mm plates in EBM containing 1% FBS and EGM-2 BulletKit in a humidified atmosphere of 5% CO2 at 37°C. Cells from passage 3 were used and culture medium was changed every 3 days. The study protocols were approved by the Ethics Committee of the School of Stomatology, Nanjing Medical University, China (NO.PJ2013-037-001). Informed consent was obtained from all the participants enrolled in this study.