Dental pulp stem cells (Dayananda Sagar College of Dental Sciences)

Teeth surfaces were cleaned and cut to reveal the pulp chamber, and pulp tissue was then gently isolated and placed in PBS solution. The tissue was digested in a solution of collagenase (3 mg/ml) and dispase (4 mg/ml) for 1 h. Cells were removed and resuspended in Dulbecco’s PBS (DPBS) and centrifuged at 1,800 rpm for 5 min to pellet the cells. Cells were collected and resuspended in DMEM knockout medium and cultured for several days to promote cell adhesion. The medium was then replaced with complete DMEM knockout medium supplemented with FBS (10%).