Natal dental pulp stem cells (Kocaeli University)

The pulp tissue was digested, using collagenase type I, to form single-cell suspensions. The cells were then cultured in MEM-Earle supplemented with fetal bovine serum (FBS; 15%) and 100 IU/ml penicilin and 100
μg/ml streptomycin. Cells from one tooth were seeded into two 25 cm² plastic tissue culture flasks and incubated for 3 days. On the third day, red blood cells and other nonadherent cells were removed and the medium was replaced. Adherent cells were cultured until they reached 70% confluency and were defined as passage
zero (P0) cells.