Stem cells from exfoliated deciduous teeth (SHED), are isolated from pulp tissue of the teeth of 5-12-year-old children. In culture, SHED cells acquired a fibroblast cell-like morphology with few cytoplasmic protrusions and expressed specific markers. When cultured in neural differentiation medium, cells expressed neuron-specific markers.
Pulp tissue of normal exfoliated human deciduous incisors was cut into small pieces and digested in a solution of type I collagenase (3 mg/ml) and dispase (4 mg/ml) for 1 hour. Single-cell suspensions were seeded onto culture dishes and cultured in Dulbecco modified Eagle medium (DMEM) supplemented with fetal bovine serum (FBS, 10%), penicillin (100 U/ml), and streptomycin (100 U/ml).