The primary culture of human tooth pulp cell]

OBJECTIVE: In this study, we established the optimal condition and duration for the primary culture of the human tooth pulp cell with collagenase digestion method. METHOD: Pulp tissue was removed from healthy young human teeth extracted for orthodontic purposes, and then the pulp was digested by collagenase separately in duration of 30 mins, 1 hr, 2 hrs, 3 hrs. After the digestion, the viability and digestion efficiency were evaluated by microscopy and trypan-blue dying. The cytokeratin and vimentin were immunocytochemically detected to identify the cell phenotype. The tissue explant culture and trypsin digestion were set as controls. RESULTS: After digestion of 1 hr, 2 hrs, or 3 hrs, little tissue residue was left, while there is still much tissue remained after digestion of 30 mins. The viability decreased with the elongation of digestion duration. CONCLUSION: 37 degrees C, continuing stirring and 1 hr of type I collagenase digestion are the optimal conditions for the primary culture of human tooth pulp with digestion method.