[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.


METHODS
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.


CONCLUSIONS
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.
Zhao
Hu
H
Department of Biomedical and Biomaterial Sciences, West China School of Stomatology, Sichuan University, Chengdu 610041, China.
Chen
Zhiqing
Z
Bai
Wei
W
D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D002648:Child; D003782:Dental Pulp / Q000166:cytology*; D005260:Female; D006801:Humans; D008297:Male