[In vitro cultivation of human pulpal fibroblast strains--permanent and deciduous teeth].

Abstract

We succeeded in separating and the cultivating stable monolayer cultures of dental pulp fibroblast strains derived from permanent and deciduous human teeth. Human permanent (n = 67) and deciduous teeth (n = 26) were extracted under acupuncture anaesthesia for the correction of malocclusion. After splitting the teeth, the pulp tissues were carefully removed, placed in tissue culture flasks, and grown in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum (FCS). The human pulpal fibroblasts (HPF) of permanent teeth and deciduous teeth (DHPF) were subcultured. Both the HPF and DHPF appeared to migrate from adherent tissues within 24 to 48 hr after explanation. They proliferated in the pulp explants, and lined up in parallel rows of cells closest to the explant tissue within 7 to 10 days in all of the experimental cases. The outgrowing cells were subcultured at 1.3 x 10(4) cells/cm2 in tissue culture flasks every 4-11 days. They showed vigorous proliferation. The average number of cells in the 6-7 day cultures of HPF were 5.6 x 10(4) cells/cm2 from 3 to 16 passages. It was 4.7 x 10(4) cells/cm2 from 3 to 10 passages with DHPF. However, no difference was observed between HPF and DHPF in the amount of synthesized protein in culture flasks. Furthermore, the growth rate of DHPF was more sensitive than that of HPF to the FCS percentages of the culture media.