The effects of insulin and parathyroid hormone (PTH) on the proliferation of developing bovine dental pulp in an explant culture system were studied. Dental pulp explants were cultured on siliconized lens paper floating on the serum-free medium for up to 72 h. Ornithine decarboxylase (ODC) activity increased and reached a peak after 24 h. DNA synthesis increased continuously after a lag period of 24 h. Insulin (10 milliunits per ml) stimulated ODC activity 1.3-fold and DNA synthesis 1.5-fold. PTH alone (1 unit per ml) stimulated ODC activity in 1.7-fold, but did not affect DNA synthesis. PTH plus insulin caused greater increases in ODC activity and DNA synthesis in dental pulp explants than insulin alone (ODC, 2.6-fold; DNA, 3.7-fold). These results suggest that insulin and PTH are involved in the regulation of growth of dentinogenically active bovine dental pulp.