The purpose of this investigation was to determine the impact of heritability on third molar agenesis in twins. The study sample consisted of 284 same sex twins (172 monozygotic and 112 dizygotic), whose mean ages were 19.7 ± 4.3 and 18.9 ± 4.8 years, respectively. The monozygotic group consisted of 36.3% males and 63.7% females, while the dizygotic group consisted of 50.1% males and 49.9% females. The zygosity of the twins was established using 15 specific DNA markers. The prevalence of third molar agenesis in monozygotic twins was 19.6%, which was higher than in the dizygotic twins group (15.50%) (p = 0.004). In both groups, third molar agenesis was more frequent in the maxilla than in the mandible (p = 0.000). Agenesis of the maxillary third molars was mostly affected by additive genetic factors (62-63%), with the common environment and the specific environment accounting for up to 25% and 13%, respectively. In contrast, agenesis of the lower third molars was associated with a higher additive genetic determination (81-83%), with the specific environment accounting for 17% to 19%. The study's conclusion is that the formation of the third molars follicle is strongly controlled by additive genetic factors.