The more accurate assessment of the age of wild animals is extremely important for understanding its population age structure and vitality, the correctness of creation and implementation of management plans, as well as the assessment of trophy animals. In the case of the roe deer, several age estimation methods are available based on external features, as well as characteristics of the teeth, antlers, epiphyses, and bone sutures. The referral (most reliable) method is the counting of incremental growth lines in tooth cementum, on decalcified tooth sections stained with the Harris method. In this research, the age of 37 roe buck from hilly and lowland habitats was determined. The age estimated by the referral method was compared with five other methods: 1) counting growth lines on the native tooth section, 2) incisor inclination, 3) M2 crown height and pedicle diameter, 4) crown height of all teeth and pedicle diameter, and 5) crown heights of M2 and P3. The average age of bucks estimated by the referral method was 4.7 ± 1.68 years, with a range from three to 11 years. The actual accuracy of other methods compared to the referral one was 2.7% (3rd and 4th method), 11.6% (2nd), 21.6% (1st) and 48.6% (5th). According to results, the method of comparing the heights of M2 and P3 crowns is the most reliable with 48.6% accuracy. Additionally, although the actual accuracy of the 3rd and 4th method is low, the deviations from the referral age are relatively small, making them quite reliable. It was also established that the most accurate age estimation by all methods was achieved for roe bucks up to the age of four years, after which the estimation error increases in a way that with most methods the roe deer are presented as younger than they really are.