We assessed sources of variation in the use of tooth cementum layers to determine age by comparing counts in premolar tooth sections to known ages of 20 sea otters (Enhydra lutris). Three readers examined each sample 3 times, and the 3 readings of each sample were averaged by reader to provide the mean estimated age. The mean (SE) of known age sample was 5.2 years (1.0) and the 3 mean estimated ages were 7.0 (1.0), 5.9 (1.1) and, 4.4 (0.8). The proportion of estimates accurate to within +/- 1 year were 0.25, 0.55, and 0.65 and to within +/- 2 years 0.65, 0.80, and 0.70, by reader. The proportions of samples estimated with >3 years error were 0.20, 0.10, and 0.05. Errors as large as 7, 6, and 5 years were made among readers. In few instances did all readers uniformly provide either accurate (error <1 yr) or inaccurate (error >1 yr) counts. In most cases (0.85), 1 or 2 of the readers provided accurate counts. Coefficients of determination (R2) between known ages and mean estimated ages were 0.81, 0.87, and 0.87, by reader. The results of this study suggest that cementum layers within sea otter premolar teeth likely are deposited annually and can be used for age estimation. However, criteria used in interpreting layers apparently varied by reader, occasionally resulting in large errors, which were not consistent among readers. While large errors were evident for some individual otters, there were no differences between the known and estimated age-class distribution generated by each reader. Until accuracy can be improved, application of this ageing technique should be limited to sample sizes of at least 6-7 individuals within age classes of >/=1 year.