To investigate whether behavioral differences exist between female and male wild-type C57 mice in conditioned fear extinction experiments, providing a reference for the equal use of both sexes in fear conditioning and extinction-related studies. Methods: Fear levels, freezing episodes, freezing level per episode, and locomotor activity during conditioned stimulus presentation were measured through auditory-cued fear conditioning, extinction, and retrieval experiments. Behavioral differences between sexes were compared. Results: In fear conditioning and extinction experiments, no significant differences were observed between female and male mice across the four behavioral metrics. During the fear extinction retrieval test, female mice exhibited a higher freezing level per episode upon the first conditioned stimulus presentation, while no significant differences were found in other metrics. Conclusion: Wild-type C57 mice of both sexes show no behavioral differences in fear conditioning and extinction experiments, supporting their equal use in such studies. However, potential sex-based differences in the ability to maintain extinction memory suggest caution when investigating fear memory retention.