Objective: This study aimed to establish normal reference ranges for fetal corpus callosum (CC) measured by prenatal ultrasonography (USG) between 16 and 25 weeks of gestation in healthy fetuses. Material and method: A total of 809 fetuses in the second trimester were evaluated by transabdominal USG. All measurements were obtained by a single clinician. Results: Based on the correlation analysis, a significant positive correlation was found between gestational week and CC (r = 0.907, p < 0.001). Conclusion: We anticipate that the reference intervals obtained from healthy fetuses will help clinicians who are interested in neurosonography to detect CC abnormalities at an early stage.