Introduction Given the underrepresentation of female physicians in most specialties and the aim of holistic review in residency applicationsto improve the diversity of matriculating resident physicians in the United States (US) postgraduate medical training programs, we examined the association between holistic review and female resident representation among US postgraduate training programs. Methods We conducted a cross-sectional survey of US postgraduate training programs to inquire about their use of holistic review for resident applications (independent variable). The primary outcome was the percentage of female residents in each program, which was obtained along with other program-level characteristics from the Fellowship and Residency Electronic Interactive Database Access (FREIDA) catalog in April 2023. We limited the analysis to the 10 specialties with the most training spots in 2022, including anesthesiology, emergency medicine, family medicine, internal medicine, neurology, obstetrics and gynecology, orthopedic surgery, pediatrics, psychiatry, and surgery (general). We also examined the interactions between holistic review and specialtyand the percentage of female facultyusing model comparison and simple slopes analyses. Results Of the 3,364 total programs surveyed from the 10 specialties, 222 (6.6%)responded. Responders and nonresponders had similar program-level characteristics, including program type (e.g., university, community), specialty, and reported minimum board examination scores. Of the 222 responders, 179 (80.6%) reported performingholistic review. The percentage of female residents was 49.0% (interquartile range 37.5 to 66.7) in the no holistic review group and 47.8% (35.4 to 65.0) in the holistic review group (median difference 0.9%, 95% confidence interval -6.7 to 8.3). Furthermore, there was no evidence of interaction between holistic review and either the specialty or the percentage of female faculty on the outcome of the percentage of female residents. Conclusions Holistic review of residency applications in this limited sample of US postgraduate training programs was not associated with the percentage of female residents. The role of holistic review in addressing the imbalance of male and female physicians in the healthcare workforce, particularly between specialties, remains unknown.