Objective To determine whether measures of ovarian reserve differ between females exposed to cancer therapies in a dose-dependent manner as compared with healthy controls of similar age and late reproductive age. Design Cross-sectional analysis of data from a prospective cohort study. Setting University medical center. Patient(s) Seventy-one cancer survivors aged 15–39 years; 67 healthy, similarly aged unexposed subjects; and 69 regularly menstruating women of late reproductive age (40–52 years). Intervention(s) None. Main Outcome Measure(s) Early follicular-phase hormones (FSH, E2 , inhibin B, antimüllerian hormone AMH) and ovarian ultrasound measurements (ovarian volume and antral follicle counts AFC) were compared using multivariable linear regression. Result(s) In adjusted models, FSH, AMH, and AFC differed between exposed vs. unexposed subjects (FSH 11.12 mIU/mL vs. 7.25 mIU/mL; AMH 0.81 ng/mL vs. 2.85 ng/mL; AFC 14.55 vs. 27.20). In participants with an FSH <10 mIU/mL, survivors had lower levels of AMH and AFC compared with controls. Alkylating agent dose score was associated with increased levels of FSH and decreased levels of AMH. Exposure to pelvic radiation was associated with impairment in FSH, AMH, AFC, and ovarian volume. Antimüllerian hormone was similar in women previously exposed to high-dose cancer therapy and 40–42-year-old controls. Conclusion(s) Measures of ovarian reserve are impaired in a dose-dependent manner among cancer survivors compared with unexposed females of similar age. Reproductive hormone levels in menstruating survivors exposed to high-dose therapy are similar to those in late-reproductive-age women. The predictive value of measures for pregnancy and menopause must be studied. ClinicalTrials.gov identifier NCT01143844.