AbstractContext: Ovaries surgically removed for fertility preservation served as a source of follicle fluid from human small antral follicles.Objective: The objective of the study was to measure intrafollicular concentrations of anti-Müllerian Hormone (AMH), inhibin-B, progesterone, androstenedione, testosterone, estradiol, and IGF binding protein-4.Setting: The study was conducted at a university hospital.Patients: Patients included 43 women having one ovary removed prior to receiving gonadotoxic treatment due to malignant disease.Interventions: Fluid from 100 follicles (diameter of 3–9 mm) were included.Main Outcome Measures: Intrafollicular concentrations of the measured hormones, their possible intercorrelation, and correlation with age were measured.Results: Concentrations of AMH were unrelated to follicular fluid concentrations of androstenedione and testosterone. There was a significant negative correlation between estradiol, inhibin-B, progesterone, and AMH. In four age groups spanning 11–37 yr, levels of AMH, estradiol, androstenedione, testosterone and inhibin-B remained constant, whereas progesterone showed significant variations. IGF binding protein-4 was unrelated to any other measured hormone.Conclusions: This study was unable to confirm a stimulatory effect of androgens on AMH secretion but did enforce a close intimate correlation between AMH and estradiol expressions in the developing human follicle. The insignificant variation of the AMH concentration with age, even in prepubertal girls, suggests that AMH expression is unrelated to menstrual cycle FSH cyclicity.