Flow cytometric deoxyribonucleic acid analysis of granulosa cells aspirated from human ovarian follicles. A new method to distinguish healthy and atretic ovarian follicles.

Granulosa cell aspirates from human ovarian follicles were analyzed by flow cytometry to determine the fraction of cells in the DNA S-phase of the mitotic cell cycle. The aim of the study was to evaluate if the percentage of granulosa cells in S-phase (the S-fraction) could be used to indicate whether a follicle was healthy or atretic. A highly significant relationship was found between the S-fraction and the concentration of estradiol in the follicular fluid (r = 0.6, P less than 0.001). More than 85% of the follicles having an S-fraction of 16% or greater contained intrafollicular levels of estradiol equal to or greater than 200 ng/ml and had a low androstenedione:estradiol ratio. Conversely, 95% or more of the follicles that had an S-fraction of less than 16% contained low estradiol (less than 200 ng/ml) and had a high androstenedione to estradiol ratio. We conclude that flow cytometric DNA measurements on follicular aspirates provide a reliable and rapid method by which to distinguish healthy and atretic ovarian follicles. Since only a small fraction (less than 5%) of an entire granulosa cell population is required for S-phase analysis, the technique allows the majority of cells to be immediately available or other biochemical studies. Moreover, since excision of ovarian tissue is avoided, the technique may be acceptable for studies on women with normal ovarian function but who are undergoing laparotomy or laparoscopy for some reason.