Flow Cytometric Analysis of Deoxyribonucleic Acid in Human Granulosa Cells fromIn VitroFertilization Cycles: Relationships to Oocyte Maturity and Fertilizability and to Follicular Fluid Steroids*

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Abstract
We measured the mitotic activity of granulosa cells, sex steroid concentrations in follicular fluids, and the maturity and fertilizability of oocytes from 49 follicles. Flow cytometric measurements of DNA were used to determine the percentage of cells in G0/G1, S, and G2/M phases of the cell cycle. Mitotic index was designated as the percentage of granulosa cells in S+G2/M. The progesterone concentration and the progesterone to estradiol ratio in follicular fluids were inversely correlated to mitotic index (r = -0.506; P < 0.001, and r = -0.320; P < 0.02, respectively). Estradiol and androstenedione levels did not correlate with the mitotic index. The mitotic index was higher in follicles with immature oocytes [25.6 .+-. 2.0% (.+-. SE); n=7] than in follicles with mature oocytes (15.6 .+-. 1.2%; n = 41; P < 0.001). The mitotic index of granulosa cells was lowest in follicles with oocytes that fertilized (15.5 .+-. 1.8%), higher in follicles with oocytes that remained unfertilized (18.5 .+-. 1.3%; P < 0.03), and highest in follicles with oocytes that fertilized abnormality (24.0 .+-. 2.1%; P < 0.02). Differences in maturity or fertilizability of oocytes were not associated with variations in follicular fluid progesterone concentrations. The study supports the concept that mitotic activity is decreased when granulosa cells become luteinized. During early follicular growth it is assumed that estradiol and perhaps androstenedione may be important regulators of cell division. Our findings suggest that progesterone, perhaps acting as an antiestradiol, is more important in controlling granulosa cell division of preovulatory follicles during the late follicular phase.

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