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Objective: To correlate apoptosis of cultured human granulosa- lutein cells (GL cells) with the outcome of IVF (embryo fragmentation and pregnancy rate) and to study the effect of insulin and insulin- like growth factor I (IGF- I) on apoptosis. Design: In vitro assays. Setting: University laboratory and private IVF center. Patient(s): Eighty- one women undergoing IVF. Intervention(s): Purified human GL cells from pooled follicles were cultured for 48 hours in serum- free media with or without insulin and IGF- I. Cumulus cells and mural GL cells were evaluated separately. Main Outcome Measure(s): Detection of apoptosis by using caspACE FITC- VAD- FMK, a fluorescent in situ marker for activated caspases; embryo fragmentation; and pregnancy. Result(s): Age younger than 38 years and successful pregnancy were associated with less apoptosis (33.0% ± 17.2% vs. 43.2% ± 18.0% and 30.2% ± 14.0% vs. 40.4% ± 19.5% , respectively). There was a linear correlation between embryo fragmentation and GL cell apoptosis. Insulin- like growth factor I decreased apoptosis in a dose- dependent fashion. A statistically significant effect (17% decrease) was reached at a dose of 10 nM. Insulin (10 nM) caused a small (8% ) decrease in apoptosis, but this effect did not reach statistical significance. Cumulus cells consistently had < 3% apoptosis. Conclusion(s): [1] Apoptosis of cultured GL cells may be associated with IVF outcome and ovarian reserve and [2] IGF- I decreases apoptosis of cultured GL cells.
Objective: To correlate apoptosis of cultured human granulosa-lutein cells (GL cells) with the outcome of IVF (embryo fragmentation and pregnancy rate) and to study the effect of insulin and insulin-like growth factor I (IGF-I) on apoptosis. Patient (s): Eighty-one women undergoing IVF. Intervention (s): Purified human GL cells from pooled follicles were cultured for 48 hours in serum-free media with Main Outcome Measure (s): Detection of apoptosis by using caspACE FITC-VAD-FMK, a fluorescent in situ marker for activated caspases; embryo fragmentation; and pregnancy (s): Age younger than 38 years and successful pregnancy were associated with less apoptosis (33.0% ± 17.2% vs. 43.2% ± 18.0% and 30.2% ± 14.0% vs. 40.4% ± 19.5%, respectively). There was a linear correlation between embryo fragmentation and Insulin-like growth factor I decreased apoptosis in a dose-dependent fashion. A significant effect (17% decrease) was reached at a dose of 10 nM. Insulin (10 nM) caused a small (8%) decrease Conclusion (s): [1] Apoptosis of cultured GL cells may be associated with IVF outcome and ovarian reserve and [2] IGF-I Decreased apoptosis of cultured GL cells.