Polar body transfer restores the developmental potential of oocytes to blastocyst stage in a case of repeated embryo fragmentation
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)  Apoptosis of cultured GL cells may be associated with IVF outcome and ovarian reserve and  IGF-I decreases apoptosis of cultured GL cells.