Semen DNA fragmentation index, evaluated with both TUNEL and Comet assay, and the ICSI outcome.

BACKGROUND: In intracytoplasmic sperm injection (ICSI), there is always a risk of using spermatozoa with damaged DNA. The purpose of this study was to evaluate the percentage of spermatozoa with DNA fragmentation in processed semen samples used in ICSI cycles and to investigate the relationship between the DNA fragmentation index (DFI) and the ICSI outcome. PATIENTS AND METHODS: Fifty-six couples undergoing ICSI treatment were included. DFI was evaluated, by both terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) and single cell gel electrophoresis (Comet) assays, in the processed semen samples used for ICSI. RESULTS: Of the processed semen samples 17.85% had > or =10% spermatozoa with fragmented DNA. There was no correlation between DFI and the ICSI outcome. DFI assessed by the TUNEL assay was negatively correlated with sperm concentration, progressive motility and sperm morphology. CONCLUSION: A considerable proportion of processed semen samples used for ICSI have a high DFI. However, DFI of the processed semen samples does not seem to be related to the ICSI outcome.

Gonadotropin-releasing hormone antagonists do not influence the secretion of steroid hormones but affect the secretion of vascular endothelial growth factor from human granulosa luteinized cell cultures.

OBJECTIVE: To evaluate the secretion of E(2), P, and vascular endothelial growth factor (VEGF) in human granulosa luteinized cell cultures with the presence of a gonadotropin-releasing hormone (GnRH) agonist or antagonist. DESIGN: In vitro cell culture study. SETTING: Research laboratory of a university hospital. PATIENTS: Granulosa luteinized cells were obtained from 24 patients undergoing ovarian stimulation for IVF treatment. INTERVENTIONS: Granulosa cells were cultured for 48 hours with 1 nM of cetrorelix or leuprorelide. For a further 48 hours, granulosa cells were cultured with or without the combination of cetrorelix plus leuprorelide. MAIN OUTCOMES: At the end of each culturing period, the concentrations of E(2), P, and VEGF were measured in culture supernatants by immunoassays. RESULTS: Estradiol and P concentrations were similar between the culture supernatants from controls and treatment groups. The VEGF concentrations in supernatants from cultures with cetrorelix (2,315.1 +/- 1,565.5 pg/mL) were moderately, but significantly, lower than in controls (2,604.3 +/- 1,907.1 pg/mL) or cultures with leuprorelide (2,558.8 +/- 1,403.1 pg/mL). CONCLUSIONS: The GnRH analogues do not affect steroidogenesis in human granulosa luteinized cell cultures. The GnRH antagonists moderately affect the secretion of VEGF from human granulosa luteinized cells.