MGCs were obtained from large follicles at the time of oocyte retrieval from women.
< 40 years of age undergoing IVF treatment due to mechanical problems, endometriosis (confirmed by surgery or ultrasound), pre-implantation genetic disorders, or male factors. Stimulation protocols were used for the induction of follicular growth as previously described [19 (link)]. All patients underwent suppression using a GnRH antagonist protocol (0.25 mg/day, Cetrorelix, Cetrotide; Merck-Serono, Darmstadt, Germany). Ovarian stimulation was performed with a daily subcutaneous dose of recombinant FSH (either Gonal-F; Merck Serono, Darmstadt, Germany,or Puregon Pen,Schering Plough, North Wales PA, USA), which was commenced on the third day of the menstrual cycle and was continued for 5 days. This was followed by a daily dose of human menopausal gonadotrophin (Menogon, Ferring, Switzerland). The initial dose used was dependent upon age, body mass index and prior IVF treatment history. When 3 leading follicles had reached 18 mm in diameter, patients received 250 μg human chorionic gonadotrophin (hCG) (Ovitrelle, Merck-Serono, Darmstadt, Germany).
Oocyte retrieval was scheduled for 36 h after hCG injection and performed by transvaginal ultrasound-guided needle aspiration. FFs were collected in culture tubes containing flushing medium (MediCult).
< 40 years of age undergoing IVF treatment due to mechanical problems, endometriosis (confirmed by surgery or ultrasound), pre-implantation genetic disorders, or male factors. Stimulation protocols were used for the induction of follicular growth as previously described [19 (link)]. All patients underwent suppression using a GnRH antagonist protocol (0.25 mg/day, Cetrorelix, Cetrotide; Merck-Serono, Darmstadt, Germany). Ovarian stimulation was performed with a daily subcutaneous dose of recombinant FSH (either Gonal-F; Merck Serono, Darmstadt, Germany,or Puregon Pen,Schering Plough, North Wales PA, USA), which was commenced on the third day of the menstrual cycle and was continued for 5 days. This was followed by a daily dose of human menopausal gonadotrophin (Menogon, Ferring, Switzerland). The initial dose used was dependent upon age, body mass index and prior IVF treatment history. When 3 leading follicles had reached 18 mm in diameter, patients received 250 μg human chorionic gonadotrophin (hCG) (Ovitrelle, Merck-Serono, Darmstadt, Germany).
Oocyte retrieval was scheduled for 36 h after hCG injection and performed by transvaginal ultrasound-guided needle aspiration. FFs were collected in culture tubes containing flushing medium (MediCult).