Human menopausal gonadotropin

Patients were treated with controlled ovarian stimulation according to their medical history as follows. For gonadotropinreleasing hormone (GnRH) agonist cycles, patients received oral contraceptive pills (1 mg of norethisterone and 0.05 mg of mestranol; Aska Pharmaceutical) on day 14 of the previous cycle, which was continued for 10 days, and GrRH agonist (600 mg/day, Suprecur nasal solution 0.15%; Mochida Pharmaceutical) on day 21 of the previous cycle until ovulation induction. On day 3 of the cycle, they received recombinant follicle-stimulating hormone (Gonal F; Merck Serono) ranging from 150 to 300 IU for 4 days followed by human menopausal gonadotropin (Ferring Pharmaceuticals) administration, ranging from 150 to 450 IU, until ovulation induction. For GnRH antagonist cycles, a GnRH antagonist (2.5 mg, ganirelix acetate; MSD) was administered daily after the leading follicles reached 13-14 mm in diameter. Ovulation induction was performed by human chorionic gonadotropin (hCG) administration when at least the leading follicles had reached 18 mm in diameter. Transvaginal follicle aspiration was performed 36 hours after the hCG injection. Insemination was done by a coculture of oocytes with 1.5 Â 10 5 sperm/mL or intracytoplasmic sperm injection 40 hours after hCG injection.

Approximately 2-3 h after retrieval, the cumulus-oocyte complexes (COCs) were denuded of cumulus cells by exposure to HEPES-buffered medium containing 80 IU/ml hyaluronidase (Irvine Scientific, CA, USA), and by pipetting the COCs with a Pasteur pipette. Nuclear maturity of the denuded oocytes was checked. MII oocytes were injected and GV oocytes were cultured in blastocyst medium (G2; Vitrolife Co., Sweden) supplemented with 75 IU/l of human menopausal gonadotropin (Ferring) in a triple gas incubator at 5% O 2 and 6% CO 2 and 89% N 2 . The GV oocytes were assessed for maturity after 24 h of IVM programme. Only 24 h matured oocytes were included in the study and injected using the partner's spermatozoa.