Controlled ovarian stimulation and laboratory procedures
pituitary blockage
Controlled ovarian stimulation was performed using recombinant
follicle-stimulating hormone (r-FSH, Gonal-F®; Serono, Geneva,
Switzerland), with pituitary blockage using a gonadotropin-releasing hormone
(GnRH) antagonist, cetrorelix acetate (Cetrotide®; Merck KGaA, Serono,
Geneva, Switzerland).Follicular growth was monitored using transvaginal ultrasound examination
starting on day 4 of gonadotropin administration. When adequate follicular
growth and serum estradiol levels were observed, leuprolide acetate
(Lupron®; TAP Pharmaceuticals, North Chicago, IL, United States) was
administered to trigger the final follicular maturation. The oocytes were
collected 35 hours later through transvaginal ultrasound ovum pickup.The recovered oocytes were assessed to determine their nuclear status, and those
in metaphase II were submitted to intracytoplasmic sperm injection (ICSI)
following routine procedures (After ICSI oocyte/embryos were cultured until day five when one or two
blastocysts were transfer.