Rfsh gonal f

FF was obtained from women who underwent the standard short protocol of IVF
(using a GnRH antagonist - Cetrotide^®, Merck, Germany).
Controlled ovarian stimulation was performed using recombinant FSH (rFSH -
Gonal-F^®, Merck, Germany) and was monitored with
ultrasound. Ovulation was triggered with a GnRH agonist (aGnRH - Gonapeptyl,
Ferring, Germany) when at least two follicles reached 20 mm. The FF was
collected from the dominant follicles through aspiration between 34 and 36 h
after aGnRH administration, using transvaginal ultrasound guidance. Only clear
FF samples, without blood or flushing medium contamination, were processed. The
selected FF samples were centrifuged at 1200 rpm for 10 to 15 min to remove
cellular debris. The supernatants were stored at −80ºC until
purification.

For the controlled ovarian hyperstimulation (COH) procedure, a combination of gonadotrophin was administered with a long GnRH protocol using one of the three following regimens: 1. rFSH Gonal F (Merck KGaA, Darmstadt, Germany) in three dose groups (30IU, 50IU, and 70IU), 2. GnRH agonist (suprefact) (Sanofi S.A., Paris, France). 3. hCG (Pregnyl; Merck KGaA). The GnRH agonist was administered at a dose of 160 µg beginning in the luteal phase in the middle of the previous menstrual cycle and continued until the day before ovulation (approximately 14 days). After obtaining the E2 hormone level <70 pg/mL on the second day of menstruation, the administration was combined with rFSH at the dose of 30, 50, and 70 IU for the three treatment groups. rFSH was injected on the second day after menstruation at a dose according to the treatment group for 10 days until E2 secretion had peaked. Furthermore, hCG was administered at a dose of 10,000 IU or equivalent to 3200 IU. The luteal phase was determined by measuring serial P4 levels starting on the post ovulation day (Figure 1).