2-2 Clinical course of ART
COS with follicle-stimulating hormone (FSH) (gonal-f, follitropin-alpha, EMD serono, Japan) or human menopausal gonadotropin (hMG) (ferring, Ferring Pharma, Japan) were started on the cycle day (CD) 3 and GnRH antagonist ; cetrorelix acetate (cetrotide) from CD 14. The total duration of treatment was 20 days, with a total of 6625 units of FSH + hMG administered (Figure 1).
On the 19th day of stimulation, we decided to retrieve the oocytes (ovum pick-up [OPU]) 3 days later, and stimulation was continued until the 20th day. A final oocyte maturation trigger was performed with a GnRH agonist (leuprolide acetate [Lucrin]; AbbVie, Illinois, USA) (subcutaneous administration, 1 mg), and 35 hours later, the oocytes were retrieved with a transvaginal needle under venous anesthesia. A total of 31 oocytes (26 × meiosis-2 stage [M2], 2 × meiosis-1 stage [M1], 1 × germinal vesicle stage, and 2 × degenerated oocytes) were obtained. The unadjusted sperm volume was 3.2 mL, the motile sperm concentration was 5 M/mL, and the sperm motility was 17.9%, so insemination of the M2 oocytes was split. The fertilization result from intracytoplasmic sperm injection (ICSI) was 21 (12 × 2 pronuclear stage [PN], 1 × 1 PN, 5 × 3 PN, 3 × degeneration [Deg]) and that for conventional IVF was five (3 × 2 PN, 1 × 3 PN, 1 × Deg). Confirmation of fertilization showed 15 × 2 PN embryos and 1 × 1 PN embryo. Therefore, 8 × 2 PN embryos were cryopreserved and the remaining 7 × 2 PN and 1 × 1 PN embryos were cultured for blastocysts. Three blastocysts were formed by day 7 of culture (3AB, 4AA, 4BC by the Gardner classification).