ICSI Protocol for Assisted Reproduction

ICSI was conducted as previously described (21 (link)). The women were subjected to controlled ovarian hyperstimulation using a standard long protocol. First, oocyte retrieval was performed 36 h after 250 µg recombinant hCG (Ovitrelle, Merck Serono, Switzerland) administration. Next, oocytes were treated with a G-morpholinepropanesulfonic acid (MOPS) medium (Vitrolife, Kungsbacka, Sweden). The embryos were then denuded with hyaluronidase using G-in vitro fertilization (IVF) PLUS medium (Vitrolife). Embryos were cultured in a G-1 medium (Vitrolife) in a desktop incubator (COOK Medical, Bloomington, IN, USA) for at least 3 days. Progesterone was used for luteal support.