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No more than 7 studies focusing on the influence of ICSI procedure timings on the outcoming results were published [1–8], with discrepancies in the conclusions.Although it has been shown that incubation of oocytes for 2–6 h prior to IVF (In Vitro Fertilization) improves fertilization and pregnancy rates [9–12], there are some conflicting results regarding the timing of ICSI.During all the time of the current study, the ICSI conditions were identical (equipments, media, and culture conditions).Semen samples were collected the day of oocyte retrieval by masturbation.The timing between the end of sperm preparation and the beginning of ICSI was comprised between 30 minutes and 2 hours.Intracytoplasmic sperm injection was performed as described elsewhere .Oocytes are retrieved prior to ovulation in the procedure of IVF or ICSI.
Whether to carry out the oocyte denudation directly after their retrieval or to keep the surrounding cumulus cells during the preincubation is also not clear in the literature.After confirmation of fertilization, each normally fertilized oocyte was transferred into a new ISM1 medium 30 μL droplet. The cleavage rate was defined as the ratio between the number of embryos and the number of mature oocytes.Embryo grading was performed according to the number and the size of the blastomeres (regular or irregular cleavage), the presence or not of multinuclear blastomeres as well as the percentage of anucleate fragments.After liquefaction, semen was prepared by a 90-45 gradient system using Puresperm (JCD, France).Sperm motility and concentration were assessed according to World Health Organization  criteria before and after preparation.
Hence, the fertilizing ability of an oocyte with a mature nucleus is not necessarily at its maximum potential.