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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 [18] criteria before and after preparation.

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A long oocyte preincubation (9–11 hours) prior to ICSI is thought to have bad effects on embryo quality [2], probably due to oocyte ageing.Oocytes are retrieved prior to ovulation in the procedure of IVF or ICSI.According to some reports [13, 14], preovulatory oocytes are not fully mature, even though a first polar body is present. Cytoplasmic maturity is thought to be asynchronous with nuclear maturity in stimulated cycles [15, 16].We considered as “TOP” embryos those seen fertilized at day 1 and were regular 4 to 5 cell embryos at day 2 with less than 20% fragmentation and without any multinuclear blastomeres.The percentage of TOP embryos was defined as the ratio between the number of top embryos and the total number of embryos.

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