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Study question What is the influence of the ovulation induction strategy on the meiotic status of collected oocytes? Summary answer Non-invasive imaging of meiotic spindles during ICSI showed that ovulation induction with a hCG improved meiotic maturation and subsequent blastocyst formation rate. What is known already Owing to the shorter half-life of GnRH, this strategy has gained attention by significantly reducing risk of ovarian hyperstimulation syndrome. Reduced exposure to LH often requires luteal support and has been associated with a lower live birth rate, a lower ongoing pregnancy rate (pregnancy beyond 12 weeks) and a higher rate of early miscarriage (less than 12 weeks) in fresh autologous cycles, nevertheless. Mid-cycle LH surge triggers molecular activity in follicle cells initiating resumption of meiosis I for competition of the first meiotic division and formation of a MII oocyte inferring competence for subsequent fertilization and further development. Study design, size, duration A retrospective observational study was carried in a cohort of 25 - > 36 years old women including 85 patients. Analysis was carried in two groups: 1: GnRHa trigger and 2: hCG trigger and later stratified for patient age. Participants/materials, setting, methods Women between 25 and > 36 years old, non PCOS, with an AMH > 1.2 ng/ml and stimulated under a short scheme GnRH-antagonist for downregulation, FSH and ovulation induction with GnRHa or hCG. Oocytes allocated to (intracytoplasmic sperm injection) ICSI treatment were cultured for 3-4 hours striped from cumulus cells 2 hours after retrieval and placed in G-MOPS PLUS in aglass bottom dish and with an Octax PolarAide system. Main results and the role of chance A spindle was present in 39.3% (218/554) of all morphologically mature (MII) oocytes alone. Group 1 (GnRHa trigger) and group 2 (hCG trigger) had 56% (309/554) and 44% (245/554) of oocytes at MII stage from which 38% (120/309) and 39% (96/245) showed spindles. A better meiotic maturation stage (scores 3, 4) at ICSI was found when ovulation induction was performed with hCG (group 2) (81% (98/120) versus 88% (85/96), p = 0,0031), nevertheless. Fertilization rate was no different between groups (60% and 58%, respectively), while significantly more blastocysts (at culture day 4 and 5) were generated from group 2 MII oocytes presenting a spindle and a good score (36% (35/98) versus 62% (53/85), p = 0.039). Good blastocyst quality (≥3BB) did not differ between groups. When stratifying for age, women between 25 – 30 presented no differences in presence or meiotic maturation (score 2, 3) in MII oocytes, while fertilization (42% versus 77%, p = 0.022) and blastocyst formation (32% versus 45%, p = 0.012) were better in group 2. Women between > 36 years old presented no differences in presence of meiotic maturation, however, fertilization (40% versus 54%, p = 0.042) and blastocyst formation (25% versus 57%, p = 0.007) were best in group 2. Limitations, reasons for caution Thorough multivariable analysis in a larger patient cohort is required to endorse these results, nevertheless. Wider implications of the findings Prediction of ovulation induction efficacy in different patient cohorts could help fine tuning cycle management. Trial registration number not applicable

Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory.

The sperm-hyaluronan binding assay (HBA) is a diagnostic kit for assessing sperm maturity, function and fertility. The aim of this prospective cohort pilot study was to evaluate the relationship between HBA and WHO sperm parameters (motility, concentration and detailed morphology) and possible influence of sperm processing on hyaluronic acid binding. A cohort of 68 patients undergoing a first combo in vitro fertilisation/intracytoplasmic sperm injection treatment after failure of three or more intrauterine insemination cycles were included in the study. Outcome measures studied were fertilisation rate, embryo quality, ongoing pregnancy rate and cumulative pregnancy rate. HBA outcome improved after sperm preparation and culture, but was not correlated to detailed sperm morphology, concentration or motility. HBA did not provide additional information for identifying patients with poor or absent fertilisation, although the latter had more immature sperm cells and cells with cytoplasmic retention present in their semen. HBA outcome in the neat sample was significantly correlated with embryo quality, with miscarriage rates and ongoing pregnancy rates in the fresh cycles, but not with the cumulative ongoing pregnancy rate. No threshold value for HBA and outcome in combo IVF/ICSI treatment could be established. The clinical value for HBA in addition to routine semen analysis for this patient population seems limited.