Purpose To evaluate the transition from a proven slowcooling cryopreservation method to a commercial largevolume vitrification system for human blastocysts. Methods Retrospective analysis of de-identified laboratory and clinical data from January 2012 to present date for all frozen embryo replacement (FET) cycles was undertaken. Cryopreservation of trophectoderm-biopsied or non-biopsied blastocysts utilized during this time period was logged as either slow-cooling, small-volume vitrification, or largevolume vitrification. Blastocyst survival post-warm or postthaw, clinical pregnancy following FET, and implantation rates were identified for each respective cryopreservation method. Results Embryo survival was highest for large-volume vitrification compared to micro-volume vitrification and slowcooling; 187/193 (96.9 %), 27/32 (84.4 %), and 244/272 (89.7 %), respectively. Survival of biopsied and nonbiopsied blastocysts vitrified using the large-volume system was 105/109 (96.3 %) and 82/84 (97.6 %), respectively. Survival for micro-volume biopsied and non-biopsied blastocysts was 16/30 (83.3 %) and 2/2 (100.0 %) respectively. Slow-cooling post-thaw embryo survival was 272/244 (89.7 %). Clinical pregnancy and implantation rates outcomes for non-biopsied embryos were similar between large-volume and slow-cooling cryopreservation methods, 18/39 (46.2 %) clinical pregnancy and 24/82 (29.3 %) implantation/embryo, and 52/116 (44.8 %) clinical pregnancy and 67/244 (27.5 %) implantation/embryo, respectively. Comparing outcomes for biopsied embryos, clinical pregnancy and implantation rates were 39/67 (58.2 %) clinical pregnancy and 50/105 (47.6 %) implantation/embryo and 4/16 (25 %) clinical pregnancy and 6/25 (24.0 %) implantation/embryo, respectively. Conclusions The LifeGlobal large-volume vitrification system proved to be very reliable, simple to learn and implement in the laboratory. Clinically large-volume vitrification was as, or more effective compared to slow-cooling cryopreservation in terms of recovery of viable embryos in this laboratory.

Purpose The purpose of this study was to quantitate changes in seminal volume, sperm count, motility, qualitative forward progression, and total motile sperm cells per ejaculate, across three consecutive ejaculates collected from individuals within 24 h preceding an IVF cycle. Methods Men presenting with oligoasthenozoospermia or asthenozoospemia attempted three ejaculates within 24 h preceding IVF. Ejaculate 1 was produced the afternoon prior to oocyte retrieval, and ejaculates 2 and 3 were produced the morning of oocyte retrieval with 2-3 h between collections. Ejaculates 1 and 2 were extended 1:1 v/v with room temperature rTYBS. Test tubes were placed into a beaker of room temperature water, then placed at 4°C for gradual cooling. Ejaculate 3 was not extended, but pooled with ejaculates 1 and 2 and processed for intracytoplasmic sperm injection (ICSI). Results Out of 109 oocyte retrievals, 28 men were asked to attempt multiple consecutive ejaculations. Among this population, 25/28 (89.3 %) were successful, and 3/28 men (10.7 %) could only produce two ejaculates. Mean volumes for ejaculates 1, 2, and 3 were significantly different from each other (p<0.01); the volume decreased for each ejaculate. Mean sperm counts, motility, qualitative forward progression, and total motile cells per ejaculate for the ejaculates1, 2, and 3 demonstrated the following: ejaculates 2 and 3 were not significantly different, but counts, motility, and total motile sperm were improved over ejaculate 1 (p<0.01). Conclusions Pooling three consecutive ejaculates within 24 h increased the numbers of available motile sperm in this population by 8-fold compared to the first ejaculate alone, facilitating avoidance of sperm cryopreservation and additional centrifugation steps that could affect sperm viability and/or function.

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day 5, 6, and 7 blastocysts in order to determine the efficacy of extended D7 culture in our laboratory. Materials and Methods: Over the span of 3 years, a total of 1915 blastocyst stage embryos (average maternal age of 37 ± 4.1 years) underwent trophectoderm biopsy for genetic testing of ploidy status. Pre-implantation genetic screening (PGS) results were retrospectively compared to day of trophectoderm biopsy. Mature oocytes were fertilized using ICSI and embryos were group cultured in a continuous single culture medium (Irvine Scientific). On Day 3 of embryo development, all multi-cell embryos (≥6 cells) were artificially hatched by laser ablation of the zona pellucida. Hatched embryos were then cultured to the blastocyst stage. Trophectoderm biopsy was dependent on development of a well-defined inner cell mass and trophectoderm. Hatching or completely hatched blastocysts were biopsied on either Day 5, 6 or 7 and subsequently vitrified for future use in a warmed embryo transfer cycle dependent on ploidy status. Biopsied samples were analyzed with comprehensive chromosomal screening involving the use of array comparative genomic hybridization (aCGH) or next generation sequencing (NGS) technology by a reference laboratory. Results: 1915 embryos were biopsied yielding a total of 862 (45%) euploid embryos: 485(56%), 328(38%) and 49(5.7%) from D5, D6, and D7, respectively. The incidence of euploidy was 50%, 40%, and 38% among D5, D6, and D7. Chi-square analysis demonstrated that euploid status is dependent on day of biopsy/blastocyst development with chi square statistic calculated to be 17.5847 and a P value < 0.0001 (Figure 1).