Release of sICAM-1 in Oocytes and In Vitro Fertilized Human Embryos

Background: During the last years, several studies have reported the significant relationship between the production of soluble HLA-G molecules (sHLA-G) by 48–72 hours early embryos and an increased implantation rate in IVF protocols. As consequence, the detection of HLA-G modulation was suggested as a marker to identify the best embryos to be transferred. On the opposite, no suitable markers are available for the oocyte selection. Methodology/Principal Findings: The major finding of the present paper is that the release of ICAM-1 might be predictive of oocyte maturation. The results obtained are confirmed using three independent methodologies, such as ELISA, Bio-Plex assay and Western blotting. The sICAM-1 release is very high in immature oocytes, decrease in mature oocytes and become even lower in in vitro fertilized embryos. No significant differences were observed in the levels of sICAM-1 release between immature oocytes with different morphological characteristics. On the contrary, when the mature oocytes were subdivided accordingly to morphological criteria, the mean sICAM-I levels in grade 1 oocytes were significantly decreased when compared to grade 2 and 3 oocytes. Conclusions/Significance: The reduction of the number of fertilized oocytes and transferred embryos represents the main target of assisted reproductive medicine. We propose sICAM-1 as a biochemical marker for oocyte maturation and grading

The predicted probability of live birth in In Vitro Fertilization varies during important stages throughout the treatment: analysis of 114,882 first cycles

Research Question: How much the variability in patients’ response during in vitro fertilization (IVF) may add to the initial predicted prognosis based only on patients’ basal characteristics? Design: Anonymous data were obtained from the Human Fertilization and Embryology Authority (HFEA). Data involving 114,882 stimulated fresh IVF cycles were retrospectively analyzed. Logistic regression was used to develop the models. Results: Prediction of live birth was feasible with moderate accuracy in all of the three models; discrimination of the model based only on basal patients’ characteristics (AUROC 0.61) was markedly improved adding information of number of embryos (AUROC 0.65) and, mostly, number of oocytes (AUROC 0.66). Conclusions: The addition to prediction models of parameters such as the number of embryos obtained and especially the number of oocytes retrieved can statistically significantly improve the overall prediction of live birth probabilities when based on only basal patients’ characteristics. This seems to be particularly true for women after the first IVF cycle. Since ovarian response affects the probability of live birth in IVF, it is highly recommended to add markers of ovarian response to models based on basal characteristics to increase their predictive ability

A novel transnational fresh oocyte donation (TOD) program based on transport of frozen sperm and embryos

STUDY QUESTION: What is the clinical efficacy of an oocyte donation program based on the transportation of frozen semen and embryos between two countries? SUMMARY ANSWER: The transnational oocyte donation program is efficient and reliable and it could provide a first-line strategy to overcome the lack of donors in some countries. WHAT IS KNOWN ALREADY: While there is increasing need for donated oocytes, in many countries the availability of donors is still insufficient to cover the therapeutic demands, and patients are referred abroad for treatment. Since embryo cryopreservation is reliable and efficient, we propose a strategy based on frozen embryos instead of frozen oocytes to satisfy the increasing demand for cross border oocyte donation. STUDY DESIGN, SIZE, DURATION: This is a retrospective cohort study including 630 patients treated from December 2015 to July 2017. PARTICIPANTS/MATERIALS, SETTING, METHODS: Infertile women were treated with elective vitrified-thawed embryo shipping and embryo transfer (ET) between two IVF clinics, one in Spain and one in Italy. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 2617 embryos were created for the 630 patients and the survival rate after warming was 98.5%. After the first ET the live birth rate (LBR) was 30.6%. In 476 patients (75.5%), embryos were transferred at the cleavage stage (Day 2 or 3) and the LBR was 29.2%. Vitrified blastocysts were available for 154 patients (24.5%) and the LBR was 35%. Among patients who did not achieve a pregnancy after the first frozen ET (FET), 92.5% had at least one frozen embryo for successive procedures. 213 patients underwent a second FET. The LBR at the second FET was 30%. The cumulative LBR at the end of the observation period was 39.3%. LIMITATIONS, REASONS FOR CAUTION: The study design was retrospective. A direct comparison with vitrified oocyte donors cycle and subsequent fresh ET would have permitted to compare this strategy versus the current standard based on vitrified gametes. WIDER IMPLICATIONS OF THE FINDINGS: The LBR found in our study is more than acceptable and seems to be higher than what reported with vitrified oocytes. The transnational fresh oocyte donation program may have several advantages over the shipment of vitrified oocytes: similarly to the fresh oocyte donation program it allows for personalized care in oocyte recipient, which is provided by assigning a flexible number of oocytes, and at the same time it maintains the benefit of a frozen ART program permitting scheduling flexibility. The TOD program is efficient and may be proposed as a first-line strategy for distance and inter-countries oocyte donation programs.None

Ultrastructural comparison of in-vitro and in-vivo matured human oocytes

Study question: Is the ultrastructure of the cytoplasm of human in-vitro and in-vivo matured oocytes comparable? Summary answer: Overall, the ultrastructure of human in-vitro matured oocytes is comparable to that of in-vivo matured controls, although following in vitro maturation (IVM) mitochondria-smooth endoplasmic reticulum (M-SER) complexes are partly replaced by mitochondria-vesicles (M-V) aggregates. What is known already: Immature oocytes retrieved from antral follicles of patients undergoing IVM treatment can achieve meiotic maturation in vitro, fertilize and develop into embryos able to implant and give rise to viable pregnancies. However, nothing is known on the ultrastructure of IVM oocytes. Study design, size, duration: The ultrastructure of in-vitro matured oocytes (n = 7) was compared with that of in-vivo matured oocytes (n = 10) by light and transmission electron microscopy (LM, TEM). The study was carried out over a period of 18 months. Participants/materials, setting, methods: Immature cumulus cell-enclosed oocytes, retrieved from mid-sized antral follicles of women requiring IVM treatment, were matured in vitro for 30 h. Mature oocytes were obtained from age-matched women undergoing full ovarian stimulation. In-vitro and in-vivo matured oocytes were fi xed and analysed by LM and TEM. Main results and the role of chance: In-vitro matured oocytes showed general features comparable to in-vivo matured controls. All oocytes had normal ooplasm showing uniform distribution of organelles. M-SER aggregates and M-V complexes were commonly found in in-vivo matured oocytes. Large M-V complexes partially replaced M-SER aggregates in IVM oocytes. Mitochondria appeared morphologically unaffected by IVM. Cortical granules appeared typically stratifi ed in a single, mostly continuous row just beneath the ooplasm in all oocytes. Microvilli were well preserved after IVM. Vacuoles were only occasionally found in all oocytes and, if present, they were frequently associated with lysosomes. The morphological features of the MII spindle and the fi rst polar body of in-vitro matured oocytes were comparable to those shown by control oocytes. Limitations, reason for caution: Although informative and based on a wellestablished methodology, the study should be extended to larger number of oocytes and different maturation conditions. Wider implications of the fi ndings: Ultrastructural analysis offers an objective approach for the comparison of organelle structure and distribution in invitro and in-vivo matured oocytes. The present data confi rm that following IVM the overall oocyte cytoplasmic architecture is well preserved, although subtle differences in comparison to in vivo-matured controls encourage a further refi nement of IVM protocols. Study funding/competing interest(s): Funding by hospital/clinic(s) – Biogenesi, Reproductive Medicine Centre, Monza, Italy. Trial registration number: NA

Production of sHLA-G molecules by “in vitro” matured cumulus-oocyte complex

Oocyte selection with the highest competency represent a major goal in IVF. Several studies have demonstrated that non classical HLA class I HLA-G molecule modulation creates a tolerogenic microenvironment at the feto-maternal interface and it seems to be implicated in embryo implantation. This study investigated if soluble HLA-G molecules production by the cumulus-oocyte complex (COC) could be a marker of oocyte maturation. sHLA-G molecule levels were analyzed using Bio-Rad’s Bio-Plex assay in 152 COC supernatants obtained from 42 women and maturated by an “in vitro maturation procedure”. The presence of sHLA-G molecules was confirmed by Western blot technique. The results demonstrate detectable amounts of sHLA-G molecule ranging from 300 to 800 pg/ml in 14/73 (19%) COCs that generated mature oocytes and the complete absence of detectable sHLA-G antigens in the supernatants of COCs that corresponded to immature oocytes. The detection of sHLA-G molecules in the COC culture supernatants corresponding to matured oocytes is proposed as a possible marker to identify the gametes with a higher functionality. This non-invasive marker could be used in addition to morphological approaches to reduce the number of fertilized oocytes and transferred embryos