Olivier Gaspard

Impact of follicular G-CSF quantification on subsequent embryo transfer decisions: a proof of concept study

BACKGROUND Previous experiments have shown that granulocyte colony-stimulating factor (G-CSF), quantified in the follicular fluid (FF) of individual oocytes, correlates with the potential for an ongoing pregnancy of the corresponding fertilized oocytes among selected transferred embryos. Here we present a proof of concept study aimed at evaluating the impact of including FF G-CSF quantification in the embryo transfer decisions. METHODS FF G-CSF was quantified with the Luminex XMap technology in 523 individual FF samples corresponding to 116 fresh transferred embryos, 275 frozen embryos and 131 destroyed embryos from 78 patients undergoing ICSI. RESULTS Follicular G-CSF was highly predictive of subsequent implantation. The receiving operator characteristics curve methodology showed its higher discriminatory power to predict ongoing pregnancy in multivariate logistic regression analysis for FF G-CSF compared with embryo morphology [0.77 (0.69–0.83), P < 0.001 versus 0.66 (0.58–0.73), P = 0.01)]. Embryos were classified by their FF G-CSF concentration: Class I over 30 pg/ml (a highest positive predictive value for implantation), Class II from 30 to 18.4 pg/ml and Class III <18.4 pg/ml (a highest negative predictive value). Embryos derived from Class I follicles had a significantly higher implantation rate (IR) than those from Class II and III follicles (36 versus 16.6 and 6%, P < 0.001). Embryos derived from Class I follicles with an optimal morphology reached an IR of 54%. Frozen-thawed embryos transfer derived from Class I follicles had an IR of 37% significantly higher than those from Class II and III follicles, respectively, of 8 and 5% (P < 0.001). Thirty-five per cent of the frozen embryos but also 10% of the destroyed embryos were derived from G-CSF Class I follicles. Non-optimal embryos appear to have been transferred in 28% (22/78) of the women, and their pregnancy rate was significantly lower than that of women who received at least one optimal embryo (18 versus 36%, P = 0.04). CONCLUSIONS Monitoring FF G-CSF for the selection of embryos with a better potential for pregnancy might improve the effectiveness of IVF by reducing the time and cost required for obtaining a pregnancy.

The actors of human implantation: gametes, embryo and endometrium

The success of pregnancy depends on a receptive endometrium, a normal blastocyst, a synchronized cross-talk at the maternal–fetal interface at the time of implantation, and finally a successful placentation and remodeling of uterine vasculature. In routine, less than 5% of oocytes collected in in vitro fertilization (IVF) cycles and only 20 to 25% of embryos transferred lead to a birth. Implantation and placentation processes remain the black box of fertility, involving following steps: fertilization, endometrial receptivity, embryo implantation (apposition-adhesion-invasion), trophoblastic differentiation and invasion (Cartwright et al., 2010).

CERTOLIZUMAB PEGOL DID NOT RESULT IN A DECREASE IN SEMEN QUALITY IN HEALTHY VOLUNTEERS: RESULTS FROM A PHASE 1 STUDY

Background Effective anti-TNF treatment in large doses may affect sperm count and reduce fertility.1,2,3 More controlled clinical studies to investigate effects of anti-TNF agents on semen quality are required. Objectives To examine the effect of a single dose of certolizumab pegol (CZP) on the semen quality of healthy subjects, to identify the intra- and inter-subject variability in semen quality variables at baseline, and to investigate the safety and tolerability of CZP treatment. Methods 20 healthy males were included in this phase 1, single-centre, double-blind, placebo (PBO)-controlled study (NCT01091220). Subjects were randomized in a 3:1 ratio to receive CZP 400mg (2x200mg subcutaneous injections) or PBO. A screening period (up to 3 weeks) was followed by a 14 week follow up period after the day of dosing. Primary variables for assessment of semen included total motility and morphology (% of normal ovoid forms by Tygerberg criteria4). Treatment effects were assessed using a linear repeated measures model. A mixed effect model (visit - fixed effect; subject - random effect) was applied to the screening data to assess intra- and inter-subject variability. Safety of CZP was evaluated by the number of adverse events (AEs) and routine biochemistry and haemotology laboratory analyses. Results Highest intra-subject variability was found in non progressive motility, spermatozoa count and concentration (66.2%, 43.9% and 36.2%, respectively). Highest inter-subject variability was detected in spermatozoa count, concentration, semen volume and morphology (61.7%, 52.2%, 45.4% and 36.3%, respectively). CZP treatment was found to have no effect on the semen quality variables assessed vs PBO (Table). The changes in semen quality variables during the treatment period were generally within the range of intra- and inter-subject variability seen during screening. The reporting of AEs was broadly similar between the two arms. No AEs were regarded as serious or led to discontinuation from the study. Table 1. Summary of treatment effect on semen quality variables (FAS) Semen quality variable Point estimate (SE) 90% CI Total motility (%) -1.3 (4.3) -8.5; 5.9 Progressive motility (%) -2.1 (4.4) -9.5; 5.3 Nonprogressive motility (%) 0.8 (1.1) -0.9; 2.6 Morphology (%): normal ovoid forms -2.1 (1.5) -4.7; 0.4 Vitality (%) 0.5 (3.4) -5.2; 6.1 Semen volume (mL) -0.2 (0.4) -0.9; 0.5 Spermatozoa count (106) -26.4 (40.8) -94.4; 41.5 Concentration (106/mL) -3.9 (15.1) -29.0; 21.3 CI = Confidence Interval; FAS = Full Analysis Set; SE = Standard Error. Point estimate = difference between treatment and PBO least squares means at post-baseline visits. Conclusions Considerable intra- and inter-subject semen quality variability exists in healthy male subjects. A single 400mg CZP dose was well tolerated and did not produce any treatment effects on semen quality in healthy males compared to PBO. References Pentikainen et al. J Clin Endocrinol Metab. 2001;86(9):4480-82. Suominen et al.Eur J Endocrinol. 2004;151(5):629-40 2004. Mahadevan et al. Inflamm Bowel Dis. 2005;11(4):395-9. World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. Geneva: World Health Organization Press; 2010. Disclosure of Interest S. Perrier d’Hauterive Grant/Research support from: UCB Pharma, S. Kesseler Grant/Research support from: UCB Pharma, P. Ruggeri Grant/Research support from: UCB Pharma, M. Timmermans Grant/Research support from: UCB Pharma, O. Gaspard Grant/Research support from: UCB Pharma, T. Kumke Employee of: UCB Pharma, G. Parker Employee of: UCB Pharma

Comparison between paraffin and mineral oil covering on early human embryo culture: a prospective randomized study

ABSTRACT The oil overlay in microdrop culture systems prevents medium evaporation, helps to maintain appropriate pH and osmotic conditions and protects from microbial contamination. In the present study, we prospectively compared covering by Ovoil™, a paraffin oil, and LiteOil®, a mineral oil, on the in vitro development of human embryos and their suitability for transfer/freezing at day 3 and live birth rate. One hundred and one patients undergoing in vitro fertilization (IVF) treatment by intracytoplasmic sperm injection (ICSI) were enrolled in our study. After ICSI, 1237 oocytes were 1:1 randomly allocated into 2 groups according to the type of overlaying oil: Ovoil™ (616 oocytes) or LiteOil® (621 oocytes). Fertilization rate was assessed around 18 hours post-insemination (hpi) and embryos were checked for early cleavage at 25 hpi. Embryo morphology was recorded on days 2 and 3. A total of 437 (Ovoil™) and 438 day 3 embryos (LiteOil®) were analyzed. There were no differences between the two groups in terms of fertilization rate and occurrence of early cleavage. The proportion of top quality embryos (41.7% vs. 41.2%) and the final utilization rates (92.2% vs. 92.0%) were similar in Ovoil and LiteOil groups, respectively, at day 3. Live birth rate per transfer was essentially the same with Ovoil™ overlay (26.9%) when compared to LiteOil® (26.2%). Live birth rate in patients who simultaneously received embryos from both overlay types was 17.2%. Despite the different characteristics of these two oils regarding hydrocarbon saturation, packing and temperature storage, Ovoil™ and LiteOil® can be used in parallel in the same IVF protocol. Abbreviations: ART: assisted reproductive technologies; hpi: hours post-insemination; hSA: human serum albumin; HTF: human tubal fluid; ICSI: intracytoplasmic sperm injection; IVF: in vitro fertilization; MII: metaphase II; MEA: mouse embryo assay; RT: room temperature.

Sperm Vacuoles: Origin and Implications

Not before the introduction of the Nomarski differential interference contrast optics (DIC), we became aware of the presence of so-called vacuoles on the surface of the sperm head. Before the implementation of motile-sperm organelle-morphology examination (MSOME), no manuscript at all mentioned that spermatozoa free of vacuole-like structures should be selected for injection.