Cryopreservation of small numbers of human spermatozoa in a Stripper tip: Report of the first live-birth worldwide.

PURPOSE: The aim is to report the first live-birth following ICSI using spermatozoa previously vitrified in a Stripper Tip. PRINCIPAL RESULTS: A 34-year-old cryptozoospermic man was enrolled in a sperm vitrification program. After failure of conventional freezing technique, spermatozoa were vitrified using two carriers: a commercially-available, Cell Sleeper, and a "home-made" one, Stripper Tip. This man and his 30-year-old wife underwent an ICSI attempt using vitrified-warmed spermatozoa from these devices. All frozen-warmed spermatozoa were quickly recovered. Among the oocytes retrieved, six were injected with sperm from the Cell Sleeper, and seven with sperm from the Stripper tip, leading to 4 embryos in each case. Two embryos, arising from sperm frozen in the Stripper tip, were transferred, resulting in a healthy live-birth. CONCLUSIONS: This is the first successful delivery following the use of spermatozoa frozen in an original device, the Stripper Tip, giving a promising prospect for managing severe male infertilities.

Serum progesterone concentration and live birth rate in frozen-thawed embryo transfers with hormonally prepared endometrium.

RESEARCH QUESTION: Is serum progesterone measurement on the day of embryo transfer associated with outcome of frozen-thawed embryo transfer (FET) in cycles using hormonal replacement therapy (HRT) for endometrium preparation? DESIGN: This single-centre retrospective study assessed the relationship between serum progesterone on embryo transfer day and live birth rates in 227 FET cycles. Endometrial preparation was performed by sequential administration of vaginal oestradiol until endometrial thickness was >7 mm, followed by transdermal oestradiol combined with 600 mg vaginal micronized progesterone. RESULTS: Mean serum embryo transfer day progesterone was 11.4 ng/ml. Serum progesterone <10 ng/ml was observed in 37% of cycles and was associated with significantly lower pregnancy (34% versus 48%, P= 0.04) and live birth rates (17% versus 31%, P= 0.01). Multivariate logistic regression analysis identified serum embryo transfer day progesterone as a significant prognostic factor for live birth rate (odds ratio [OR]: 2.75, 95% confidence interval [CI]: 1.40-5.43]). Receiver operator curve analysis for live birth rates by serum progesterone levels on embryo transfer day gave an area under the curve of 0.62 (95% CI: 0.53-0.72). CONCLUSIONS: The data show that serum progesterone concentration is associated with live birth rate. This outlines the importance of measuring serum progesterone in FET with HRT although progesterone monitoring is not usually performed in routine practice. However, the optimal timing for measurement and further adaptive management in the presence of low values remain to be determined.

Impact of post-warming culture duration on clinical outcomes of vitrified good-quality blastocyst transfers: a prospective randomized study.

OBJECTIVE: To determine whether post-warming culture duration (1 hour vs. 18 hours) influences implantation rates (IRs) of good-quality blastocysts (GQB) in a good-prognosis population. DESIGN: Prospective interventional randomized study. SETTING: University hospital. PATIENT(S): One hundred sixty-two GQB transfers. INTERVENTION(S): Patients' vitrified blastocysts were randomly allocated to group A, warming on the day before transfer (n = 81), or B, warming on the day of transfer (n = 81). MAIN OUTCOME MEASURE(S): IR, live birth rate, reexpansion degree, and quality after warming and immediately before transfer. RESULT(S): Quality of the warmed and transferred blastocysts was similar (respectively, 39.1% and 32.7% top quality [≥B4AA/AB/BA] in group A vs. 41.7 and 42.2% in group B). In group A, 14 of 102 blastocysts (12.2%) appeared to be unsuitable for transfer, versus only 1 of 103 (0.9%) in group B, thus leading to an additional warming. As expected, reexpansion degree just before transfer was higher in group A (0.90 vs. 0.70). Likewise, the proportion of hatched blastocysts before transfer was higher after a longer culture period (38.6% in group A vs. 12.7% in group B). IRs were similar (38.0% in group A vs. 36% in group B), as were live birth rates (35.8% in group A vs. 34.6% in group B). CONCLUSION(S): IRs were not different, whatever the duration of post-warming culture of GQB. Both warming strategies could be applied to good-prognosis patients to optimize the laboratory workflow without any detrimental effect.

Should we consider day-2 and day-3 embryo morphology before day-5 transfer when blastocysts reach a similar good quality?

Clinical outcomes of 291 day-5 blastocyst transfers carried out between January 2012 and March 2016 were retrospectively compared according to their quality at day 2 and 3. Inclusion criteria were female age younger than 37 years; first or second IVF and intracytoplasmic sperm injection cycle; quality of the transferred blastocyst: blastocoele B3 or higher; inner-cell-mass A/B; trophectoderm A/B; and known implantation outcome for each transferred blastocyst. Blastocysts were classified into good-quality and poor-quality embryo groups at day 2 and 3. Implantation (38.7% versus 41.4), clinical pregnancy (40.3% versus 45.9%), miscarriage (22.2% versus 26.7%;) and live birth rates (37.4% versus 38.8%) were comparable in day 2 good and poor-quality embryo groups. No signficiant differences in morphology of transferred blastocysts at day 3 were found. Multivariable analysis highlighted that poor or good embryo quality at day 2 and day 3 were not predictive of the implantation of good-quality blastocysts (at day 2: adjusted odds ratio = 0.82 CI 95% 0.49 to 1.38; at day 3: adjusted odds ratio = 1.39; CI 95% 0.77 to 2.52). Good-quality blastocyst transfer should, therefore, be carried out irrespective of embryo quality at cleavage stage, as it may not compromise success rates in a good-prognosis population.

Predicting the clinical outcome of ICSI by sperm head vacuole examination.

To assess whether high magnification sperm head vacuole examination (SHVE) and/or standard sperm morphology assessment can predict ICSI outcomes in terms of fertilization, embryo quality, and delivery rates, a prospective observational bicentric study was conducted in two publicly funded assisted reproductive technology (ART) units in France between January and July of 2012. A total of 111 ICSI cycles for exclusively male infertility factors were included. A Spearman's correlation test was performed to validate SHVE reproducibility between the ART units. The normal morphology rate and SHVE performed on selected spermatozoa were respectively determined according to David's and Vanderzwalmen's classifications used for motile sperm organelle morphology examination (MSOME) on the day of the ICSI. Receiver Operating Characteristic (ROC) curve analysis was performed to determine thresholds associated with the occurrence of a delivery. There was an excellent correlation between the two operators (r=0.98), thus validating the study's SHVE data. Percentages of normal morphology grade spermatozoa using the standard classification and first-best morphology grade spermatozoa determined by SHVE were not significantly associated with (i) delivery (p=0.58; 0.90 /area under curve (AUC) =0.532; 0.507), (ii) fertilization (p=0.88; 0.90), (iii) top-quality embryos (p=0.27; 0.98), and (iv) good quality embryo rates (p=0.73; 0.98), respectively. In conclusion, high magnification SHVE and standard sperm morphology assessment cannot predict clinical or biological ICSI outcomes. ABBREVIATIONS: ART: assisted reproductive technology; HBV: hepatitis B virus; HCV: hepatitis C virus; HIV: human immunodeficiency virus; ICSI: intra-cytoplasmic sperm injection; IVF: in vitro fertilization; LNVs: large nuclear vacuoles; MSOME: motile sperm organelle morphology examination; SHVE: sperm head vacuole examination; WHO: World Health Organization.

Should metaphase 1 and 2 stages oocytes be vitrified in the same time for fertility preservation?

AIMS: Could metaphase 1 (M1) and 2 (M2) stages oocytes from in vitro maturation (IVM) cycles and controlled-ovarian hyperstimulation (COH) cycles be frozen at the same time without any adverse effect of vitrification on further survival (SR) and maturation rates (MR)? MATERIALS & METHODS: M1 from cancer patients were prospectively included in IVM/COH groups, and in study or control subgroups if they were vitrified or not. In each study subgroup, SR were compared with that of M2 oocytes vitrified/warmed from egg donors. MR were compared with those of fresh-M1 oocytes from control IVM/COH subgroups. RESULTS: SR were not different between groups. MR compared respectively between survived- and fresh-M1 oocytes were similar when resulting from COH (85.2 vs 81.1%) but significantly lower after IVM (39.1 vs 73.3%). CONCLUSION: Simultaneous freezing of M1/M2 oocytes could be applied to COH but not to IVM during the course of fertility preservation.

Impact on ICSI outcomes of adding 24 h of in vitro culture before testicular sperm freezing: a retrospective study.

PURPOSE: To compare sperm parameters and intracytoplasmic sperm injection (ICSI) outcomes for testicular spermatozoa frozen on the day of the biopsy (DO) with those frozen after 24 h of in vitro culture (D1). METHODS: In this retrospective study, from 1999 to 2012, forty-nine azoospermic patients were included to compare sperm (motility and viability) and outcomes (fertilization (FR), implantation (IR), pregnancy (PR) and delivery rates (DR)). RESULTS: The in vitro culture increased total motility (+2.8 %, p = 0.0161) but decreased viability (-8.3 %, p = 0.007). After 24 h of culture, the post-thaw changes in motility and viability were not significant. Twenty-six couples underwent ICSI: thirty-four ICSI were performed with spermatozoa cryopreserved at D0 and eighteen with spermatozoa frozen at D1. Cumulated IR and DR were lower for ICSI with D1 spermatozoa than with D0 spermatozoa (IR: 21.6 % with D0 vs. 9.8 % with D1, p = 0.102; DR: 27.5 % with D0 vs. 8.3 % with D1, p = 0.049). CONCLUSION: Despite improving motility, freezing spermatozoa 24 h after testicular biopsy had a potential negative effect on ICSI outcomes, notably on delivery rates. These results may be related to the detrimental impact of the additional culture on the nuclear integrity of sperm.

OBJECTIF: Comparer les paramètres spermatiques et les issues de fécondation in vitro avec micro-injection (ICSI) de spermatozoïdes testiculaires congelés le jour de la biopsie (D0) avec ceux congelés après 24 heures de culture in vitro (D1). MÉTHODES: Dans cette étude rétrospective, de 1999 à 2012, quarante-neuf patients présentant une azoospermie ont été inclus pour comparer les paramètres spermatiques (mobilité et vitalité) et les issues d'ICSI (taux de fécondation (FR), d'implantation (IR), de grossesse (PR), et d'accouchement (DR)). RÉSULTATS: La culture in vitro augmentait la mobilité (+2.8 %, p = 0.0161) mais diminuait la vitalité (-8.3 %, p = 0.007). Après cumul des 24 heures de culture et congélation, les différences observées n'étaient plus significatives. Vingt-six couples ont eu au moins une ICSI : 34 ont été réalisées avec des spermatozoïdes congelés à D0 et 18 ont été réalisées avec des spermatozoïdes congelés à D1. Les taux d'implantation et d'accouchement cumulés étaient plus faibles avec les spermatozoïdes congelés à D1 par rapport à ceux congelés à D0 (IR: 21.6 % avec D0 vs. 9.8 % avec D1, p = 0.102; DR: 27.5 % avec D0 vs. 8.3 % avec D1, p = 0.049). CONCLUSION: Malgré l'augmentation de la mobilité, la congélation de spermatozoïdes testiculaires 24 heures après la biopsie apparait avoir un impact négatif sur les issues d'ICSI, notamment sur les taux d'accouchement. Ces résultats pourraient être en lien avec les effets néfastes de l'association des deux procédés (l'incubation pendant 24H cumulée à la congélation-décongélation) sur l'intégrité nucléaire spermatique.

Could sperm grade under high magnification condition predict IMSI clinical outcome?

OBJECTIVE: The aim of this study was to examine whether injection of first-best morphology grade selected spermatozoa improves live birth rate (LBR) compared to intracytoplasmic morphologically selected sperm injection (IMSI) using second-best grade sperm. STUDY DESIGN: In this prospective observational study, 132 patients were enrolled. Inclusion criteria were the presence of severe male factor (normal spermatozoa <10% in fresh ejaculated semen and <10% in selected sperm according to David's classification) associated with ≤2 previous ICSI failure. Results of IMSI performed with either first- or second-best morphology grade spermatozoa (according to Vanderzwalmen's classification) were compared. IMSI attempts performed using mixed first- and second-best grade spermatozoa were excluded (n=41). The primary endpoint was LBR. RESULTS: LBR following IMSI was not statistically different using first- (33.3% (13/39)) or second-best morphology grade spermatozoa (28.9% (15/52)). Our study shows that sperm grading under high magnification using Vanderzwalmen's classification is not correlated to IMSI outcome. CONCLUSION: We do not validate Vanderzwalmen classification in our external and prospective series. These results point out the need for improving our knowledge about the impact of observed vacuoles under high magnification condition.

Clinical predictive criteria associated with live birth following elective single embryo transfer.

OBJECTIVE: We aimed to define clinical criteria from the patients related to the occurrence of live birth in case of elective single embryo transfer (eSET). STUDY DESIGN: We analyzed retrospectively 409 eSET at day 2/3 between March 2005 and July 2012, proposed in case of (i) woman's age <37 years, (ii) first/second IVF0 cycle, (iii) ≥2 good quality embryos obtained (3-5/6-10 blastomeres at day 2/3 and <20% fragmentation), including one top embryo (4/8 cells). In all, 124/409 live births (30.3%) were obtained, separating patients into groups of women who had birth or not. Different clinical parameters of interest were compared between each group, using appropriate statistical tests at p<0.05 significance level. RESULTS: By comparing Body Mass Index (BMI), we report a statistically higher BMI among women who did not deliver (24.6 vs. 23.4kg/m(2); p=0.014). Using an analysis by BMI categories, we also precise a threshold of BMI≥30kg/m(2), negatively associated with the occurrence of live birth. CONCLUSION: BMI appears to be the only clinical parameter statistically associated with delivery following eSET strategy in a good prognosis infertile population.