Embryo multinucleation at the two-cell stage is an independent predictor of intracytoplasmic sperm injection outcomes.

OBJECTIVE: To determine the prognostic impact of the nuclear status at the two-cell stage on intracytoplasmic sperm injection (ICSI) outcomes. DESIGN: Retrospective study. SETTING: Hospital. PATIENT(S): Only ICSI cycles with time-lapse monitoring of transferred embryos with known implantation/delivery data from November 2012 to December 2014 were included. A total of 2,449 embryos were assessed for multinucleation rates at the two- and four-cell stage, and 608 transferred embryos were studied for ICSI outcomes. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Implantation rate (IR) and live birth rate (LBR) according to the number of multinucleated blastomeres at the two-cell stage: none (Without-MNB2cell), one (MNB1/2cell), and two (MNB2/2cell); morphokinetics of MNB2cell embryos. RESULT(S): Embryos with MNB1/2cell led to lower IR (27.7%) and LBR (22.7%) than embryos Without-MNB2cell (33.4% and 29.8%, respectively). The MNB2/2cell embryos led to significantly lower IR (18.3%) and LBR (13.4%) than embryos Without-MNB2cell. This difference remained significant in multivariate analysis for implantation (odds ratio 0.57; 95% confidence interval 0.34-0.94) and birth (odds ratio 0.46; 95% confidence interval 0.26-0.80), independently of the other significant parameters (women's age, time of two-cell formation, and multinucleation at the four-cell stage). Among implanted MNB2cell, if cleavage into four cells occurred later than 37 hours after insemination, embryos were significantly more likely to lead to birth. CONCLUSION(S): The presence of multinucleation at the two-cell stage and more specifically in both blastomeres had a significant negative impact on birth potential. Thus, embryo multinucleation at the two-cell stage should be used as an additional noninvasive criterion for embryo selection.

Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?

In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium--SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05). The culture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.

6q16.3q23.3 duplication associated with Prader-Willi-like syndrome.

BACKGROUND: Prader-Willi syndrome (PWS) is characterized by hypotonia, delayed neuropsychomotor development, overeating, obesity and mental deficiency. This phenotype is encountered in other conditions, defining Prader-Willi-like syndrome (PWLS). CASE PRESENTATION: We report a 14-year-old boy with a complex small supernumerary marker chromosome (sSMC) associated with PWLS. The propositus presents clinical features commonly found in patients with PWLS, including growth hormone deficit. Banding karyotype analysis and fluorescence in situ hybridization (FISH) revealed a marker derived from chromosome 6 and a neocentromere as suspected, but array-CGH enabled us to characterize this marker as a der(10)t(6;10)(6qter → 6q23.3::10p11.1 → 10p11.21)dn. As far as we know, this is the first diagnosed case of PWLS associated with a complex sSMC, involving a 30.9 Mb gain in the 6q16.3q23.3 region and a 3.5 Mb gain in the 10p11.21p11.1 region. Several genes have been mapped to the 6q region including the TCBA1 gene, which is associated with developmental delay and recurrent infections, the ENPP1 gene, associated with insulin resistance and susceptibility to obesity and the BMIQ3 gene, associated with body mass index (BMI). No OMIM gene was found in the smallest 10p11.21p11.1 region. CONCLUSIONS: We suggest that the duplicated chromosome segment 6q16.3q23.3 may be responsible for the phenotype of our case and may also be a candidate locus of PWLS.

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.