RESUMO
Among the wide range of procedures performed by clinical embryologists, the cryopreservation of reproductive cells and tissues represents a fundamental task in the daily routine. Indeed, cryopreservation procedures can be considered a subspecialty of medically assisted reproductive technology (ART), having the same relevance as sperm injection or embryo biopsy for preimplantation genetic testing. However, although a great deal of care has been devoted to optimizing cryopreservation protocols, the same energy has only recently been spent on developing and implementing strategies for the safe and reliable storage and transport of reproductive specimens. Herein, we have summarized the content of the available guidelines, the risks, the needs and the future perspectives regarding the management of cryopreservation biorepositories used in ART.
Assuntos
Técnicas de Reprodução Assistida , Sêmen , Humanos , Masculino , Células Germinativas , Criopreservação/métodos , EspermatozoidesRESUMO
PURPOSE: An impact of different gonadotrophins selection for ovarian stimulation (OS) on oocyte competence has yet to be defined. In this study, we asked whether an association exists between OS protocol and euploid blastocyst rate (EBR) per metaphase-II (MII) oocytes. METHODS: Cycles of first preimplantation genetic testing for aneuploidies conducted by women ≥ 35 years old with their own metaphase-II oocytes inseminated in the absence of severe male factor (years 2014-2018) were clustered based on whether recombinant FSH (rec-FSH) or human menopausal gonadotrophin (HMG) was used for OS, then matched for the number of fresh inseminated eggs. Four groups were outlined: rec-FSH (N = 57), rec-FSH plus rec-LH (N = 55), rec-FSH plus HMG (N = 112), and HMG-only (N = 127). Intracytoplasmic sperm injection, continuous blastocyst culture, comprehensive chromosome testing to assess full-chromosome non-mosaic aneuploidies and vitrified-warmed euploid single embryo transfers (SETs) were performed. The primary outcome was the EBR per cohort of MII oocytes. The secondary outcome was the live birth rate (LBR) per first SETs. RESULTS: Rec-FSH protocol was shorter and characterized by lower total gonadotrophin (Gn) dose. The linear regression model adjusted for maternal age showed no association between the Gn adopted for OS and EBR per cohort of MII oocytes. Similarly, no association was reported with the LBR per first SETs, even when adjusting for blastocyst quality and day of full blastulation. CONCLUSION: In view of enhanced personalization in OS, clinicians shall focus on different endpoints or quantitative effects related to Gn action towards follicle recruitment, development, and atresia. Here, LH and/or hCG was administered exclusively to women with expected sub/poor response; therefore, we cannot exclude that specific Gn formulations may impact patient prognosis in other populations.
Assuntos
Gonadotropinas , Sêmen , Masculino , Feminino , Humanos , Adulto , Estudos de Casos e Controles , Idade Materna , Metáfase , Gonadotropinas/uso terapêutico , Gonadotropinas/farmacologia , Oócitos , Indução da Ovulação/métodos , Menotropinas/uso terapêutico , Hormônio Foliculoestimulante/uso terapêutico , Hormônio Foliculoestimulante/farmacologia , Aneuploidia , Fertilização in vitroRESUMO
PURPOSE: To investigate whether high relative humidity conditions (HC), when using a time-lapse system (TLS) with sequential culture media, are beneficial to embryo culture, improving ongoing pregnancy rates. METHODS: We included patients undergoing their first ICSI cycle treatment from April 2021 to May 2022. Patients assigned to dry conditions (DC) or HC were 278 and 218, respectively. We used a GERI TLS, three chambers configured in humidity conditions and three in dry conditions. The effect of HC on ongoing pregnancy rate was assessed by the propensity matched sample, to reduce potential differences between women undergoing either HC or DC and reduce biased estimation of treatment effect. RESULT: After adjusting for several confounding variables and applying the propensity score (PS), no significant differences were observed in the rates of normal (2PN) and abnormal (1PN and 3PN) fertilization, blastulation, top-quality blastocysts, frozen blastocysts, ongoing pregnancies, and miscarriages. The 2-cell (t2) and 4-cell (t4) stages and cell divisions between such stages occurred earlier and were more synchronous in the in DC. CONCLUSION: These results suggest that HC conditions do not improve the rate of ongoing pregnancy and several embryological outcomes, under the conditions used in this study based on a time-lapse system and sequential culture with day 3 medium change-over.
Assuntos
Desenvolvimento Embrionário , Fertilização in vitro , Gravidez , Humanos , Feminino , Taxa de Gravidez , Estudos Retrospectivos , Fertilização in vitro/métodos , Injeções de Esperma Intracitoplásmicas/métodos , Imagem com Lapso de Tempo , Pontuação de Propensão , Blastocisto , Técnicas de Cultura Embrionária/métodosRESUMO
RESEARCH QUESTION: What is the intra- and inter-centre reliability in embryo grading performed according to the Istanbul Consensus across several IVF clinics? DESIGN: Forty Day 3 embryos and 40 blastocysts were photographed on three focal planes. Senior and junior embryologists from 65 clinics were invited to grade them according to the Istanbul Consensus (Study Phase I). All participants then attended interactive training where a panel of experts graded the same embryos (Study Phase II). Finally, a second set of pictures was sent to both embryologists and experts for a blinded evaluation (Study Phase III). Intra-centre reliability was reported for Study Phase I as Cohen's kappa between senior and junior embryologists; inter-centre reliability was instead calculated between senior/junior embryologists and experts in Study Phase I versus III to outline improvements after training (i.e. upgrade of Cohen's kappa category according to Landis and Koch). RESULTS: Thirty-six embryologists from 18 centres participated (28% participation rate). The intra-centre reliability was (i) substantial (0.63) for blastomere symmetry (range -0.02 to 1.0), (ii) substantial (0.72) for fragmentation (range 0.29-1.0), (iii) substantial (0.66) for blastocyst expansion (range 0.19-1.0), (iv) moderate (0.59) for inner cell mass quality (range 0.07-0.92), (v) moderate (0.56) for trophectoderm quality (range 0.01-0.97). The inter-centre reliability showed an overall improvement from Study Phase I to III, from fair (0.21-0.4) to moderate (0.41-0.6) for all parameters under analysis, except for blastomere fragmentation among senior embryologists, which was already moderate before training. CONCLUSIONS: Intra-centre reliability was generally moderate/substantial, while inter-centre reliability was just fair. The interactive training improved it to moderate, hence this workflow was deemed helpful. The establishment of external quality assessment services (e.g. UK NEQAS) and the avant-garde of artificial intelligence might further improve the reliability of this key practice for embryo selection.
Assuntos
Inteligência Artificial , Blastocisto , Embrião de Mamíferos , Fertilização in vitro , Humanos , Reprodutibilidade dos TestesRESUMO
RESEARCH QUESTION: The study aimed to retrospectively evaluate the impact of cryo-storage duration on clinical, obstetric and perinatal outcomes after vitrified-warmed euploid blastocyst transfer. DESIGN: This was an observational study including 2688 vitrified-warmed euploid single blastocyst transfers that was conducted at a private IVF centre between May 2013 and March 2020. It included a total of 1884 women (age 38 ± 3 years) undergoing at least one transfer after preimplantation genetic testing for aneuploidies. The euploid blastocysts transferred were clustered into seven groups according to the cryo-storage duration between vitrification and warming: ≤60 days (nâ¯=â¯646; control group), 61-90 days (nâ¯=â¯599), 91-180 days (nâ¯=â¯679), 181-360 days (nâ¯=â¯405), 361-720 days (nâ¯=â¯144), 721-1080 days (nâ¯=â¯118) and >1080 days (nâ¯=â¯97). The primary outcome was the live birth rate (LBR) per transfer. The secondary outcomes were miscarriage rate, obstetric and perinatal issues. The data were adjusted for confounders through logistic or linear regressions. RESULTS: A significantly lower LBR was reported for transfers performed within 91-180 days (nâ¯=â¯291/679, 42.9%; Pâ¯=â¯0.017), 181-360 days (nâ¯=â¯169/405, 41.7%; Pâ¯=â¯0.016) and 361-720 days (nâ¯=â¯57/144, 39.6%; Pâ¯=â¯0.034) versus ≤60 days (nâ¯=â¯319/646, 49.4%). However, this was mainly due to top-quality embryos being transferred first when more euploid blastocysts were available, thereby leaving lower quality ones for subsequent procedures. Indeed, the multivariate odds ratios adjusted for confounders showed similar results across all cryo-storage duration clusters. No difference was reported also for all secondary outcomes. CONCLUSIONS: Cryo-storage duration even beyond 3 years from blastocyst vitrification does not affect clinical, obstetric and perinatal outcomes.
Assuntos
Transferência Embrionária , Vitrificação , Adulto , Aneuploidia , Blastocisto , Criopreservação/métodos , Transferência Embrionária/métodos , Feminino , Humanos , Masculino , Gravidez , Taxa de Gravidez , Estudos RetrospectivosRESUMO
The goal of an IVF cycle is the birth of at least one baby per intention to treat. However, IVF cannot confer competence on an embryo, but only can provide each couple with a safe treatment to meet a predetermined chance of success. This commentary highlights how clinical, financial and patient-centred perspectives should be included in the definition of success in IVF. The primary outcome, which is the cumulative live birth delivery rate per intention to treat, must always be complemented by analyses of risks, costs and time invested, as well as by measures of patient satisfaction. Finally, it is essential, whenever clinical conditions exist, to limit treatment discontinuation after failed attempts. Constant monitoring of the data is pivotal and must be adjusted for patient characteristics and compared with national and international registers. The authors aimed to review all these aspects and highlight the points that are still open for discussion. Is it time for a consensus?
Assuntos
Consenso , Fertilização in vitro , Comunicação Interdisciplinar , Resultado do Tratamento , Análise Custo-Benefício , Aconselhamento , Feminino , Fertilização in vitro/economia , Fertilização in vitro/psicologia , Humanos , Nascido Vivo , Masculino , Satisfação do Paciente , Gravidez , Fatores de RiscoRESUMO
RESEARCH QUESTION: An evidence-based novel commercially available continuous IVF culture medium in compliance with an efficient quality-management system is proposed. DESIGN: Non-interventional study on sibling oocytes. Intracytoplasmic sperm injection cycles among women aged 42 years or younger that used ejaculated spermatozoa and retrieved four to eight oocytes were included. Sibling oocytes were randomized for culture in the novel Geri-medium or continuous single culture medium (CSCM). Primary outcome measure was blastulation rate per cohort of inseminated oocytes; 1182 oocytes were required to outline down to a 7% difference (powerâ¯=â¯80%). RESULTS: A total of 181 cohorts of sibling oocytes were included. Geri-medium (nâ¯=â¯631 oocytes) and CSCM (nâ¯=â¯643 oocytes) resulted in similar blastulation rates (mean ± SD: 42.8% ± 30.1% versus 43.1% ± 29.0%; Wilcoxon signed rank testâ¯=â¯0.77). Blastocysts cultured in the former (nâ¯=â¯275 versus nâ¯=â¯277) showed longer timings during preimplantation development (P < 0.01) and were poorer quality (26% versus 18%; Pâ¯=â¯0.03). Euploidy rate was no different in cycles that underwent preimplantation genetic testing for aneuploidy (nâ¯=â¯113) (117/237 [49%] versus 117/249 blastocysts [47%]; Pâ¯=â¯0.6). Ongoing implantation rate was comparable in the study arms after euploid (29/47 [63%] versus 14/ 34 [41%]; Pâ¯=â¯0.1) or untested (12/31 [39%] versus 7/18 [39%]; Pâ¯=â¯0.3) transfers. CONCLUSION: Blastulation rate among cohorts of sibling oocytes cultured in the same incubator is a fast, reliable and comprehensive performance indicator to validate novel commercially available culture medium. The media tested were considered similarly efficient. The differences in blastocyst morphology and developmental timings warrant further investigation, although euploidy and ongoing implantation rates were similar.
Assuntos
Meios de Cultura/farmacologia , Técnicas de Cultura Embrionária/estatística & dados numéricos , Desenvolvimento Embrionário/efeitos dos fármacos , Adulto , Feminino , Humanos , Masculino , Fluxo de TrabalhoRESUMO
RESEARCH QUESTION: Does maternal preconceptional body mass index (BMI) associate with mean blastocyst euploidy rate (m-ER) per patient and live birth rate (LBR) after vitrified-warmed euploid single embryo transfer (SET)? DESIGN: Observational study conducted between April 2013 and March 2020 at a private IVF clinic, involving 1811 Caucasian women undergoing trophectoderm biopsy and comprehensive chromosome testing. The outcomes of 1125 first vitrified-warmed euploid SET were also analysed. Patients were clustered as normal weight (BMI 18.5-25; nâ¯=â¯1392 performing 859 SET), underweight (BMI <18.5; nâ¯=â¯160 performing 112 SET) and overweight (BMI >25; nâ¯=â¯259 performing 154 SET). m-ER per patient was the primary outcome. The secondary outcomes were all clinical outcomes per euploid SET. All data were adjusted for confounders through regression analyses. RESULTS: The m-ER per patient decreases as maternal BMI increases from 17 up to 22-23 before reaching a plateau. A linear regression adjusted for maternal age confirmed this moderate association (unstandardized coefficient B: -0.6%, 95% confidence interval [CI]: -1.1 to -0.1%, Pâ¯=â¯0.02). All clinical outcomes were similar between normal weight and underweight women. Overweight women, instead, showed higher miscarriage rate per clinical pregnancy (nâ¯=â¯20/75, 26.7% versus nâ¯=â¯67/461, 14.5%; odds ratio [OR] adjusted for blastocyst quality and day of full blastulation: 2.0, 95% CI: 1.1-3.6, Pâ¯=â¯0.01) and lower LBR per SET (nâ¯=â¯55/154, 35.7% versus nâ¯=â¯388/859, 45.2%; OR adjusted for blastocyst quality and day of full blastulation: 0.67, 95% CI: 0.46-0.96, Pâ¯=â¯0.03). CONCLUSION: These data indicate a need for future research on more sensitive metrics to assess body fat mass and distribution, as well as on the mechanisms leading to lipotoxicity, thereby impairing embryo competence and/or endometrial receptivity. Overweight women should be informed of their higher risk for miscarriage and, whenever possible, encouraged to lose weight, especially before transfer.
Assuntos
Aborto Espontâneo/etiologia , Coeficiente de Natalidade , Índice de Massa Corporal , Embrião de Mamíferos/anormalidades , Sobrepeso/complicações , Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Gravidez , Estudos Retrospectivos , Adulto JovemRESUMO
PURPOSE: To assess whether the GnRH-agonist or urinary-hCG ovulation triggers affect oocyte competence in a setting entailing vitrified-warmed euploid blastocyst transfer. METHODS: Observational study (April 2013-July 2018) including 2104 patients (1015 and 1089 in the GnRH-a and u-hCG group, respectively) collecting ≥1 cumulus-oocyte-complex (COC) and undergoing ICSI with ejaculated sperm, blastocyst culture, trophectoderm biopsy, comprehensive-chromosome-testing, and vitrified-warmed transfers at a private clinic. The primary outcome measure was the euploid-blastocyst-rate per inseminated oocytes. The secondary outcome measure was the maturation-rate per COCs. Also, the live-birth-rate (LBR) per transfer and the cumulative-live-birth-delivery-rate (CLBdR) among completed cycles were investigated. All data were adjusted for confounders. RESULTS: The generalized-linear-model adjusted for maternal age highlighted no difference in the mean euploid-blastocyst-rate per inseminated oocytes in either group. The LBR per transfer was similar: 44% (n=403/915) and 46% (n=280/608) in GnRH-a and hCG, respectively. On the other hand, a difference was reported regarding the CLBdR per oocyte retrieval among completed cycles, with 42% (n=374/898) and 25% (n=258/1034) in the GnRh-a and u-hCG groups, respectively. Nevertheless, this variance was due to a lower maternal age and higher number of inseminated oocytes in the GnRH-a group, and not imputable to the ovulation trigger itself (multivariate-OR=1.3, 95%CI: 0.9-1.6, adjusted p-value=0.1). CONCLUSION: GnRH-a trigger is a valid alternative to u-hCG in freeze-all cycles, not only for patients at high risk for OHSS. Such strategy might increase the safety and flexibility of controlled-ovarian-stimulation with no impact on oocyte competence and IVF efficacy.
Assuntos
Gonadotropina Coriônica/genética , Fertilização in vitro , Hormônio Liberador de Gonadotropina/genética , Oócitos/crescimento & desenvolvimento , Adulto , Coeficiente de Natalidade , Blastocisto/metabolismo , Gonadotropina Coriônica/metabolismo , Técnicas de Cultura Embrionária/tendências , Transferência Embrionária/tendências , Feminino , Hormônio Liberador de Gonadotropina/agonistas , Humanos , Nascido Vivo/epidemiologia , Recuperação de Oócitos , Oócitos/transplante , Ovulação/genética , Indução da Ovulação/métodos , Gravidez , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas , VitrificaçãoRESUMO
STUDY QUESTION: Do the ICSI-related procedural timings and operators affect the outcomes of an ART cycle? SUMMARY ANSWER: The ICSI-related timings and operators do not associate with the mean blastulation rate per cohort of inseminated oocytes and the cumulative delivery rate per concluded cycle, except for a mild association between the times from induction of ovulation to oocyte denudation and the former outcome. WHAT IS KNOWN ALREADY: In ART, specific timings, protocols and conditions must be complied with to preserve gamete developmental and reproductive competence during the required manipulations. ICSI represents a groundbreaking advancement that has been widely implemented. Nevertheless, the studies that examined the putative impact of ICSI-related procedural timings were mainly conducted in old-fashioned settings or in good prognosis patients. No report addressed issues like operators' skills and experience and uncertainties exist dealing with the effect of cumulus cells in the pre-incubation period in vitro before ICSI. However, all this information is crucial to efficiently plan the daily routine of an IVF lab, fill the existing gaps of knowledge and define proper key performance indicators. STUDY DESIGN, SIZE, DURATION: Observational study conducted at a private IVF clinic (January 2016 to January 2018). We included all consecutive ICSI procedures (n = 1084 infertile couples undergoing 1444 cycles with or without preimplantation genetic testing (PGT); mean ± SD maternal age: 38.1 ± 4.0 years) with fresh autologous oocytes (n = 7999 oocytes, 5.5 ± 3.2 per treatment) inseminated with fresh non-donor ejaculated sperm. All operators and critical procedural timings (induction of ovulation to oocyte denudation, denudation and ICSI) were automatically recorded through an electronic witnessing system. The primary outcome measure was the cumulative delivery rate among both non-PGT and PGT-concluded cycles (i.e. delivery achieved or no supernumerary cryopreserved blastocyst available). The secondary outcome measure was the mean blastulation rate per cohort of inseminated oocytes. All confounders were registered and included in generalized linear models and multivariate logistic regression analyses. PARTICIPANTS/MATERIALS, SETTING, METHODS: Fourteen and 12 operators were involved in denudation and ICSI procedures, respectively. Denudation was performed after 4.1 ± 1.2 h (2-7) of pre-incubation in vitro after oocyte retrieval, and ICSI was started immediately after. Beyond procedural timings and operators, all the putative confounders (patients' and cycles' characteristics) on the primary and/or secondary outcomes were systematically registered and included in the statistical analyses. MAIN RESULTS AND THE ROLE OF CHANCE: The mean time from induction of ovulation to oocyte denudation was 39.3 ± 1.3 h. The mean procedural timings for denudation and ICSI were 8.1 ± 3.8 and 12.6 ± 6.4 min; both these variables were significantly dependent on the number of inseminated oocytes and the operators' skills and experience. The overall mean blastulation rate per cohort of inseminated oocytes was 34.0 ± 27.9%. This outcome was significantly associated with the time from induction of ovulation to oocyte denudation (mean blastulation rate stable in the time interval 38-42 h, but significantly higher for timings <38 h), maternal age (the mean blastulation rate drops especially beyond the age of 40 years) and categorized sperm concentration (highest mean blastulation rate for sperm concentrations ≥15 mil/ml and lowest for cryptozoospermic patients) through a generalized linear model that showed an adjusted r2 = 0.053 (P < 0.01). No association was found for denudation and ICSI timings and operators. Lastly, when adjusted for maternal age and number of inseminated oocytes, both ICSI-related procedural timings and operators did not associate with the cumulative delivery rate among both non-PGT- or PGT-concluded cycles. LIMITATIONS, REASONS FOR CAUTION: This is a single private IVF center study. Its reproducibility should be assessed in different laboratory conditions, with different protocols and in the hands of different operators. Moreover, specific studies are warranted to address the beneficial/detrimental effect of the other putative confounders under investigation (e.g. kind of ovulation trigger, culture media, incubator, etc.). WIDER IMPLICATIONS OF THE FINDINGS: Proactive communication between the embryologists and the clinicians might contribute to a reasoned and more efficient organization of the daily workload and increase the mean blastulation rate, especially when poor prognosis couples (advanced maternal age, reduced sperm count and/or ovarian reserve) are treated. STUDY FUNDING/COMPETING INTEREST(S): No funding. The authors declare no conflict of interest related to the present study.
Assuntos
Infertilidade , Injeções de Esperma Intracitoplásmicas , Adulto , Feminino , Fertilização in vitro , Humanos , Masculino , Recuperação de Oócitos , Indução da Ovulação , Gravidez , Taxa de Gravidez , Reprodutibilidade dos Testes , Estudos RetrospectivosRESUMO
RESEARCH QUESTION: The study set out to identify corrective measures aimed at reducing the risk of aerosol-mediated viral infection within an IVF laboratory. DESIGN: A failure modes and effect analysis (FMEA) was conducted by a multidisciplinary IVF team. A schematic representation of new protocols and procedures adopted during COVID-19 emergency has been defined, including directives about the behaviour to adopt when entering the clinic and the laboratory, in case of face-to-face contact with patients and between staff members. In addition, the risk of cross-contamination between samples belonging to different patients during cell handling and manipulation has been evaluated. Potential failure modes for each phase of the emergency have been analysed, focusing on possible sources of error. Risk priority numbers have been calculated as products of Occurrenceâ¯×â¯Severityâ¯×â¯Detection scores. RESULTS: Except for cell-cell contamination, which was considered highly unlikely, failure modes during patient-staff, staff-staff and staff-cell interactions were estimated as carrrying a moderate to high risk of infection. The main corrective measures entailed precautionary logistic measures, the implementation of additional personal protective equipment and changes in the IVF laboratory procedures and scheduling of the daily routine. Some procedures were also revised, aiming to increase staff's awareness and caution. CONCLUSIONS: Standard laboratory protocols are insufficient to face a virus whose transmission is aerosol mediated. The measures outlined in this FMEA should thus be considered not only for facing this pandemic, but also for the future to promptly manage any aerosol-mediated virus infection, whose impact on the management of an IVF laboratory might be less severe than COVID-19 although not completely negligible.
Assuntos
Betacoronavirus , Infecções por Coronavirus/prevenção & controle , Fertilização in vitro/métodos , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/transmissão , Humanos , Programas de Rastreamento , Pessoal de Laboratório Médico , Pacientes , Equipamento de Proteção Individual , Pneumonia Viral/diagnóstico , Pneumonia Viral/transmissão , Medição de Risco , Fatores de Risco , SARS-CoV-2 , TelemedicinaRESUMO
The term "cryopreservation" refers to the process of cooling cells and tissues and storing them at subzero temperatures in order to stop all biological activity and preserve their viability and physiological competences for future use. Cooling to subzero temperatures is not a physiological condition for human cells; this is probably due to the high content of water in the living matter, whose conversion to ice crystals may be associated with severe and irreversible damage. Among reproductive cells and tissues, metaphase II oocytes are notably vulnerable to cryopreservation, mainly because of their large size, low surface area to volume ratio, relatively high water content and presence of the meiotic spindle. As human biological systems lack efficient internal defense mechanisms against chilling injuries, it is of the utmost importance to supply adequate external support, in terms of cryoprotectant additives, appropriate cooling/warming rates, and suitable long-term storage. Over the years, scientists have proposed different cryopreservation strategies in the effort to achieve an optimized recipe ensuring cell survival and, at the same time, maintenance of the physiological functions and abilities necessary to continue life. However, despite the first success obtained in the 1980s with frozen oocytes, it was not until recently that notable improvements in the cryopreservation technique, thanks to the advent of vitrification, allowed a breakthrough of this fine procedure.
Assuntos
Criopreservação , Oócitos , Sobrevivência Celular , Criopreservação/métodos , Criopreservação/tendências , Feminino , Preservação da Fertilidade/métodos , Humanos , VitrificaçãoRESUMO
SummaryThe aim of the present study was to determine whether clinical or laboratory factors can influence the development of single pronucleated zygotes (1PN) and two polar bodies (PB) after ICSI. In total, 341 ICSI cycles performed at FertiClinic-Villa Margherita from January 2012 to December 2014 were enrolled in the study. Group A included 240 cycles with no 1PN-2PB while group B included 101 cycles with one or more 1PN-2PB. Age, stimulation protocol, infertility factor, amount of gonadotropin administered, duration of therapy, peak estradiol levels, number of follicles at maturation triggering, oocytes retrieved and mature oocytes, time between retrieval and injection and sperm characteristics were compared between groups. In opposition to previous results showing no relationship between 1PN occurrence and clinical or laboratory variables, we observed that 1PN-2PB zygote formation seems to be associated with a lower female age, higher level of E2 and higher number of follicles on day of oocyte maturation triggering, higher number of astenozoospermic male patients, more oocytes retrieved at pick-up, more mature oocytes (MII) and longer time to injection.
Assuntos
Infertilidade Masculina/patologia , Injeções de Esperma Intracitoplásmicas/métodos , Zigoto/citologia , Zigoto/fisiologia , Adulto , Estradiol/metabolismo , Feminino , Humanos , Masculino , Análise Multivariada , Recuperação de Oócitos , Corpos Polares , Estudos Retrospectivos , Espermatozoides/fisiologiaRESUMO
The aim of this study was to describe a system for embryo morphology scoring at the morula stage and to determine the efficiency of this model in selecting viable embryos for transfer. In total, 519 embryos from 122 patients undergoing intracytoplasmic sperm injection (ICSI) were scored retrospectively on day 4 according to the grading system proposed in this article. Two separate quality scores were assigned to each embryo in relation to the grade of compaction and fragmentation and their developmental fate was then observed on days 5 and 6. Secondly, the prediction value of this scoring system was compared with the prediction value of the traditional scoring system adopted on day 3. Morulas classified as grade A showed a significant higher blastocyst formation rate (87.2%) compared with grades B, C and D (63.8, 41.3 and 15.0%, respectively), (P < 0.001). Furthermore, the ability to form top quality blastocysts was significantly higher for grade A morulas with respect to grades B, and C and D (37.8% vs. 22.4% vs. 11.1%), (P < 0.001). Finally, the morula scoring system showed more prediction power with respect to the embryo scoring a value of 1 [Akaike information criterion (AIC) index 16.4 vs. 635.3 and Bayesian information criterion (BIC) index -68.8 vs. -30.0 for morulas and embryos respectively]. In conclusion, results demonstrated that the presented scoring system allows for the evaluation of eligible embryos for transfer as a significant correlation between the grade of morula, blastulation rate and blastocyst quality was observed. Furthermore, the morula scoring system was shown to be the best predictive model when compared with the traditional scoring system performed on day 3.
Assuntos
Blastocisto/fisiologia , Transferência Embrionária/métodos , Desenvolvimento Embrionário/fisiologia , Fertilização in vitro/métodos , Mórula/fisiologia , Injeções de Esperma Intracitoplásmicas/métodos , Adulto , Implantação do Embrião , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Fatores de TempoRESUMO
PURPOSE: The aim of the present randomized, comparative study was to evaluate the effect of reduced culture volumes on sibling human embryo development. METHODS: Firstly, sibling injected oocytes obtained from 88 out of 165 consenting couples undergoing infertility treatment were cultured either in large (35 µl) or in small drops (15 µl) of culture medium. Secondly, sibling injected oocytes from 77 couples were cultured either in large (35 µl) or in mini drops (7 µl). Embryo quality on day-2 and day-3 and blastocyst formation rate on day-5 were evaluated. RESULTS: No statistically significant difference in terms of embryo quality was detected comparing embryos cultured either in large (35 µl) or small (15 µl) drops until blastocyst stage. Similarly, no difference appeared between large (35 µl) or mini (7 µl) drops until day-3, however a significantly higher blastocyst formation rate was observed in mini (7 µl) drops on day-5. CONCLUSIONS: Reduced culture volume seems not to influence early embryo development but a reduction of medium appears to positively affect blastocyst development. This supports the hypothesis that the pre-implantation embryo produces autocrine factors which exert a positive effect on embryo development when culture is performed in a reduced volume.
Assuntos
Blastocisto/fisiologia , Técnicas de Cultura Embrionária/métodos , Adulto , Blastocisto/citologia , Meios de Cultura , Feminino , Humanos , Masculino , Oócitos/fisiologiaRESUMO
The capability of human zona pellucida (ZP) to bind selectively to normal functional sperm with normal chromatin has been reported widely in the literature. The aim of this study was to evaluate whether ZP-binding sperm selection may represent a method to retrieve superior spermatozoa for intracytoplasmic sperm injection (ICSI). Patients were divided into two groups: a ZP-ICSI and a conventional ICSI group. In the ZP-ICSI group, spermatozoa for injection were selected after ZP-sperm incubation and spermatozoa that were tightly bound to the ZP were used for ICSI (ZP-ICSI). Clinical outcomes of ZP-ICSI were compared with the outcomes of traditional scientist-selected sperm injection (conventional ICSI). Results did not show any significant difference in fertilization, pregnancy, implantation and take-home-baby rates between conventional ICSI and ZP-ICSI. However, when data relative to patients who received ZP-ICSI were analyzed, an interesting result was observed: higher sperm concentration and morphology correlated with higher ZP-sperm binding. Additionally, patients with higher ZP-sperm binding seem to have improved pregnancy and take-home-baby rates. In conclusion, this study shows that ZP-ICSI is not a superior method compared with conventional ICSI. However, clinical ICSI outcomes were apparently improved in the presence of good ZP-sperm binding. We therefore speculate that sperm competence to ICSI could be reduced when the sperm's ability to bind the ZP is impaired.
Assuntos
Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo , Adulto , Feminino , Humanos , Infertilidade Masculina/patologia , Masculino , Gravidez , Taxa de Gravidez , Interações Espermatozoide-ÓvuloRESUMO
OBJECTIVE: To assess whether high body mass index (BMI) in either oocyte donors or recipients is associated with poorer outcomes after the first single blastocyst transfer. DESIGN: Retrospective study including 1,394 first blastocyst single embryo transfers (SETs) conducted by 1,394 recipients during oocyte donation cycles with the gametes retrieved from 1,394 women (January 2019-July 2021). Four BMI clusters were defined for both donors and recipients (underweight: <18.5 kg; normal weight: 18.5-24.9 kg; overweight: 25-29.9 kg; and obese: ≥30 kg). SETTING: Network of private IVF centers. PATIENTS: A total of 1,394 recipients aged 42.4 ± 4.0 and with a BMI of 23.2 ± 3.8 kg/m2, and 1,394 donors aged 26.1 ± 4.2 and with a BMI of 21.9 ± 2.5 kg/m2. INTERVENTION: All oocytes were vitrified at 2 egg banks and warmed at 8 in vitro fertilization clinics that were part of the same network. Intracytoplasmic sperm injection, blastocyst culture, and either fresh or vitrified-warmed SETs were conducted. Putative confounders were investigated, and the data were adjusted through regression analyses. MAIN OUTCOME MEASURES: The primary outcome was the live birth rate (LBR) per SET according to donors' and/or recipients' BMI. The main secondary outcome was the miscarriage rate (<22 gestational weeks) per clinical pregnancy. RESULTS: The LBR per blastocyst SET showed no significant association with donors' BMI. Regarding recipients' BMI, instead, the multivariate odds ratio was significant in obese vs. normal-weight recipients (0.58, 95% confidence interval, 0.37-0.91). The miscarriage rate per clinical pregnancy was also significantly associated with recipients' obesity, with a multivariate odds ratio of 2.31 (95% confidence interval, 1.18-4.51) vs. normal-weight patients. A generalized additive model method was used to represent the relationship between predicted LBR or miscarriage rates and donors' or recipients' BMI; it pictured a scenario where the former outcome moderately but continuously decreases with increasing recipients' BMI to then sharply decline in the BMI range of 25-35 kg/m2. The miscarriage rate, instead, increases almost linearly with respect to both donors' and recipients' increasing BMI. CONCLUSION: Obesity mostly affects the uterus, especially because of higher miscarriage rates. Yet, poorer outcomes can be appreciated already with a BMI of 25 kg/m2 in both oocyte donors and recipients. Finer markers of nutritional homeostasis are therefore desirable; recipients should be counseled about poorer expected outcomes in cases of overweight and obesity; and oocyte banks should avoid assigning oocytes from overweight donors to overweight and obese recipients.
Assuntos
Aborto Espontâneo , Gravidez , Humanos , Masculino , Feminino , Índice de Massa Corporal , Aborto Espontâneo/epidemiologia , Aborto Espontâneo/etiologia , Estudos Retrospectivos , Taxa de Gravidez , Sobrepeso/diagnóstico , Sobrepeso/epidemiologia , Sobrepeso/terapia , Sêmen , Transferência Embrionária/efeitos adversos , Fertilização in vitro/efeitos adversos , Útero , Obesidade/diagnóstico , Obesidade/epidemiologia , Obesidade/terapia , Oócitos , BlastocistoRESUMO
The aim of our study was to retrospectively evaluate whether the oral administration of L. crispatus (M247) could increase pregnancy and live birth rates in women undergoing assisted reproductive technology procedures. Enrolled women (N = 160) were divided into two groups: treated (N = 80) or untreated (N = 80) with the probiotic strain. The odds ratio (OR) for a treated woman to have a clinical pregnancy (CP) was 1.56. In women aged 30-40 years, M247 increased the probability of a CP in correlation with the progressive rise in BMI, reaching 47% (35% in controls) with a BMI of 35 (OR: 2.00). The CAID statistics showed that in a woman of the blastocyst subgroup, below 43 years, with a BMI over 18.6, treatment with M247 increased the chance of a CP from 28.4% to 44.5% (OR: 2.08; p < 0.05). Considering live births, the rate of the probiotic group was 12.5% versus 7.5% (OR: 1.76). Considering only the blastocyst subgroup, the treatment increased the number of live births by 200% (OR: 3.64; p = 0.05). As confirmed also by statistical indices NNT, NNH, and LHH, the use of M247 demonstrated a risk-benefit ratio to the full advantage of the benefits.
RESUMO
Environmental toxicants (ETs) are an exogenous chemical group diffused in the environment that contaminate food, water, air and soil, and through the food chain, they bioaccumulate into the organisms. In mammals, the exposure to ETs can affect both male and female fertility and their reproductive health through complex alterations that impact both gametogeneses, among other processes. In humans, direct exposure to ETs concurs to the declining of fertility, and its transmission across generations has been recently proposed. However, multi- and transgenerational inheritances of ET reprotoxicity have only been demonstrated in animals. Here, we review recent studies performed on laboratory model animals investigating the effects of ETs, such as BPA, phthalates, pesticides and persistent contaminants, on the reproductive system transmitted through generations. This includes multigenerational effects, where exposure to the compounds cannot be excluded, and transgenerational effects in unexposed animals. Additionally, we report on epigenetic mechanisms, such as DNA methylation, histone tails and noncoding RNAs, which may play a mechanistic role in a nongenetic transmission of environmental information exposure through the germline across generations.
Assuntos
Histonas , Praguicidas , Animais , Feminino , Humanos , Masculino , Mamíferos/genética , Reprodução , Solo , Poluição Química da ÁguaRESUMO
The gut microbiota (GM) is a complex and dynamic population of microorganisms living in the human gastrointestinal tract that play an important role in human health and diseases. Recent evidence suggests a strong direct or indirect correlation between GM and both male and female fertility: on the one hand, GM is involved in the regulation of sex hormone levels and in the preservation of the blood-testis barrier integrity; on the other hand, a dysbiotic GM is linked to the onset of pro-inflammatory conditions such as endometriosis or PCOS, which are often associated with infertility. Exposure to endocrine-disrupting chemicals (EDCs) is one of the main causes of GM dysbiosis, with important consequences to the host health and potential transgenerational effects. This perspective article aims to show that the negative effects of EDCs on reproduction are in part due to a dysbiotic GM. We will highlight (i) the link between GM and male and female fertility; (ii) the mechanisms of interaction between EDCs and GM; and (iii) the importance of the maternal-fetal GM axis for offspring growth and development.