Case reports for embryo banking: for women who want their own

Out of the many challenges in management of female factor infertility, poor responders and low response to stimulation in aged and even younger women, seems to be a common problem. It is very difficult to offer one particular management strategy or treatment protocol for optimum outcome in this group of women of poor responders. In a low resource set up, IVF (In vitro Fertilization) specialist doctors usually face a challenge in treating women with poor/ low ovarian reserve as ovum / gamete donation is considered as a taboo in various sections of society even today. Hence women insist on having an offspring of "their own" and vehemently deny ovum / gamete donations. In this article we discuss 2 cases of poor ovarian reserve retrospectively, who underwent multiple cycles of controlled ovarian hyperstimulation for embryo banking and ultimately achieved pregnancy. Both patients achieved pregnancy with the method of embryo banking. Embryo banking should be considered and discussed. Various articles have discussed the advantages and disadvantages of embryo banking or even oocytes accumulation. The advantages of this technique is patients with poor/low ovarian reserve get a chance to be pregnant with their own oocytes and also have a chance for vitrification of residual embryos. Another advantage in such patients is that the embryos can undergo PGS (Preimplantation Genetic Screening) techniques in cases of suspected genetic disorders. The disadvantage in a low resource set up like India is the cost of the treatment. Nevertheless, embryo banking and accumulation of oocytes should be given as an option for treatment of poor/ low ovarian reserve and could be considered as a ray of hope for all future mothers hoping for a child of "their own".

Effects of paternal age on human embryo development in in vitro fertilization with preimplantation genetic screening / 대한생식의학회지

OBJECTIVE: As paternal age increases, the quality of sperm decreases due to increased DNA fragmentation and aneuploidy. Higher levels of structural chromosomal aberrations in the gametes ultimately decrease both the morphologic quality of embryos and the pregnancy rate. In this study, we investigated whether paternal age affected the euploidy rate. METHODS: This study was performed using the medical records of patients who underwent in vitro fertilization (IVF) procedures with preimplantation genetic screening (PGS) from January 2016 to August 2017 at a single center. Based on their morphological grade, embryos were categorized as good- or poor-quality blastocysts. The effects of paternal age were elucidated by adjusting for maternal age. RESULTS: Among the 571 total blastocysts, 219 euploid blastocysts were analyzed by PGS (38.4%). When the study population was divided into four groups according to both maternal and paternal age, significant differences were only noted between groups that differed by maternal age (group 1 vs. 3, p=0.031; group 2 vs. 4, p=0.027). Further analysis revealed no significant differences in the euploidy rate among the groups according to the morphological grade of the embryos. CONCLUSION: Paternal age did not have a significant impact on euploidy rates when PGS was performed. An additional study with a larger sample size is needed to clarify the effects of advanced paternal age on IVF outcomes.

Freeze-all en reproducción asistida

Los avances en los protocolos de vitrificación y los resultados obtenidos tras la transferencia de embriones congelados han dado lugar a una versión distinta de los ciclos estándar de reproducción asistida: los ciclos freeze-all. Independientemente de su uso frente a las indicaciones más comunes (progesterona elevada, riesgo de hiperestimulación, entre otros), este nuevo concepto hoy representa una práctica habitual en muchas clínicas siendo aplicado a todas las pacientes. En este artículo analizaremos los distintos factores que pudieron haber contribuido a este cambio de política y la evidencia científica en relación al tema. Basados en esta evidencia concluiremos si las clínicas deberían cambiar su forma de trabajo pasando de transferencias de embriones frescos a solo transferencia de embriones congelados o si deberíamos mantener el protocolo estándar.

Breakthroughs in vitrification protocols and the results obtained after frozen embryo transfer have resulted in a different version of the assisted reproduction standard cycles: the "freeze-all" cycles. Regardless of their use beyond the usual indications (elevated progesterone, risk of hyperstimulation, among others), this new concept currently represents a common practice in many institutions and is applied to all patients. In this article, we will discuss the various factors that may have contributed to this change in policy and the scientific evidence for this topic. Based on this evidence, we will conclude if clinics should change their way of working from fresh embryo transfers to only transfer frozen embryos, or if we should maintain the standard protocol.

Analysis of clinical outcomes of different embryo stage biopsy in array comparative genomic hybridization based preimplantation genetic diagnosis and screening / 中华妇产科杂志

Objective To evaluate the efficiency of the application of array comparative genomic hybridization (array-CGH) in preimplantation genetic diagnosis or screening (PGD/PGS), and compare the clinical outcomes of different stage embryo biopsy. Methods The outcomes of 381 PGD/PGS cycles referred in the First Affiliated Hospital of Nanjing Medical University from July 2011 to August 2015 were retrospectively analyzed. There were 320 PGD cycles with 156 cleavage-stage-biopsy cycles and 164 trophectoderm-biopsy cycles, 61 PGS cycles with 23 cleavage-stage-biopsy cycles and 38 trophectoderm-biopsy cycles.Chromosomal analysis was performed by array-CGH technology combined with whole genome amplification.Single embryo transfer was performed in all transfer cycles.Live birth rate was calculated as the main clinical outcomes. Results The embryo diagnosis rate of PGD/PGS by array-CGH were 96.9%-99.1%. In PGD biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were 50.0%(58/116) and 37.2%(58/156) in cleavage-stage-biopsy group, 67.5%(85/126) and 51.8%(85/164) in trophectoderm-biopsy group (both P<0.01). In PGS biopsy cycles, the live birth rate per embryo transfer cycle and live birth rate per embryo biopsy cycle were the same as 34.8%(8/23) in cleavage-stage-biopsy group, the same as 42.1%(16/38) in trophectoderm-biopsy group (both P>0.05). Conclusions High diagnosis rate and idea live birth rate are achieved in PGD/PGS cycles based on array-CGH technology.The live birth rate of trophectoderm-biopsy group is significantly higher than that of cleavage-stage-biopsy group in PGD cycles;the efficiency of trophectoderm-biopsy is better.

Comparación de resultados posterior a la transferencia electiva de embrión único euploide en ciclos frescos vs criopreservados / Results after elective single embryo transfer: Fresh euploid embryo transfer versus frozen-thawed transfer cycles

Resumen OBJETIVO: analizar las tasas de implantación y embarazo en ciclos de fertilización in vitro con transferencia electiva de un solo blastocisto, con control del factor embriónico mediante transferencia de embriones euploides. MATERIALES Y MÉTODOS: estudio retrospectivo de pacientes atendidas entre los años 2010 a 2015 en un centro privado, en protocolo de fertilización in vitro y que tuvieron, por lo menos, un embrión euploide disponible para transferencia. Para fines de estudio las pacientes se dividieron en dos grupos: 1) transferencia de embriones frescos y 2) embriones desvitrificados. Las variables categóricas se analizaron con χ2 y prueba exacta de Fisher; las variables continuas con t de Student. Se estableció significación estadística con un valor de p < 0.05. Para el análisis estadístico se usó SAS-STAT versión 9.4. RESULTADOS: se incluyeron 637 ciclos (frescos: 243 vs criopreservados: 394). La tasa de embarazo fue de 75.5% (n = 289) vs 66.3% (n = 159), embarazo clínico 62.5% (n = 235) vs 53.1% (n = 127) que fue estadísticamente significativo a favor de los ciclos criopreservados. Las tasas de embarazo múltiple fueron bajas (1.7 vs 1.6%) en ambas cohortes. CONCLUSIONES: la transferencia de un solo embrión disminuye significativamente la incidencia de embarazos múltiples y la morbilidad materna y neonatal. El mejor pronóstico en ciclos de fertilización in vitro homólogos se consigue con la transferencia de un solo embrión genéticamente equilibrado, en un ciclo posterior de preparación endometrial sintética o natural.

Abstract OBJECTIVE: To analyze the implantation and pregnancy rates in cycles of in vitro fertilization with elective transfer of a single blastocyst, with control of the embryonic factor by transfer of euploid embryos. MATERIALS AND METHODS: Retrospective analysis who included patients that underwent IVF and had at least one euploid embryo available for transfer between 2010 and 2015 on a single academic private practice. Cohorts were segregated in fresh embryo transfers (ET) vs frozen/thawed ET. Categorical variables were analyzed with χ2 and Fisher test when appropriate. Continuous variables were analyzed with Students t test. P value < 0.5 was established as statistically significant. SAS/STAT 9.4 was used for analysis. RESULTS: Six hundred and thirty-seven euploid SETs cycles (fresh cycle: n = 243; frozen/thaw cycle: n = 394) were identified. Pregnancy (75.5% (n=289) vs 66.3% (n = 159)) and clinical pregnancy rates (PR) (62.5% (n = 235) vs 53.1% (n = 127)) were statistically higher in the frozen/thaw cycles. Low rates of multiple pregnancies (1.7 and 1.6%) were observed in both cohorts. CONCLUSIONS: In one of the largest studies to date, a euploid SET during a frozen/thaw cycle showed significantly improved pregnancy and clinical PR compared to embryo transfer in fresh cycles. Single embryo transfer significantly reduces the incidence of multiple gestation and improves maternal and neonatal outcomes. An optimal outcome is achieved by the performance of a SET in FET cycles.

Incidencia y origen parental de las aneuploidias en blastocistos: comparación entre dos centros de reproducción asistida en México / Incidence and parental origin of aneuploidies in blastocysts: comparison between two centers of assisted reproduction in Mexico

Resumen: OBJETIVO: determinar la incidencia y origen de las aneuploidias en blastocistos de dos centros mexicanos de reproducción asistida. MATERIALES Y MÉTODOS: estudio de cohorte, retrospectivo, efectuado entre los meses de enero de 2014 a diciembre de 2015 de blastocistos de día 5 y 6 obtenidos durante tratamientos de fecundación in vitro y analizados con el tamizaje genético previo a la implantación, en su variante de microarreglos de polimorfismo de nucleótido único (SNP microarrays) con el algoritmo Parental Support (Natera, USA), que permite evaluar la ploidía de los 24 cromosomas. Comparación de variables continuas: T de Student y categóricas X2. RESULTADOS: se analizaron 450 blastocistos de 80 pacientes. En el centro A: 132 blastocistos fueron de día 5 y 108 de día 6. En el centro B: 94 blastocistos fueron de día 5 y 116 de día 6. Las pacientes del centro A tuvieron mayor edad materna (37.3 ± 3.8 vs 32.4 ± 5.6; p<0.05). La incidencia total de blastocistos con aneuploidias fue similar en ambos centros; al diferenciar entre embriones de día 5 y día 6 sí hubo diferencia. El centro A reportó aumento de blastocistos aneuploides de día 6 vs blastocistos de día 5 (61.1 vs 36.3%; p<0.05). En el centro B la incidencia de embriones aneuploides fue similar entre blastocistos de día 5 y día 6 (48.9 vs 43.1; p > 0.05). El origen de las aneuploidias fue, principalmente, materno (centro A, 68.7%; centro B, 60.75%) seguido por origen mixto (centro A, 19.65%; centro B, 28.1%) y, finalmente, origen paterno (centro A, 11.6%; centro B, 11.1%). CONCLUSIONES: la incidencia de aneuploidias embrionarias entre embriones de día 5 y día 6 fue diferente entre centros. El origen fue, principalmente, materno, seguido de mixto y finalmente paterno.

Abstract: OBJECTIVE: To determine the incidence and origin of aneuploidies in blastocysts of two assisted reproduction centers in México. MATERIAL AND METHODS: Retrospective cohort study. In the period from january 2014 to December 2015, we incluided blastocysts on day 5 and day 6 of developmet, analyzed with preimplantation genetic screening; in two assisted reproduction centers. Blastocysts biopsied on day 7 and embryos that did not perform genetic diagnosis made, were excluded. The comparison of continuous variables: "T of student", categorical: X2. RESULTS: Were analized 450 blastocysts obtained from 80 patients. In center A, 132 blastocysts were on day five and 108 on day six; In the center B; 94 blastocysts were on day five and 116 on day six. Maternal age was higher in center A (37.3 ± 3.8 vs 32.4 ± 5.6 years, p <0.05). The total incidence of aneuploid blastocysts was similar in both centers; By differentiating between embryos from day five and day six if there was difference. The center A presented aneuploid blastocysts increase of day 6 compared with blastocysts of day 5 (61.1 vs 36.3%, p <0.05). In Center B the incidence of aneuploid embryos was similar between blastocysts from day 5 and 6 (48.9 vs 43.1; p> 0.05). In both centers, the main origin of aneuploidies was the maternal cause (center A, 68.7%, center B, 60.75%), followed by mixed origin (center A, 19.65%, center B, 28.1%) and finally of paternal cause (center A, 11.6%, center B, 11.1%). CONCLUSIONS: The incidence of embryonic aneuploidies between embryos from day 5 and day 6 was different between centers. The origin was mainly maternal, followed by mixed and paternal.

An update of preimplantation genetic diagnosis in gene diseases, chromosomal translocation, and aneuploidy screening / 대한생식의학회지

Preimplantation genetic diagnosis (PGD) is gradually widely used in prevention of gene diseases and chromosomal abnormalities. Much improvement has been achieved in biopsy technique and molecular diagnosis. Blastocyst biopsy can increase diagnostic accuracy and reduce allele dropout. It is cost-effective and currently plays an important role. Whole genome amplification permits subsequent individual detection of multiple gene loci and screening all 23 pairs of chromosomes. For PGD of chromosomal translocation, fluorescence in-situ hybridization (FISH) is traditionally used, but with technical difficulty. Array comparative genomic hybridization (CGH) can detect translocation and 23 pairs of chromosomes that may replace FISH. Single nucleotide polymorphisms array with haplotyping can further distinguish between normal chromosomes and balanced translocation. PGD may shorten time to conceive and reduce miscarriage for patients with chromosomal translocation. PGD has a potential value for mitochondrial diseases. Preimplantation genetic haplotyping has been applied for unknown mutation sites of single gene disease. Preimplantation genetic screening (PGS) using limited FISH probes in the cleavage-stage embryo did not increase live birth rates for patients with advanced maternal age, unexplained recurrent abortions, and repeated implantation failure. Polar body and blastocyst biopsy may circumvent the problem of mosaicism. PGS using blastocyst biopsy and array CGH is encouraging and merit further studies. Cryopreservation of biopsied blastocysts instead of fresh transfer permits sufficient time for transportation and genetic analysis. Cryopreservation of embryos may avoid ovarian hyperstimulation syndrome and possible suboptimal endometrium.

Diagnóstico genético pré-implantacional e seu valor prognóstico em tecnologia de reprodução assistida: revisão da literatura / Preimplantation genetic diagnosis and its prognostic value in assisted reproduction technology: literature review

O diagnóstico genético pré-implantacional, associado a técnicas de reprodução assistida como a fertilização in vitro, permite a seleção de pré-embriões euploides para transferência. Nos últimos anos, tem sido indicado visando o aumento nas taxas de sucesso de tratamento, na sua forma de rastreio de aneuploidias para um grupo de pacientes de alto risco reprodutivo como: idade materna avançada, aborto recorrente e falhas repetidas de tratamentos. O objetivo desta revisão é avaliar o valor prognóstico da realização do diagnóstico genético pré-implantação, associado às técnicas de fertilização in vitro e injeção intracitoplasmática de espermatozoide (ICSI), revisar as limitações da técnica e sua eficácia. Embora este grupo com elevado risco reprodutivo apresente alta taxa de aneuploidia, não foram encontrados dados consistentes na literatura a favor da realização do rastreio genético de aneuploidias; portanto, ainda são necessários estudos com grupos mais bem definidos para que o diagnóstico genético pré-implantacional possa ser confirmado como uma ferramenta para aumento de taxas ou como procedimento de rotina, até sua eficácia ser provada.(AU)

The preimplantation genetic diagnosis associated with assisted reproduction technology, as in vitro fertilization, allows selecting euploid pre-embryos for transfer. In recent years, it has been performed as a screening tool for aneuploidy with the purpose of increasing reproductive rates in patient populations at high risk of aneuploidy, such as: advanced maternal age, recurrent pregnancy loss and repeated in vitro fertilization failures. The aim of this review is to evaluate the prognostic value of preimplantation genetic diagnosis, associated with in vitro fertilization and intracytoplasmic sperm injection (ICSI) techniques, to review technology?s limitations and its effectiveness. Although this high risk population group presents high rate of aneuploidy, this review has not shown consistently data in the literature in favor of preimplantation genetic screening; therefore, more studies with well-defined groups are still needed in order that preimplantation genetic diagnosis may be confirmed as a tool for increasing rates or as a routine procedure, until technique?s problems are solved and its effectiveness, proven.(AU)