Time-Lapse Embryo culture: A better understanding of embryo development and clinical application.

Conventional embryo assessment is performed by removing embryos from incubators at least once a day. However, it is static and limited to specific time points, reducing the amount of information that could potentially be obtained. Fortunately, the time-lapse system is a powerful technology that enables to observe embryo development progression by image acquisition at recurrent time intervals, without interfering in the culture conditions. There are numerous studies that used time-lapse incubators, focusing on embryo kinetics, patient characteristics and clinical outcomes. This review aims to find agreements in the literature concerning embryo kinetics and patient characteristics: age, body mass index, smoking habit, polycystic ovary syndrome and endometriosis; as well as culture conditions that involved culture media and oxygen concentration. Furthermore, they showed differences according to ploidy status, direct/reverse cleavage, gender and the potential association between embryo collapse and clinical outcomes.

What advice should we give our patients to preserve their fertility and avoid needing oocyte donation in the future? - A Social Fertility Preservation program.

OBJECTIVE: To describe our fertility preservation program focusing on the number of oocytes vitrified by age. METHODS: From January 2015 to December 2016, 686 oocyte vitrification cycles were performed in our units for the social fertility preservation program. In total, 288 were donors who donated their oocytes for our oocyte-banking program, and 398 were patients who underwent elective fertility preservation. RESULTS: The mean numbers of COCs retrieved and vitrified oocytes were similar among the donor cycles (women under 30 years). In those patients over 36 years of age the mean numbers of COCs retrieved and vitrified oocytes were significantly lower. We also estimated the association between age and cancelation rates. Odd ratios (OR) for total cancelation was calculated between patients of 31-35 years and 41-45 years; the OR was 5.17 (95% CI 1.89 - 14.17) and increased up to 25.67 (95% CI 5.01 - 131.42) between patients 31-35 y and those older than 45 years. No differences were found between patients of 31-35 years and 36-40 years. The OR for total cancellation increased 3.83 (95% CI 2.06 - 7.11) and 19.00 (95% CI 4.56 - 79.11) between women 36-40 years and 41-45 years, and those older than 45 years, respectively. Finally, the oocyte survival rate in patients under 36 years of age was similar to that of our donor program (94% vs. 95%). CONCLUSIONS: Based on this study, we encouraged our patients under than 36 years of age to preserve their fertility for the future.

A healthy HLA-matched baby born by using a combination of aCGH and Karyomapping: the first latin american case.

PGD for HLA typing is a procedure that can be performed when an affected child requires a transplant to treat a non-hereditary disorder related to the hematopoietic and/or immune system. Hematopoietic stem cell transplantation from an HLA-identical donor provides the best treatment option. Three conventional ovarian stimulation procedures for IVF were performed in a couple with a 10-year-old child diagnosed with T-cell acute lymphoblastic leukemia of high risk. Trophectoderm biopsy and aCGH examination were performed on 15 blastocysts, three on the first IVF procedure, four on the second cycle, and eight on the third. Three euploid blastocysts HLA-compatible with the genome of the affected child were identified. One euploid blastocyst HLA-compatible with the affected child was warmed and transferred, resulting in an HLA-matched live birth. In conclusion, combined aCGH for aneuploidy screening and Karyomapping may be performed in a single biopsy procedure.

Mosaic embryo transfer after oocyte in vitro maturation in combination with non-invasive prenatal testing (NIPT)-first report of a euploid live birth.

PURPOSES: The purpose of this study is to describe a healthy life birth after a mosaic embryo transfer in oocyte in vitro maturation (IVM). METHODS: Patient received minimal stimulation, starting on day 3 after menstrual period. No hCG trigger was administered. Oocyte retrieval was performed and oocytes were matured for 30 h. After denuding, mature oocytes were inseminated by ICSI. Embryos were cultured until blastocyst stage and biopsied. RESULTS: One euploid embryo after array comprehensive genome hybridization (aCGH) was diagnostic. However, the next-generation sequencing (NGS) re-analysis showed that embryo was a mosaic for chromosome 13 and 21. Nevertheless, pregnancy ultrasound scans and non-invasive prenatal test (NIPT-Verifi-Illumina) indicated a normal fetus development. Finally, a healthy baby was born after 38 weeks. Its weight was 4480 g, head circumference 36 cm, and total length of 51 cm. To confirm that the baby was chromosomically normal, an NGS test was performed in buccal cells, a normal profile was obtained. CONCLUSIONS: Our finding confirmed that mosaic embryo transfer would bring a healthy offspring.

Desarrollo embrionario preimplantacional, en ciclos de fecundación in vitro con ovodonación y estudio genético / Preimplantational embryo development in in vitro fertilization cycles with oocyte donation and genetic studies

Introducción. Los estudios genéticos preimplantacionales son cada vez más utilizados con la esperanza de conseguir mejores tasas de implantación y nacido vivo, así como una disminución en la tasa de abortos; por ello resulta necesario analizar estos procedimientos. Objetivo. Evaluar el desarrollo preimplantacional in vitro con ovodonación y estudio genético Diseño. Cohortes retrospectivas. Lugar. Laboratorio Pranor, 20082013. Material. Ciclos de fecundación in vitro con ovodonación (FIV-OD). Intervenciones. Se evaluaron 2 077 ciclos de FIV-OD, los cuales fueron clasificados en tres cohortes: 1) ciclos con biopsia embrionaria en día 3, mediante una incisión en la zona pelúcida (ZP) embrionaria, para exéresis de una blastómera (n=527); 2) ciclos con incisión láser en día 4 del desarrollo embrionario, como parte del procedimiento para la biopsia de trofoblasto (n=131); y, 3) ciclos sin intervención (n=1 419). Principales medidas de resultado. Tasa de fecundación, tasa de blastulación. Resultados. No existió diferencia significativa en la tasa de fecundación de las 3 cohortes (75,0%, 74,6% y 75,9%, respectivamente, p=0,31). La tasa de blastulación en la cohorte 1 fue 42,5%, mientras que en la cohorte 3 fue 47% (RR=1,085; IC=1,051 a 1,120; p<0,0001). Adicionalmente, la cohorte 2 tuvo 51,9%, con una diferencia estadísticamente significativa de prevención del riesgo de no blastular con respecto a la cohorte 3 (RR=0,906; IC=0,851 a -0,965; p=0,0017). Conclusiones. El desarrollo preimplantacional hasta blastocisto puede mejorar cuando se utiliza el láser embrionario en día cuatro. Es necesario realizar más estudios para confirmar nuestros resultados.

Introduction: Preimplantational genetic studies are used to achieve a better implantation rate and live birth, as well as to decrease the abortion rate; these techniques should be evaluated. Objective: To evaluate preimplantational embryo development in in vitro fertilization cycles with oocyte donation and genetic studies.. Design: Retrospective cohort study. Setting: Pranor Laboratory, 2008-2013. Material: In vitro fertilization with oocyte donation (IVF-OD) cycles. Interventions: 2 077 cycles of IVF-OD were evaluated, separated in 3 cohorts: Cohort 1, cycles with embryo biopsy on day 3, by means of an incision in the zona pellucida (ZP), for excision of a blastomer (n = 527); Cohort 2, cycles with a laser incision at day 4 of embryo development, for biopsy of the trophoblast in the blastocyst stage (n = 131) and; cohort 3, FIV-OD cycles without any intervention (n = 1 419). Main outcome measures: Fertilization rate, blastulation rate. Results: There was no difference in the fertilization rate among the three groups studied (75.0%, 74.6% and 75.9% respectively, p = 0.31). Blastulation rate in cohort 1, was 42.5%, whereas, in cohort 3, it was 47% (RR = 1.085, CI = 1.0511.120; p <0.0001). In addition, the rate for cohort 2 was 51.9%, with statistical significance, which prevents from non-blastulation risk compared with cohort 3 (RR = 0.906; IC = 0.851-0.965; p = 0.0017). Conclusions: Preimplantational embryo development would improve blastocyst formation when laser is performed on day 4. Further studies are needed to confirm our results.

Maduración in vitro de ovocitos con cultivo extendido hasta estadio de blastocisto: Primer nacimiento reportado en el Perú / In vitro oocyte maturation combined with extended blastocyst embryo culture: Report of the first baby born in Peru

La maduración in vitro de ovocitos (MIV) es una técnica de reproducción asistida muy poco difundida entre los centros de reproducción asistida, debido al bajo éxito en obtener embarazos. Sin embargo, en los últimos años, diferentes estrategias empleadas han demostrado tasas de embarazo similares a las técnicas convencionales de fecundación in vitro (FIV). En el presente reporte, describimos el caso clínico del primer nacido vivo usando MIV en combinación del cultivo extendido hasta estadio de blastocisto.

In vitro oocyte maturation is not yet considered a well-established technique in in vitro fertilization (IVF) laboratories. This is due to a lower pregnancy rates. However in the last few years, reports have shown similar pregnancy rates compared to the conventional IVF techniques. The current report describes the first baby born after an IVM treatment in combination with extended blastocyst culture in Peru.

Primer embarazo con transferencia de embriones seleccionados por morfocinética en el Perú / First pregnancy with transfer of morphokinetics-selected embryos in Peru

La selección embrionaria usando características morfológicas observadas por pocos segundos bajo el microscopio, ha sido la principal herramienta de selección en las técnicas de reproducción asistida. Sin embargo, el desarrollo embrionario es un proceso dinámico que con la introducción de incubadoras con microcámaras integradas, conocidas como incubadoras con sistema Time Lapse, ha permitido registrar eventos morfológicos y cinéticos del desarrollo embrionario que pueden ser útiles como marcadores de selección, denominándolos parámetros 'morfocinéticos'. En este reporte de caso damos a conocer el primer embarazo en el Perú mediante la transferencia de embriones seleccionados por parámetros morfocinéticos en una incubadora con sistema Time Lapse.

Embryo selection by using morphological characteristics has been the main tool to select the best embryo to transfer in assisted reproduction technology (ART). However, embryo development is a dynamic process that cannot be monitored with conventional microscopes. The introduction of incubators with an integrated micro-camera system, denominated time-lapse incubators, has allowed to register morphological and kinetic events in human embryos, be coming useful markers for embryo selection. In this report, we present the first pregnancy in Peru using morphokinetic parameters in a time lapse incubator.

Efficacy of oocyte vitrification combined with blastocyst stage transfer in an egg donation program.

BACKGROUND A successful oocyte vitrification program is important for women with various indications for assisted reproduction technology. The objective of this study was to report the outcome of vitrification of oocytes, obtained through an oocyte donation program, by evaluating the embryo development, pregnancy and implantation rates (IRs) after blastocyst transfer. METHODS A total of 1098 oocytes were obtained from 78 donors. There were 312 oocytes used in the study group (vitrified oocytes) and 786 used in the control group (fresh oocytes). There were 34 recipients who received blastocysts obtained from vitrified oocytes and 58 recipients who received blastocysts from fresh oocytes. The fertilization rate, cleavage rate, embryo quality, pregnancy rate (PR) and IR were compared between groups. RESULTS Vitrified oocytes showed a survival rate of 89.4%. There was no difference in the fertilization rate (76.1 and 87.5%), Day 2 cleavage rate (96.3 and 98.0%) or blastocyst formation rate (41.3 and 45.3%) for the study and control groups, respectively. PRs, IRs and miscarriages rates (MRs) were similar for the study group compared with the control group (PR: 61.8 versus 60.0%; IR: 43.9 versus 42.9%; MR: 9.5 versus 5.9%). CONCLUSIONS The developmental competence of embryos obtained from vitrified oocytes is not affected by the vitrification procedure, since they preserve the potential to be fertilized and to develop into high-quality blastocysts, similar to embryos from fresh oocytes. The successful clinical outcome indicates the use of this procedure for oocyte donation programs and for oocyte storage in general.

In vitro development and pregnancy outcomes for human embryos cultured in either a single medium or in a sequential media system.

OBJECTIVE: To compare a single medium, with medium renewal on day 3, with a sequential media system for development of human embryos to the blastocyst stage and subsequent pregnancy outcome. DESIGN: Prospectively randomized study using donor oocytes. SETTING: Private infertility clinic. PATIENT(S): Oocytes were collected from 47 donors (mean +/- SD age = 25.6 +/- 3.5 years) from March to October 2006 and used for 79 patients (40.6 +/- 5.4 years). The patients were prospectively randomized to have their embryos cultured in a single medium or in a sequential media system. INTERVENTION(S): The donor oocytes were fertilized with fresh patient partner sperm and the resultant zygotes cultured in either a single medium or a sequential media system from day 1 until the blastocyst stage. After endometrial preparation the embryos were transferred to the patients on day 5 or day 6. MAIN OUTCOME MEASURE(S): In vitro embryo developmental characteristics and pregnancy and implantation rates. RESULT(S): In vitro development rates on days 3, 4, and 5 and implantation rates were significantly greater for embryos cultured in the single medium than for those cultured in the sequential media system. CONCLUSION(S): A single medium was as good as or better than a sequential media system for human embryo culture from the zygote to blastocyst stage. The idea that a sequential media system is required for optimal development of the human embryo to the blastocyst stage is highly questionable.

Effect of letrozole at 2.5 mg or 5.0 mg/day on ovarian stimulation with gonadotropins in women undergoing intrauterine insemination.

OBJECTIVE: To determine the effect of combined therapy of letrozole (2.5 mg or 5.0 mg) with recombinant follicle-stimulating hormone (FSH) in comparison with the administration of recombinant FSH alone in an intrauterine insemination (IUI) program. DESIGN: Retrospective study. SETTING: Assisted fertilization program in a specialized infertility center. PATIENT(S): 110 women undergoing IUI and gonadotropin therapy. INTERVENTION(S): Recombinant FSH alone administered from day 3 or combined with letrozole, 2.5 or 5.0 mg/day, on days 3 to 7, and gonadotropins starting on day 7 of the menstrual cycle. Transvaginal ultrasound examinations were done until the dominant follicle reached 18 mm in diameter. Ovulation was triggered with 10,000 IU of human chorionic gonadotropin (hCG), and IUI performed 30 to 40 hours later. MAIN OUTCOME MEASURE(S): Recombinant FSH dose required, number of follicles greater than 14 mm and 18 mm, endometrial thickness, pregnancy rates, miscarriages, and characteristics of newborns. RESULT(S): Women treated with FSH and 5.0 mg/day of letrozole required a lower dose of FSH than the group cotreated with 2.5 mg/day of letrozole or with FSH alone. Throughout most of the follicular phase, the endometrial thickness was statistically significantly less in both letrozole cotreatment groups compared with the FSH control group. By the day of hCG administration, the endometrial thickness was comparable among all the groups. The pregnancy rates were the same with recombinant FSH alone or combined with letrozole. CONCLUSION(S): In terms of cost-effectiveness, 5.0 mg/day of letrozole is more effective than the 2.5 mg/day in cotreatment with no adverse effect on pregnancy rate or outcome.

Sperm chromatin stability and its relationship with fertilization rate after intracytoplasmic sperm injection (ICSI) in an assisted reproduction program.

OBJECTIVE: Evaluate sperm chromatin stability and its relationship with the rate of fertilization after procedures of intracytoplasmic sperm injection (ICSI) in a program of assisted reproduction. DESIGN: Prospective study. SETTING: Institute of Gynecology and Reproduction. PATIENTS: Thirty-three women with their respective partners (12 couples in the study group and 21 couples in the control group) participating in a program of assisted reproduction. The study group was defined as men with >30% of non-decondensed spermatozoa (high sperm chromatin stability). INTERVENTIONS: A part of each seminal sample was used to evaluate sperm chromatin stability under SDS and EDTA treatment and the second aliquot was used for the ICSI procedure. Fertilization was evaluated 16-18 h post sperm injection at a pronuclear stage. The fertilized oocytes were further cultured for 24-48 h before transfer to the patient. MAIN OUTCOME MEASURES: Fertilization rate. RESULTS: Thirty-five oocytes (35.7%) in the group of study and 109 oocytes (78.9%) in the control group showed two pronuclei (P < 0.001). The coefficient of determination between the SDS + EDTA (Grade 2) and rate of fertilization was r (2) = 0.85 (P < 0.001) and the coefficient of regression was 1.72 +/- 0.19 (beta +/- ES) (P < 0.001). CONCLUSIONS: High sperm chromatin stability is a factor which reduces the rate of fertilization after ICSI procedure.