Evaluation of physical and neurocognitive development of infants conceived from frozen embryo transfer / 上海交通大学学报（医学版）

Objective • To evaluate the physical and neurocognitive development of infants conceived from frozen embryo transfer (FET). Methods • Two hundred and forty-eight infants (1.5-4 years old) conceived from FET and natural conception (NC) were recruited as the follow-up cohort of FET offspring, and their physical and neurocognitive development were followed up and evaluated. Multiple Logistic regression analysis was used to assess the potential risk of cognitive retardation in FET offspring. Results • There was no significant difference in composition ratio of Z score for height, weight and body mass index between the FET group and the NC control group. Multiple Logistic regression analysis showed that compared with the NC control group, the risk of neurocognitive development abnormalities and retardation was higher in the FET group, especially in fine motor (OR=3.01, 95%CI 1.48-6.11) and social development domains (OR=3.76, 95%CI 1.63-8.69); and in the FET group, the social development risk of female infants was higher than that of male infants. Conclusion • FET may exert a negative impact on the early neurocognitive development of infants.

Correlación entre las concentraciones séricas de hormona antimülleriana y la edad de la mujer con la tasa de recuperación ovular y embarazo / Correlation of the serum levels of Anti-müllerian Hormone and woman age with the rate of ovular recovery and Pregnancy

Resumen OBJETIVO: Evaluar los desenlaces en la recuperación de ovocitos y la tasa de embarazo en pacientes en protocolo de fertilización in vitro de acuerdo con las concentraciones de hormona antimülleriana y la edad de la mujer. MATERIALES Y MÉTODOS: Estudio restrospectivo, transversal, comparativo y observacional efectuado en pacientes en tratamiento de fertilización in vitro en un centro privado de reproducción asistida (enero 2013-enero 2018). La población se dividió en grupo 1 (concentraciones menores de 1 ng/mL de hormona antimülleriana) y grupo B (concentraciones mayores de 1 ng/mL de hormona antimülleriana) para determinar la asociación entre la edad y las concentraciones de hormona antimülleriana y la respuesta a la cantidad de ovocitos recuperados y tasa de embarazo; estos grupos se subdividieron por edades (≤ 29, 30-34, 35-39 y ≥ 40 años). RESULTADOS: Se estudiaron 282 pacientes de las que se eliminaron 35 por expediente incompleto; quedaron 247; 93 en el grupo 1 y 154 en el grupo 2. En el grupo 1 se recuperaron 4.61 vs 7.9 ovocitos (p < 0.001). En las pacientes menores de 29 años con concentraciones de hormona antimülleriana menores de 1 ng/mL la cantidad promedio de ovocitos recuperados fue mayor: 7.8 ovocitos versus los otros grupos de edad con menos de 1 ng/mL. La tasa de embarazo en las pacientes con concentraciones de hormona antimülleriana mayores de 1 ng/mL fue mayor en los grupos de edad menores de 29 años y de 30 a 34 años (p < 0.01). CONCLUSIONES: Independientemente de la edad, cuando las concentraciones de hormona antimülleriana son mayores de 1 ng/mL se logra recuperar mayor cantidad de ovocitos que en quienes tienen menos de esa concentración. Las pacientes menores de 35 años con concentraciones de hormona antimülleriana mayores de 1 ng/mL tienen mayor tasa de embarazo que las de menos de 1 ng/mL. Cuando la edad de la mujer es mayor de 35 años las concentraciones de hormona antimülleriana no influyen en las tasas de embarazo.

Abstract OBJECTIVE: To evaluate the outcomes in oocyte retrieval and the pregnancy rate in patients under in vitro fertilization protocol according to anti-Müllerian hormone concentrations and the age of the woman. MATERIALS AND METHODS: Retrospective, cross-sectional, comparative and observational study carried out in patients undergoing in vitro fertilization treatment in a private assisted reproduction center (January 2013- January 2018). The population was divided into group 1 (lower concentrations of 1 ng / mL of antimülleriana hormone) and group B (concentrations higher than 1 ng / mL of antimülleriana hormone) to determine the association between the age and the antimülleriana hormone concentrations and the answer to the amount of recovered oocytes and pregnancy rate; these groups were subdivided by age (≤ 29, 30-34, 35-39 and ≥ 40 years). RESULTS: We studied 282 patients, of which 35 were eliminated due to incomplete records; there were 247; 93 in group 1 and 154 in group 2. In group 1, 4.61 vs 7.9 oocytes were recovered (p <0.001). In patients younger than 29 years old with antimüllerian hormone concentrations lower than 1 ng / mL, the average number of oocytes retrieved was higher: 7.8 oocytes versus the other age groups with less than 1 ng / mL. The pregnancy rate in patients with antimüllerian hormone concentrations greater than 1 ng / mL was higher in the age groups under 29 years and 30 to 34 years (p <0.01). CONCLUSIONS: Regardless of age, when antimüllerian hormone concentrations are greater than 1 ng/mL, more oocytes are recovered than in those less than that concentration. Patients younger than 35 years old with antimüllerian hormone concentrations lower than 1 ng/mL have a higher pregnancy rate than those less than 1 ng/mL. When the age of the woman is over 35 years old, anti-Müllerian hormone concentrations do not influence pregnancy rates.

Comments on " " from / 中国针灸

In recent years, acupuncture has been used widely as an adjuvant treatment for the in vitro fertilization (IVF). " " published in on May 15, 2018, the research findings do not support the use of acupuncture to improve the rate of live births among the women undergoing IVF. In order to avoid the misunderstanding of the scholars at home and abroad for the clinical effects of acupuncture on IVF assistance, the authors put forward the doubts after the analysis from the following 3 aspects, including the acupuncture scheme, outcomes and result explanation. Additionally, the thinking and suggestions are proposed for the future development of the clinical trials of acupuncture IVF assistance in terms of selecting the proper participants, being abided by the standards of the evidence-based medicine, designing multi-acupuncture schemes and setting up the rational control, as well as conducting the overall analysis of the trial data.

In vitro fertilization (IVF) in mammals: epigenetic and developmental alterations: scientific and bioethical implications for IVF in humans

The advent of in vitro fertilization (IVF) in animals and humans implies an extraordinary change in the environment where the beginning of a new organism takes place. In mammals fertilization occurs in the maternal oviduct, where there are unique conditions for guaranteeing the encounter of the gametes and the first stages of development of the embryo and thus its future. During this period a major epigenetic reprogramming takes place that is crucial for the normal fate of the embryo. This epigenetic reprogramming is very vulnerable to changes in environmental conditions such as the ones implied in IVF, including in vitro culture, nutrition, light, temperature, oxygen tension, embryo-maternal signaling, and the general absence of protection against foreign elements that could affect the stability of this process. The objective of this review is to update the impact of the various conditions inherent in the use of IVF on the epigenetic profile and outcomes of mammalian embryos, including superovulation, IVF technique, embryo culture and manipulation and absence of embryo-maternal signaling. It also covers the possible transgenerational inheritance of the epigenetic alterations associated with assisted reproductive technologies (ART), including its phenotypic consequences as is in the case of the large offspring syndrome (LOS). Finally, the important scientific and bioethical implications of the results found in animals are discussed in terms of the ART in humans.

Resultados preliminares de cultivo intravaginal de ovocitos-inyección intracitoplasmática de espermatozoides (INVO-ICSI), en el centro de fertilidad procrear

Objetivos: Conocer los resultados preliminares de tasa de embarazo por transferencia en las pacientes en quienes se realizó cultivo intravaginal de ovocitos-inyección intracitoplasmática de espermatozoides (INVO-ICSI) en un centro de fertilidad. Diseño: Estudio descriptivo de una serie de casos. Institución: Centro de Fertilidad Procrear, Lima, Perú. Participantes: Parejas con tratamiento de infertilidad. Intervenciones: Se analizó todos los 13 procedimientos de INVO-ICSI realizados en 12 pacientes desde diciembre 2011 a julio 2013. Para el estudio se utilizó el programa SPSS. Principales medidas de resultados: Tasa de gestación. Resultados: La edad promedio de la mujer fue 35,38 años ± 3,35, con promedio de búsqueda de embarazo de 2,25 años ±1,88. La causa de infertilidad en 46,2% era el factor masculino y en 53,2%, el factor femenino. En todos los ciclos se realizó transferencia de embriones. Se obtuvo un total de 5 gestaciones (38,46% por transferencia), de las cuales 3 fueron gestación única (60%) y 2 dobles (40%). Conclusiones: La tasa de embarazo por transferencia mediante el método INVO - ICSI fue comparable y aceptable frente a la de FIV/ICSI reportada en la literatura mundial.

Objectives: To determine preliminary results of pregnancy rate per transfer in patients undergoing intravaginal oocyte culture-intracytoplasmatic sperm injection (INVO-ICSI) at a fertility center. Design: Descriptive, series of cases study. Setting: Procrear Fertility Center, Lima, Peru. Participants: Couples with infertility treatment. Interventions: Thirteen INVO-ICVSI performed in 12 patients from December 2011 through July 2013 were reviewed. SPSS program was used. Main outcome measures: Pregnancy rate. Results: Womens average age was 35.38 years ± 3.35 and were seeking pregnancy 2.25 years ± 1.88 average. Infertility was due to male factor in 46.2% and to female factor in 53.2%. Embryo transfer was done in all cycles and 5 pregnancies were obtained (38.46% of cycles), 3 singleton (60%) and 2 double pregnancy (40%). Conclusions: INVO-ICSI fertilization rate was comparable and acceptable against other fertilization techniques.

Comparación de la calidad embrionaria entre fertilización in vitro (FIV) y cultivo intravaginal de ovocitos (INVO) en el Centro Colombiano de Fertilidad y Esterilidad - CECOLFES, Bogotá, Colombia / Comparing embryo quality between in vitro fertilization (IVF) and intravaginal ovocyte culture (INVO) in the Colombian Centre for Fertility and sterility - CECOLFES, Bogotá, Colombia

Objetivo: comparar la calidad embrionaria y describir las tasas de implantación, embarazo y aborto en las técnicas de fertilización in vitro (FIV) y el cultivo intravaginal de ovocitos. Materiales y métodos: cohortes históricas de pacientes con tratamiento de fertilización in vitro y el cultivo intravaginal de ovocitos en el Centro Colombiano de Fertilidad y Esterilidad (Cecolfes) durante el año 2010. Se incluyeron 137 pacientes aspiradas dentro de los cuatro grupos de estudio: Grupo 1, FIV/Incubadora; Grupo 2, FIV/INVO; Grupo 3, ICSI/INVO, y Grupo 4, ICSI/Incubadora. Se midió el peso de la paciente, el número de ovocitos recuperados y óvulos maduros (MII), la tasa de implantación y la tasa de embarazo y aborto en cada uno de los grupos. Se realizó análisis mediante la prueba de Kruskal-Wallis; la calidad embrionaria fue evaluada con un análisis de covarianza multivariado (MANOVA). Resultados: se observó diferencia significativa en la calidad embrionaria entre las dos técnicas FIV e INVO (p = 0,0388). En la técnica INVO se presentaron mayores tasas de división embrionaria (μ = 7,35/INVO frente a 6,64/Incubadora) y menor fragmentación (μ = 4,67/INVO frente a 4,59/ Incubadora). En cuanto a la tasa de implantación, embarazo y aborto se obtuvieron más altos porcentajes en los grupos INVO. Conclusión: la técnica INVO se asoció a mejor calidad embrionaria. Las tasas de implantación, embarazo y bajas tasas de aborto son semejantes a las descritas en la técnica FIV.

Objective: Comparing embryo quality and describing implantation, pregnancy and abortion rates regarding in vitro fertilization (IVF) and intravaginal oocyte culture (INVO) techniques. Materials and methods: The study involved historical cohorts of patients undergoing IVF and INVO treatment in the Colombian Fertility and Sterility Centre (Centro Colombiano de Fertilidad y Esterilidad – Cecolfes) during 2010. It involved 137 aspirated patients, covering four study groups: Group 1 IVF/incubator, Group 2 IVF/INVO, Group 3 ICSI/INVO and Group 4 ICSI/incubator. The patients’ weight, the number of ovocytes retrieved, mature ovules (M2), implantation rate, pregnancy and abortion rates were measured in each group. The Kruskal-Wallis test was used for statistical analysis; embryo quality was evaluated by multivariate covariance analysis (MANOVA). Results: A significant difference was observed regarding embryo quality between IVF and INVO (p = 0.0388), the INVO technique having higher embryo cleavage rates (μ = 7.35/INVO cf 6.64/ incubator) and lower embryo fragmentation (μ = 4.67/INVO cf 4.59/incubator). INVO groups also had higher percentages concerning their implantation, pregnancy and abortion rates. Conclusion: The INVO technique led to obtaining better quality embryos; implantation, pregnancy and abortion rates were similar to those described for the IVF technique.

Evaluación de la capacidad de desarrollo in vitro de ovocitos bovinos provenientes de vacas con predominancia fenotípica Bos taurus y Bos indicus / Evaluation of in vitro development capacity of bovine oocytes obtained from predominantly Bos taurus and Bos indicus cows

El objetivo de este estudio fue evaluar la capacidad de desarrollo in vitro de ovocitos bovinos de vacas mestizas B. taurus y B. indicus. Los ovocitos fueron recuperados de ovarios de hembras bovinas provenientes de un matadero comercial. Para la obtención de los complejos cumulus-ovocitos (CCO) se realizó la técnica de slicing, seleccionando los ovocitos que tenían al menos una capa de células del cumulus y un citoplasma homogéneo. Los ovocitos seleccionados fueron madurados y fecundados in vitro (MIV-FIV). Se utilizó semen de un toro Brahman puro (B. indicus). Para la evaluación de la MIV y FIV todos los ovocitos se fijaron por al menos 24 h a 4°C en solución metanol-ácido acético (3:1) y teñidos con aceto-orceína al 1,1%. La tasa de maduración de ovocitos de vacas con predominancia fenotípica B. indicus fue del 66,17% mientras que las vacas con predominancia fenotípica B. taurus alcanzaron un 50,94% (P>0,05). En cuanto a la tasa de fecundación se obtuvo un 14,28 y 35,72% de ovocitos penetrados normalmente y anormales, respectivamente, para el grupo de ovocitos con predominancia fenotípica B. indicus. Mientras que para vacas con predominancia fenotípica B. taurus, un 10,22% correspondió a ovocitos penetrados normales y 19,31% de ovocitos penetrados anormales, sin encontrar diferencias estadísticamente significativas en ambos grupos. Los presentes resultados, tanto para la progresión meiótica como para las tasas de fecundación, indican que los ovocitos de vacas mestizas con predominancia fenotípica B. indicus son más competentes en las primeras etapas de desarrollo in vitro que los ovocitos de vacas mestizas con predominancia fenotípica B. taurus.

The aim of this study was to evaluate the in vitro development capacity of bovine oocytes from crossbred B. taurus and B. indicus cows. Oocytes from bovine cows were collected from commercial slaughterhouse. The cumulus-oocyte complex (COC) ovaries were obtained by Slicing technique, selecting those oocytes that had 2 to 3 layers of cumulus cells and homogeneous cytoplasm. After selection oocytes proceed with maturation (IVM) and fertilization in vitro (IVF). It used semen from a pure Brahman bull (B. indicus). For the assessment of IVM as for IVF oocytes were fixed for about 24 hours at 4°C in methanol-acetic acid (3:1) solution and stained with 1.1% aceto-orcein. The maturation rate of oocytes from cows with B. indicus phenotypic predominance was 66.17%, whereas cows with B. taurus phenotypic predominance 50.94% (P>0.05). Fertilization rate obtained in B. indicus phenotypic predominance group was 14.28% of oocytes normal penetrated and abnormal penetrated 35.72%, for cows with a phenotypic predominance B. taurus oocytes normal penetrated were 10.22% and 19.31% of abnormal oocytes penetrated. In conclusion, the present results indicate that oocytes from cows with phenotypic predominance B. indicus are more competent in the early stages of development in vitro than oocytes from cows with phenotypic predominance B. taurus.

A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen

The aim of this study was to compare the effects of spermatozoa separation techniques on sperm quality and in-vitro fertilization (IVF) results for cryopreserved bovine semen. Sephadex, glass wool and Percoll gradient separation techniques were used for sperm separation and sperm motility, morphology and membrane integrity were evaluated before and after separation. Also, cleavage and blastocyst developmental rate were investigated after IVF with sperm recovered by each separation technique. The motility of samples obtained by the three separation techniques were greater compared to the control samples (p < 0.05). The percentage of spermatozoa with intact plasma-membrane integrity, identified by 6-carboxyfluoresceindiacetate/propidium iodide fluorescent staining and the hypo-osmotic swelling test, was highest in the glass wool filtration samples (p < 0.05). The cleavage and blastocyst rate of total oocytes produced from glass wool filtration samples were also higher than the control and Sephadex filtration samples (p < 0.05), but were not significantly different from Percoll separation samples. However, a significantly greater number of cleaved embryos produced by glass wool filtration developed to blastocyst stage than those produced by Percoll separation (p < 0.05). These results indicate that spermatozoa with good quality can be achieved by these three separation techniques and can be used for bovine IVF. In particular, it suggests that glass wool filtration would be the most effective method of the three for improving sperm quality and embryo production for cryopreserved bovine spermatozoa.

The efficacy of low dose GNRH agonist depot in combination with recombinant FSH for ovarian stimulation in in vitro fertilization

Background: Ovarian stimulation is an important process in IVF. Objectives:To assess the efficacy of low dose GnRH agonist depot in combination with recombinant FSH for ovarian stimulation in IVF and evaluate the clinical pregnancy rate of this protocol. Subject and method: A prospective study recruited 60 patients undergoing IVF. The patients was injected intramuscularly with a single dose of diphereline of 1.25 mg on cycle day 21-23. Then recombinant FSH was given subcutaneously on day 3. hCG was injected intramuscularly when at least one follicle reached 18 mm in diameter. Oocytes retrieval in 36 hours and embryo 2 transfer on day after oocytes retrieval. Results: Duration of stimulation was 10.8 \xb1 0.9, average number of oocytes was 3.3 \xb1 4.7, average number of embryos was 6.5 \xb1 4.1, clinical pregnancy rate was 41.7%. Ovarian hyperstimulation rate was 5%, frozen embryo rate was 40 %. Conclusions: The use of low dose GnRH agonist depot in combination with recombinant ISH is effective for ovarian stimulation in IVF.

Effects of Metformin in Patients with Polycystic Ovary Syndrome (PCOS) undergoing In Vitro Fertilization and Embryo Transfer / 대한산부인과학회지

OBJECTIVE: This study was performed to investigate the effects of metformin in PCOS patients undergoing IVF-ET. METHODS: From January 2002 to December 2004, 87 cycles in 32 PCOS patients undergoing IVF-ET at the Infertility clinic of Grace Women's Hospital were randomly divided into two groups and enrolled in this study. The diagnosis of PCOS was made by the criteria from 2003 Rotterdam Consensus. Metformin group (19 patients, 45 cycles) received metformin (Daewoong Pharma Co., Korea; 500 mg three times or two times a day from one or two months before and during IVF cycle) and control group (13 patients, 42 cycles) did not receive metformin. All patients received controlled ovarian hyperstimulation (COH) using gonadotropins (Fostimon, IBSA, Switzerland) with GnRH antagonist (Cetrotide, Serono, Germany). When leading follilces reached 18 mm in diameter, recombinant hCG 250 ?g (OVIDREL, Serono, Italy) was injected. Oocytes were retrieved transvaginally 35 hours later. The luteal phase was supported everyday by progesterone 50 mg IM (Progest, Samil, Korea) with micronized progesterone 200 mg vaginal insertion (Utrogestan, Besins, France). RESULTS: There was no statistical difference in the patients age (34.1+/-3.0 vs 33.5+/-2.9 years), the duration of infertility (4.3+/-1.3 vs 4.5.+/-1.3 years) and BMI (23.5+/-2.4 vs 24.01+/-2.7 kg/m2) among two groups (p> or =0.05 for each). There was also no statistical difference in the indications of IVF among two groups; ovulation factor (27 cycles vs 23 cycles), tubal factor (7 vs 8), male factor (8 vs 7) and other factor (3 vs 4) (p> or =0.05). The duration of ovulation induction in metformin group was significantly shorter when compared with control group (10.1+/-2.3 vs 13.8+/-2.2 days, p<0.05). Total doses of FSH-HP in metformin group were significantly less than that in control group (30.3+/-11.5 vs 39.9+/-11.7 ampules, p<0.05). There was no difference in the number of aspirated oocytes (10.5+/-4.7 vs 9.6+/-3.6), the fertilization rate (65.9+/-17.4% vs 63.2+/-25.9%), implantation rate (30.3+/-11.5% vs 38.1+/-17.3%) and the number of good quality embryo (5.0+/-1.8 vs 4.7+/-2.5, p< or =0.05). Clinical pregnancy rate was higher in metfomin group (33.3% vs. 23.8%), although statistically not significant. CONCLUSION: Although more randomized study is needed, metformin therapy in PCOS patients undergoing IVF has possibility of reducing the duration of COH and the gonadotropin doses.

Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos / 대한불임학회지

OBJECTIVE: In order to acquire the technique for the establishment of human embryonic stem cells (ESC) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. MATERIALS AND METHODS: After F1 hybrid (C57BL x CBA) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). RESULTS: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. CONCLUSION: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.