Well-of-the-well (WOW) versus polyester mesh (PM): a comparison of single-embryo culture systems in bovines

Background: Mexico is innovating in the livestock industry through in vitro generation of bovine embryos with technologies such as well-of-the-well (WOW) and polyester mesh (PM) single-embryo culture systems. These techniques allow to maintain embryos in separate areas of a shared culture medium. Objective: To compare the quantity and quality of bovine embryos produced in WOW and PM culture systems versus the conventional (CG) culture system. Methods: In total, 345 embryos fertilized in vitro were evaluated for blastocyst yield in the three culture systems. To count blastocyst cell numbers, 69 embryos in each system were differentially stained for trophectoderm (TE), inner cell mass (ICM), and apoptotic cells. A qPCR gene expression analysis was performed for embryos in all three systems. Results: The WOW, PM and CG systems developed similar amount of blastocysts (41, 35 and 36%, respectively; p>0.05). Blastocysts in all three systems showed adequate amounts of ICM and apoptotic cells. Blastocysts in the PM system showed a greater number of TE cells [63.7 versus 58.6% in the CG system (p0.05). The ATP5B expression was higher in WOW than in PM (p0.05). The TJP3 expression was higher in PM than in WOW and CG (p<0.05). Expression of ID2 and CLDN4 was higher in WOW than in PM and CG (p<0.05). The biplot graphic from Principal Component Analysis (PCA) revealed that CG was located near degenerated embryos, whereas PM was located near arrested embryos, larger ICM and TE, and TJP3 expression. The WOW was located toward blastocysts, morulae, and expression of CLDN4, ID2 and GNAS. Conclusion: Compared with CG, both the PM and WOW systems are good options for culturing single embryos in the bovine model. Moreover, the PCA results suggest that embryos developed in the WOW system have greater capacity for generating blastocysts with increased ability to form TE and ICM layers, which might improve implantation.

Antecedentes: México está innovando en la industria ganadera a través de la generación in vitro de embriones bovinos con tecnologías de cultivo individual como lo son Pozo dentro de Pozo (WOW) y Malla de Poliéster (PM). Estos mantienen los embriones en áreas separadas mientras comparten un mismo medio de cultivo celular. Objetivo: Comparar la cantidad y calidad de embriones bovinos producidos en los sistemas WOW y PM contra el sistema de cultivo convencional en grupo (CG). Métodos: En total se evaluaron 345 embriones fertilizados in vitro para determinar la producción de blastocistos generados en los tres sistemas. Para contar el número de células por blastocisto, 69 embriones en cada sistema se tiñeron diferencialmente para trofectodermo (TE), masa celular interna (ICM) y células apoptóticas. Se realizó un análisis de expresión génica por qPCR de los embriones obtenidos en los tres sistemas. Resultados: Los sistemas WOW, PM y CG desarrollaron similares cantidades de blastocistos (41, 35 y 36%, respectivamente; p>0,05). Los blastocistos en los tres sistemas mostraron cantidades adecuadas de ICM y células apoptóticas. Los blastocistos en el sistema PM mostraron un mayor número de células TE [63,7% versus 58,6% en el sistema CG (p0,05). La expresión de ATP5B fue mayor en WOW que en PM (p<0,05), pero similar a CG (p<0,05). La expresión de TJP3 fue mayor en PM que en WOW y CG (p<0,05). La expresión de ID2 y CLDN4 fue mayor en WOW que en PM y CG (p<0,05). El gráfico de biplot del análisis de componentes principales reveló que CG se encontró cerca de embriones degenerados, mientras que PM se encontró cerca de embriones en arresto, ICM, TE, y TJP3. El WOW se localizó hacia blastocistos, mórulas y la expresión de CLDN4, ID2 y GNAS. Conclusión: En el modelo bovino los sistemas PM y WOW son buenas opciones para cultivar embriones individuales, ya que se obtienen resultados muy similares a los obtenidos con el sistema CG. Además, los resultados de PCA sugieren que los embriones individuales desarrollados en el sistema WOW generan blastocistos con mayor capacidad de formar TE e ICM, lo que podría mejorar su éxito de implantación.

Antecedentes: O México está inovando na indústria pecuária por meio da geração in vitro de embriões bovinos com tecnologias de cultura de embriões individuais, bem como em poço (WOW) e malha de poliéster (PM). Estes mantêm os embriões em áreas separadas, enquanto compartilham o mesmo meio de cultura de células. Objetivo: Comparar a quantidade e a qualidade de embriões bovinos produzidos nos sistemas de cultura WOW e PM com o sistema convencional de cultura em grupo (CG). Métodos: No total, 345 embriões fertilizados in vitro foram avaliados para determinar a produção de blastocistos gerados nos três sistemas. O número de células por blatocisto foi contado, 69 embriões em cada sistema foram diferencialmente corados para trofectoderme (TE), massa celular interna (ICM) e células apoptóticas. Uma análise de expressão gênica qPCR foi realizada para os embriões obtidos nos três sistemas. Resultados: Os sistemas WOW, PM e CG desenvolveram quantidades semelhantes de blastocistos (41, 35 e 36%, respectivamente; p>0,05). Os blastocistos nos três sistemas mostraram quantidades adequadas de ICM e células apoptóticas. Os blastocistos no sistema PM mostraram um número maior de células TE [63,7 versus 58,6% no sistema CG (p0,05). A expressão de ATP5B foi maior no WOW do que no PM (p<0,05), mas semelhante ao GC (p<0,05). A expressão de TJP3 foi maior no PM do que no WOW e CG (p<0,05). A expressão de ID2 e CLDN4 foi maior no WOW do que no PM e CG (p<0,05). O gráfico biplot da análise de componentes principais revelou que CG foi encontrado próximo a embriões degenerados, enquanto PM foi encontrado próximo a embriões presos, ICM, TE e TJP3. WOW foi encontrado para ter blastocistos, mórulas e a expressão de CLDN4, ID2 e GNAS. Conclusão: Em comparação com o CG, os sistemas PM e WOW são boas opções para a cultura de embriões individuais no modelo bovino. Além disso, os resultados da PCA sugerem que embriões individuais desenvolvidos no sistema WOW têm maior capacidade de desenvolver blastocistos com maior capacidade de formar as camadas TE e ICM, o que poderia melhorar seu sucesso de implantação.

Sperm-specific protein ACTL7A as a biomarker for fertilization outcomes of assisted reproductive technology / 亚洲男科学杂志(英文版)

Obtaining high-quality embryos is one of the key factors to improve the clinical pregnancy rate of assisted reproductive technologies (ART). So far, the clinical evaluation of embryo quality depends on embryo morphology. However, the clinical pregnancy rate is still low. Therefore, new indicators are needed to further improve the evaluation of embryo quality. Several studies have shown that the decrease of sperm-specific protein actin-like 7A (ACTL7A) leaded to low fertilization rate, poor embryo development, and even infertility. The aim of this study was to study whether the different expression levels of ACTL7A on sperm can be used as a biomarker for predicting embryo quality. In this study, excluding the factors of severe female infertility, a total of 281 sperm samples were collected to compare the ACTL7A expression levels of sperms with high and low effective embryo rates and analyze the correlation between protein levels and in-vitro fertilization (IVF) laboratory outcomes. Our results indicated that the ACTL7A levels were significantly reduced in sperm samples presenting poor embryo quality. Furthermore, the protein levels showed a significant correlation with fertilization outcomes of ART. ACTL7A has the potential to be a biomarker for predicting success rate of fertilization and effective embryo and the possibility of embryo arrest. In conclusion, sperm-specific protein ACTL7A has a strong correlation with IVF laboratory outcomes and plays important roles in fertilization and embryo development.

Random sperm DNA fragmentation index is not associated with clinical outcomes in day-3 frozen embryo transfer / 亚洲男科学杂志(英文版)

Damage to sperm DNA was proposed to play an important role in embryonic development. Previous studies focused on outcomes after fresh embryo transfer, whereas this study investigated the influence of sperm DNA fragmentation index (DFI) on laboratory and clinical outcomes after frozen embryo transfer (FET). This retrospective study examined 381 couples using cleavage-stage FET. Sperm used for intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF) underwent density gradient centrifugation and swim up processing. Sperm DFI had a negative correlation with sperm motility (r = -0.640, P < 0.01), sperm concentration (r = -0.289, P < 0.01), and fertilization rate of IVF cycles (r = -0.247, P < 0.01). Sperm DFI examined before and after density gradient centrifugation/swim up processing was markedly decreased after processing (17.1% vs 2.4%, P < 0.01; 65 randomly picked couples). Sperm progressive motility was significantly reduced in high DFI group compared with low DFI group for both IVF and ICSI (IVF: 46.9% ± 12.4% vs 38.5% ± 12.6%, respectively; ICSI: 37.6% ± 14.1% vs 22.3% ± 17.8%, respectively; both P < 0.01). The fertilization rate was significantly lower in high ( ≥25%) DFI group compared with low (<25%) DFI group using IVF (73.3% ± 23.9% vs 53.2% ± 33.6%, respectively; P < 0.01) but was equivalent in high and low DFI groups using ICSI. Embryonic development and clinical outcomes after FET were equivalent for low and high DFI groups using ICSI or IVF. In this study, sperm DFI did not provide sufficient information regarding embryo development or clinical outcomes for infertile couples using FET.

Serum estradiol level on the day of ovulation trigger and pregnancy outcomes in in-vitro fertilisation-intracytoplasmic sperm injection cycles

Background: There is conflicting evidence with regards to the impact of supraphysiologic estradiol levels in in-vitro fertilisation-intracytoplasmic sperm injection (IVF-ICSI) cycles on pregnancy outcomes such as oocyte quality, implantation, and clinical pregnancy. The objective of our study was to evaluate the effect of serum estradiol levels on the day of ovulation trigger on pregnancy outcomes in IVF-ICSI cycles.Methods: We performed a retrospective cohort study, which included eighty-three women who underwent IVF-ICSI and experienced fresh embryo transfer (ET) over one year period. The women included in the study were divided into four groups according to the serum estradiol level on the day of ovulation trigger; Group I: <2000pg/ml, Group II: 2000-3000pg/ml, Group III: 3000-4000pg/ml and Group IV: >4000pg/ml. The outcome measures including number of oocytes retrieved, MII (metaphase II) oocytes, fertilization rate, embryo quality, overall pregnancy rate, implantation rate and clinical pregnancy rate were compared among these four groups.Results: The total number of oocytes, MII oocytes as well as good quality embryos significantly increased from group 1 to group 4. The implantation rate was lowest in group 4 compared to all other groups, although not statistically significant. There was no significant difference in overall and clinical pregnancy rate between the groups.Conclusions: Serum estradiol level shows a positive correlation with the number of oocytes retrieved and good quality embryos. A higher estradiol level does not have a significant negative impact on the implantation rate, overall or clinical pregnancy rate.

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.

Effect of Sperm DNA Fragmentation on Embryo Quality in Normal Responder Women in In Vitro Fertilization and Intracytoplasmic Sperm Injection

PURPOSE: To investigate the associations between sperm DNA fragmentation (SDF) and embryo formation rate in normal responder women to in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). MATERIALS AND METHODS: Fifty-three consecutive, fresh IVF/ICSI cycles performed from 2014 to 2017 were selected. All women were normal responders (4 to 14 mature oocytes were retrieved) and at least one normally fertilized oocyte with two pronuclei was obtained in all cycles. Semen was collected on the day of oocyte retrieval, and SDF levels were measured by sperm chromatin dispersion test (Halosperm assay). At day 3 after insemination, embryo quality was evaluated by morphologic criteria and categorized as A/B/C/D. Top quality embryo were defined as grade A embryos with seven cells or more. RESULTS: SDF levels showed a positive linear correlation with the male's age (r=0.307, p=0.025) and a negative linear correlation with sperm motility (r=−0.491, p70%, the cut-off value SDF was <30.7% for each. Among individuals with SDF <30.7%, the median top-quality or grade A embryo formation rate was significantly higher than that among individuals with SDF ≥30.7% (38.1% vs. 20.0%, p=0.038; 50% vs. 25.0%, p=0.017). CONCLUSION: In normal responder women, high SDF level resulted in low day 3 embryo formation rates. Our results suggest a paternal effect on embryo quality in IVF/ICSI cycles.

Factores clínico-terapéuticos y su relación con la calidad embrionaria en pacientes sometidas a fertilización in vitro / Clinical and therapeutic factors and their relationship with the embryo quality in patients subjected to in vitro fertilization

Introducción: la fertilización in vitro con transferencia embrionaria es el tratamiento de infertilidad más empleado en las técnicas de reproducción asistida. Objetivo: identificar factores clínicos-terapéuticos que inciden en la calidad de los embriones obtenidos por fertilización in vitro, en pacientes infértiles en las que se emplea esta técnica. Métodos: se realizó un estudio longitudinal de corte retrospectivo, en el cual se precisó la influencia de factores clínicos-terapéuticos, tales como, edad materna, dosificación de la hormona folículo estimulante en ciclo previo, número de ovocitos aspirados en el ciclo de fertilización in vitro, análogo de la hormona liberadora de gondadotropinas utilizado y la causa de infertilidad, en la calidad de los embriones cultivados. Resultados: de 419 parejas estudiadas, se obtuvieron tríos embrionarios de buena calidad en el 60,86 por ciento. Hubo diferencias estadísticamente significativas entre las pacientes con buena y mala calidad embrionaria, en cuanto a la edad (p= 0,029), el número de ovocitos aspirados (p< 0,0001) y la causa de infertilidad (p= 0,002). En el análisis multivariado, la causa inexplicable de infertilidad (odds ratio: 0,13 [95 por ciento IC: 0,036-0,49]; p= 0,002) y la edad (odds ratio: 0,96 [95 por ciento IC: 0,81-0,99]; p= 0,042) se asociaron a la abtención de embriones de mala calidad, mientras que el mayor número de ovocitos aspirados se asoció con la obtención de embriones de buena calidad (odds ratio: 1,08 [95 por ciento IC: 1,03-1,13]; p= 0,002). Conclusiones: la obtención de embriones de buena calidad se favorece de un mayor número de ovocitos aspirados; mientras que la infertilidad de causa inexplicable, y el aumento de la edad, la disminuyen(AU)

ntroduction: in vitro fertilization with embryo transfer is the most used infertility treatment in the assisted reproductive techniques. Objective: to identify the clinical and therapeutic factors affecting the quality of embryos obtained from in vitro fertilization in infertile patients undergoing this technique. Methods: a retrospective longitudinal study was conducted to determine the influence of clinical and therapeutic factors such as maternal age, dosage of follicle-stimulating hormone in previous cycle, number of aspired oocytes in the in vitro fertilization cycle, the analogue of the gonadotropin-releasing hormone used and the cause of infertility on the quality of cultured embryos. Results: of 419 studied couples, good quality embryo trios were obtained in 60.86 percent of the sample. There were statistically significant differences between the patients with good and with bad embryo quality in terms of age (p= 0.029), number of aspired ovocytes (p< 0.0001) and cause of infertility (p< 0.002). In multivariate analysis, the unknown cause of infertility (odds ratio: 0.13 [95 percent CI: 0.036-0.49]; p= 0.002) and the age (odds ratio: 0.96 [95 percent CI: 0.81-0.99]; p= 0.042) were associated to obtaining low quality embryos whereas the highest number of aspired ovocytes was associated to good quality embryos (odds ratio: 1.08 [95 percent CI: 1.03-1.13]; p= 0.002). Conclusions: a higher number of aspired oocytes encourages obtaining good quality embryos whereas infertility of unknown cause and older age reduce it(AU)

Repercusiones del índice de masa corporal masculina en los resultados de ICSI / Impact of male body mass index on ICSI outcome

Resumen OBJETIVO: primario, determinar la influencia de la obesidad paterna en la tasa de embarazo, implantación, aborto y de recién nacido en casa en pacientes que reciben tratamientos de fertilización in vitro con ICSI. El objetivo secundario: determinar el efecto del índice de masa corporal del padre en la calidad seminal y embrionaria y en la tasa de fertilización. MATERIALES Y MÉTODOS: estudio retrospectivo, comparativo y observacional al que se incluyeron ciclos ICSI con transferencia embrionaria. Los ciclos se dividieron en 3 grupos según el IMC paterno: peso normal (18.5-24.99 kg/m2), sobrepeso (25-29.99 kg/m2) y obesidad (≥ 30 kg/m2). RESULTADOS: se estudiaron 365 ciclos consecutivos en fresco en los que se encontró un incremento significativo en la tasa de aborto en pacientes con obesidad comparados con el grupo sano (39.0 vs 12.9, p = 0.02), y una significativa disminución en la tasa de recién nacido en casa (38.6 vs 23.6; p = 0.04), además de menor tasa de embriones de excelente calidad, sin afectar la tasa de fertilización con ICSI. CONCLUSIONES: los resultados demuestran que aun cuando las tasas de embarazo e implantación son elevadas en pacientes con sobrepeso y obesidad con técnicas de alta complejidad o ICSI, el desarrollo embrionario, la tasa de aborto y tasa de recién nacido en casa se ven afectados por el incremento del IMC paterno.

Abstract OBJECTIVE: The aim of the present study was primarily to determine the influence of paternal obesity on pregnancy, implantation, abortion and take home baby rates in patients undergoing in vitro fertilization with ICSI. Secondly, to determine the impact of paternal body mass index (BMI) on semen parameters, fertilization rate and embryo quality. MATERIALS AND METHODS: Retrospective, comparative and observational study. 352 cycles of ICSI with embryo transfer were included. They were divided in three groups based on male BMI: Normal (18.5-24.99 kg/m2), overweight (25-29.99 kg/m2), and obesity (≥30 kg/m2). RESULTS: Male obesity was associated with a significant increased miscarriage rate when compared with the normal group (39.0 vs 12.9, p = 0.02) and a significantly lower take home baby rate (23.6 vs 38.6, p = 0.04). We also found a lower rate of top quality embryos, without affecting fertilization with ICSI. CONCLUSION: Our results demonstrate that even though pregnancy and implantation rates are elevated when ICSI is performed in overweight and obese patients, embryo development, miscarriage and take home baby rates are affected with increased paternal BMI.

Effects of Placenta Granules on outcome of mild stimulation protocol / 中成药

AIM To observe the effects of Placenta Granules (Hominis placenta) on endometrial receptivity and embryo outcome.METHODS Ninety infertile patients meeting the kidney Qi deficiency syndrome were randomly divided into H.placenta group (treatment group) and control group.The endometrial thickness,endometrial morphology,blood flow,ovum pick-up number,fertilization number,developmental embryo number,highquality embryo number,pregnancy rate,implantation rate and abnormal pregnancy rate were observed in the two groups.RESULTS There were significant differences in blood flow,numbers of developmental embryo and high-quality embryo (P ＜ 0.05).Ten cases in the control group and fourteen cases in the treatment group reached standard for fresh embryo transplantation.The number of HCG positive in the control group was four cases,which was lower than six cases in the treatment group.There were three cases in the control group on number of clinical pregnancy,and six cases in the treatment group.There were four cases in the control group on implantation rate,and nine cases in the treatment group.All the differences were not statistically significant (P ＞ 0.05).CONCLUSION Placenta Granules can improve the endometrial receptivity and high-quality embryo number in ovarian-stimulation stage under mild stimulation protocol,showing a rising trend in clinical pregnancy rate.

Effects of Placenta Granules on outcome of mild stimulation protocol / 中成药

AIM To observe the effects of Placenta Granules (Hominis placenta) on endometrial receptivity and embryo outcome.METHODS Ninety infertile patients meeting the kidney Qi deficiency syndrome were randomly divided into H.placenta group (treatment group) and control group.The endometrial thickness,endometrial morphology,blood flow,ovum pick-up number,fertilization number,developmental embryo number,highquality embryo number,pregnancy rate,implantation rate and abnormal pregnancy rate were observed in the two groups.RESULTS There were significant differences in blood flow,numbers of developmental embryo and high-quality embryo (P ＜ 0.05).Ten cases in the control group and fourteen cases in the treatment group reached standard for fresh embryo transplantation.The number of HCG positive in the control group was four cases,which was lower than six cases in the treatment group.There were three cases in the control group on number of clinical pregnancy,and six cases in the treatment group.There were four cases in the control group on implantation rate,and nine cases in the treatment group.All the differences were not statistically significant (P ＞ 0.05).CONCLUSION Placenta Granules can improve the endometrial receptivity and high-quality embryo number in ovarian-stimulation stage under mild stimulation protocol,showing a rising trend in clinical pregnancy rate.

Association between Number of Formed Embryos, Embryo Morphology and Clinical Pregnancy Rate after Intracytoplasmic Sperm Injection / Associação entre número de embriões formados, morfologia embrionária e taxa de gravidez clínica após injeção intracitoplasmática de espermatozoides

Abstract Introduction Infertility has a high prevalence in the general population, affecting 5 to 15% of couples in reproductive age. The assisted reproduction techniques ( ART ) include in vitro manipulation of gametes and embryos and are an important treatment indicated to these couples. It is well accepted that the implantation rate is positively influenced by the morphology of transferred embryos. However, we question if, apart from the assessment of embryo morphology, the number of produced embryos per cycle is also related to pregnancy rates in the first fresh transfer cycle. Purpose To evaluate the clinical pregnancy rate according to the number of formed embryos and the transfer of top quality embryos ( TQEs ). Methods In a retrospective cohort study, between January 2011 and December 2012 , we evaluated women who underwent intracytoplasmic sperm injection (ICSI), aged < 40 years, andwith at least 1 formed embryo fresh transferred in cleavagestage. These women were stratified into 3 groups according to the number of formed embryos (1 embryo, 2-3 and ≥ 4 embryos). Each group was divided into 2 subgroups according to the presence or not of at least 1 transferred TQE (1 with TQE; 1 without TQE; 2-3 with TQE, 2-3 without TQE; ≥4with TQE; ≥4withoutTQE). The clinicalpregnancy rateswerecomparedineach subgroup based on the presence or absence of at least one transferred TQE. Results During the study period, 636 women had at least one embryo to be transferred in thefirst fresh cycle (17.8% had 1 formed embryo [32.7% with TQEversus 67.3% without TQE], 42.1% of women had 2-3 formed embryos [55.6% with TQE versus 44.4% without TQE], and 40.1%ofpatientshad ≥4 formedembryos[73.7%withTQEversus26.3%withoutTQE]).The clinical pregnancy rate was significantly higher in the subgroup with ≥4 formed embryos with at least 1 transfered TQE (45.2%) compared with the subgroup without TQE (28.4%). Conclusions Having at least two available embryos and at least one TQE for embryo transfer are predictors of the pregnancy rates.

Resumo Introdução A infertilidade tem uma alta prevalência na população geral, afetando 5 a 15% dos casais em idade reprodutiva. As técnicas de reprodução assistida ( TRA ) incluem a manipulação in vitro de gametas e embriões e são um importante tratamento indicado para esses casais. Sabe-se que a taxa de implantação é positivamente influenciada pela morfologia dos embriões transferidos. No entanto, questiona-se, se além da avaliação da morfologia do embrião, o número de embriões produzidos por ciclo também está relacionado com as taxas de gravidez do primeiro ciclo de transferência fresco. Objetivo Avaliar a taxa de gravidez clínica de acordo com o número de embriões formados e a transferência de embrião com ótima morfologia ( EOM ). Métodos Em um estudo de coorte retrospectivo, entre janeiro de 2011 e dezembro de 2012, avaliamos mulheres submetidas a ICSI com idade < 40 anos e com pelo menos um embrião formado e transferido a fresco em estágio de clivagem. Estas mulheres foram estratificadas em 3 grupos de acordo com o número de embriões formados (1 embrião, 2-3 e ≥ 4 embriões). Cada grupo foi subdividido em 2 subgrupos de acordo com a presença ou não de EOM transferido (1 com EOM; 1 sem EOM; 2-3 com EOM; 2-3 sem EOM; 4 com EOM; ≥4 sem EOM). As taxas de gravidez clínica foram comparadas em cada subgrupo segundo a presença ou não de pelo menos um EOM transferido. Resultados Durante o período do estudo, 636 mulheres tiveram pelo menos 1 embrião para ser transferido no primeiro ciclo a fresco (17,8% possuíram 1 embrião formado [32,7% com EOM versus 67,3% sem EOM], 42,1% das mulheres apresentaram 2-3 embriões formados [55,6% com EOM versus 44,4% sem EOM], e 40,1% das pacientes formaram ≥4 embriões [73,7% com EOM versus 26,3% sem EOM]). A taxa de gravidez clínica foi significativamente maior no subgrupo com ≥4 embriões formados com transferência de pelo menos 1 EOM (45,2%) comparando-se ao subgrupo sem EOM (28,4 % ). Conclusões Ter pelo menos dois embriões e pelo menos um EOM para transferência são fatores preditivos da taxa de gravidez.

WILL THE TYPE OF GNRH ANALOGUE AFFECT THE EMBRYO CLEAVAGE

Aim and Objective:The present study is aimed to carry out the effect of the type of GnRH analogue on embryo cleavage. Materials and Methods:A total of 403 patients who underwent Intra Cytoplasmic sperm injection were included in the study .They were divided in to three groups. Group I- Embryos which cleaved before 27 hours after injection. Group II- Embryos which cleaved after 27 hours, Group III-Embryos which cleavaed before and after 27 hours. The effects of GnRh agonist and antagonist on embryo cleavage were compared between the three groups. Results:All the 403 patients were analysed. There was no difference in the mean age, duration of ovarian stimulation, number of oocytes retrieved, fertilization, cleavage rates and embryo quality between the three groups. Out of 403 patients, early cleavage was observed in 165 patients (40.94 %). Late cleavage was observed in 129 patients (32.01%), both early and late cleavage was observed in 109 patients(27.05%).Out of 227 patients in the agonist protocol the early cleavage was observed in 98 patients (43.17%), late cleavage was observed in 71patients(31.28%),and both early and late cleavage was observed in58 patients(25.55%). Out of 176 patients in the antagonist protocol the early cleavage was observed in 67 patients(38.07%), late cleavage was observed in 58 patients(32.95%),and both early and late cleavage was observed in51 patients(28.98%).P 0.563. We observed there was no statistical significant difference between agonist and antagonist stimulation protocol on embryo cleavage Conclusion: The embryo cleavage was not affected by the type of GnRHanalogue used.

Descarga de la maduración final ovocitaria con gonadotropina coriónica humana o un agonista de la GnRH en donantes de ovocitos: análisis retrospectivo de cohortes

El protocolo de antagonistas de la GnRH (antGnRH) ha permitido el uso del agonista de la GnRH (aGnRH) como inductor de la descarga ovulatoria (DO), en reemplazo de la gonadotropina coriónica humana (hCG). Una ventaja importante con respecto al uso del aGnRH en la DO es que disminuye significativamente el riesgo del síndrome de hiperestimulación ovárica (SHEO), debido a su efecto luteolítico, de importancia en casos de donantes de ovocitos. Objetivos: Comparar la proporción de ovocitos obtenidos, calidad ovocitaria/embrionaria, resultado reproductivo y la incidencia de SHEO en ciclos de donación de ovocitos consecutivos, donde la DO fue con hCG o aGnRH. Adicionalmente, se comparó los ciclos con diagnóstico genético preimplantacional. Diseño: Estudio retrospectivo de cohortes. Institución: Grupo PRANOR, Clínica Concebir, Lima, Perú. Participantes: Donantes y receptoras de ovocitos. Intervenciones: Se utilizó un protocolo antGnRH en 43 donantes de ovocitos para 203 ciclos de estimulación consecutiva y DO con hCG (n=127) o aGnRH (n=76) basados en una decisión clínica y del conteo folicular. Los ovocitos fueron asignados, parte de ellos, a un banco de ovocitos (vitrificación) o a 303 ciclos de receptoras de ovocitos. Principales medidas de resultados: Ovocitos aspirados, tasa de fecundación, calidad embrionaria, formación a blastocisto. Resultados: Se observó diferencias significativas en el número de ovocitos aspirados a favor del grupo aGnRH sobre el grupo hCG (24,09±12,24 versus 18,69±8,93, p=0,002, respectivamente). No hubo diferencias significativas en el resultado global de proporción de ovocitos maduros, tasa de fecundación, calidad embrionaria, formación a blastocisto entre los grupos comparados. La proporción de embriones cromosómicamente normales fueron similares entre los grupos aGnRH (35,2%) y hCG (34,53%), mientras que las tasas globales de embarazo (64,49% versus 62,59%), implantación (45,41% versus 42,38%) y aborto (8,7% versus 8,7%) fueron comparables. La incidencia de SHEO fue reducida significativamente en el grupo de DO con aGnRH (0%), en comparación al grupo hCG (18,9%, p<0,0001). Conclusiones: La DO con el aGnRH es un método seguro y eficaz para ser aplicado en pacientes con alto riesgo de SHEO, como las donantes de ovocitos, eliminando por completo el SHEO. La calidad embrionaria y resultados reproductivos fueron idénticos entre los grupos de DO con aGnRH y hCG.

The GnRH antagonists (GnRHant) protocol has allowed the use of GnRH agonist (GnRHa) as inductor of ovulatory discharge (OD) replacing human chorionic gonadotropin (hCG). An important advantage of GnRHa use in OD is that it significantly decreases the risk of ovarian hyperstimulation syndrome (OHSS) due to its luteolytic effect, important in cases of oocyte donors. Objectives: To compare proportion of oocytes obtained, oocyte/embryo quality, reproductive result and OHSS incidence in consecutive oocyte donation cycles when OD was done with hCG or GnRHa. Cycles with preimplantation genetic diagnosis were compared. Design: Cohort retrospective study. Setting: Grupo PRANOR, Clinica Concebir, Lima, Peru. Participants: Oocyte donors and receptors. Interventions: A GnRHant protocol was used in 43 oocyte donors for 203 consecutive stimulation cycles and OD with hCG (n=127) or GnRHa (n=76) based in clinical decision and follicular count. Oocytes were assigned either to an oocyte bank (vitrification) or to 303 cycles of oocyte receptors. Main outcome measures: Oocytes aspired, fecundation rate, embryo quality, blastocyst formation. Results: There were significant differences in number of oocytes aspired in favor of the GnRHa group with regards to the hCG group (24.09±12.24 versus 18.69±8.93, p=0.002 respectively). There were no significant differences among the groups compared in the global result of mature oocytes proportion, fecundation rate, embryo qulity, blastocyst formation. The proportion of chromosomically normal embryos was similar between GnRHa (35.2%) and hCG (34.53%) groups, and pregnancy (64.49% versus 62.59%), implantation (45.41% versus 42.38%) and abortion (8.7% versus 8.7%) global rates were comparable. OHSS incidence reduced significantly in the OD group with GnRHa (0%) compared with the hCG group (18.9%, p<0.0001). Conclusions: OD with GnRHa is a safe and efficacious method to use in patients with high risk of OHSS such as oocyte donors, as it completely eliminates OHSS. Embryo quality and reproductive results were identical between OD groups with GnRHa and hCG.

Anti-Mullerian Hormone Levels in the Follicular Fluid of the Preovulatory Follicle: A Predictor for Oocyte Fertilization and Quality of Embryo

This prospective study investigated the relationship between anti-Mullerian hormone (AMH) level in the follicular fluid (FF) and the quality of the oocyte and embryo. A total of 65 FF samples from 54 women were included in this study. FF was collected from the largest preovulatory follicle sized> or =20 mm of mean diameter from each ovary. Samples were divided into 3 groups according to the FF AMH levels: below the 33th percentile (low group, FF AMH3.6 ng/mL, n=22). The quality of the ensuing oocytes and embryos was evaluated by fertilization rate and embryo score. FF AMH levels correlated positively with the matched embryo score on day 3 after fertilization (r=0.331, P=0.015). The normal fertilization rate was significantly lower in the low group than in the intermediate group (61.9% vs. 95.5% vs. 77.3%, respectively, P=0.028). Our results suggest that the FF AMH level could be a predictor of the ensuing oocyte and embryo quality.

Improving the probability of pregnancy in endometriosis cases: a study in patients undergoing in vitro fertilization.

Background: Endometriosis is the most common condition underlying infertility in women. To increase the rate of pregnancy in women with endometriosis, embryo selection is performed during in vitro fertilization. This study aims to prove the effectiveness of embryo selection on the rate of pregnancy in a hospital in Jakarta. Methods: This is a cross sectional clinical epidemiology study, performed on endometriosis patients who visited the hospital between 2007 – 2009. Patients were diagnosed with endometriosis using the laparoscopy technique. Embryo selection was performed by assessing the morphology and cell count. Results: We were able to collect data from 72 subjects who underwent IVF during this research period. One subject was dropped out of the program due to immaturity of the oocyte. Successful fertilization was achieved for 65 subjects, but two of them did not undergo embryo transfer. Out of all the subjects undergoing embryo transfer, 26 subjects successfully became pregnant (36.1%). In severe endometriosis cases, pregnancy was achieved with excellent quality embryos (50%) and good-moderate quality embryos (16.7%); but the probability of failure to become pregnant was found to be the same (50%). In mild-moderate endometriosis cases, the probability of pregnancy with excellent quality embryos was 39% compared to 25% chance with good-moderate quality embryos. Regarding the number of embryos that were transferred, we have found that the probability of pregnancy was 50% higher when 3 embryos were transferred, compared to 1 or 2 transferred embryos. Conclusion: This study shows that embryo quality and the number of transferred embryos are relevant to increase the probability of pregnancy in patients undergoing IVF. But because the probability of not achieving pregnancy is not significantly different, we need to find another marker that is more sensitive to assess the quality of embryo transfer.

La fragmentación de ADN espermático, influencia sobre las técnicas de reproducción asistida y la calidad embrionaria / The influence of sperm DNA fragmentation on assisted reproduction techniques and embryo quality

Objetivo: presentar una revisión de la literatura sobre la fragmentación del ADN espermático, su impacto en las técnicas de reproducción asistida y la calidad embrionaria. Metodología: se utilizó la base de datos MEDLINE/PubMed. Los términos de búsqueda fueron: fragmentación espermática, parámetros seminales, técnicas de reproducción asistida y calidad embrionaria. Se revisaron los artículos publicados entre 1999 y 2010. Resultados: los estudios de fragmentación espermática proveen información complementaria acerca de la calidad espermática del paciente, siendo una prueba paralela al análisis seminal. Partiendo del diagnóstico de dichas pruebas se puede tomar la decisión acerca de la técnica de reproducción a emplear. Conclusión: el índice de fragmentación de ADN espermático es una herramienta valiosa para conocer la capacidad fecundante del paciente.

Objective: presenting a review of the literature about sperm DNA fragmentation, its impact on assisted reproduction techniques and embryo quality. Methodology: the MEDLINE/PubMed database was searched; the search terms used were: sperm fragmentation, semen parameters, assisted reproduction techniques and embryo quality. Articles published from 1999 to 2010 were reviewed. Results: sperm fragmentation studies provided complementary information about a patient’s sperm quality, this being a parallel test to semen analysis. Such tests’ diagnoses represent the starting point for taking decisions about the reproduction technique to be used. Conclusion: the sperm ADN fragmentation index represents a valuable tool for ascertaining a patient’s fertilising ability.

Relationship between Pronuclear Scoring and Embryo Quality and Implantation Potential in IVF-ET / 华中科技大学学报（医学）（英德文版）

To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer (WF-ET) program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional WF or intracytoplasmic sperm injec- tion (ICS1). The embryos were classified into groups Z1, Z2, Z3 and Z4. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred. The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4. More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implanta- tion rates of group Z1 were higher (P<0.05) than that of group Z3. These findings suggests that pro- nuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.

Effect of Semen Quality on the Embryo Development / 华中科技大学学报（医学）（英德文版）

To investigate the influences of sperm quality on the zygotes and embryos development,as the role of the paternal factor in early human embryogenesis is gaining more attention because of the application of techniques such as intracytoplasmic sperm injection (ICSI) for the treatment of men infertility, 136 infertility couples with men factors (Group Ⅰ ) were included from May 2002 to January 2004. One hundred and seventy two infertility couples with tube factors (Group Ⅱ ) served as controls. The sperm parameters, gemmates and embryos quality, implantation rate and pregnant rate in both groups were analyzed. It was found that there was no significant differences in the number of oocytes retrieved, the fertilization rate and number of embryos transferred between two groups. Sperm concentration, percentage of motile sperm and percentage of sperm with normal morphology were significantly lower in group Ⅰ than in group Ⅱ (P＜0.01). The proportion of good quality zygotes and good quality embryos were significantly lower in the male infertility group than in the tubal disease group (P＜0.05). Implantation rate and pregnancy rate were similar in two groups. It was concluded that spermatozoa is involved in the embryo quality, even in the early stages of development, which limited the treatment potency of IVF procedure.

Recombinant Human Follicle Stimulating Hormone (rFSH) versus Highly Purified Urinary FSH (uFSH): Oocyte and Embryo Quality / 대한산부인과학회잡지

OBJECTIVE: To estimate the efficacy of recombinant human follicle stimulating hormone (rFSH) versus highly purified urinary human FSH (uFSH) in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization and embryo transfer (IVF-ET). METHODS: From 1 January 2001 to 31 August 2001, A total of 254 cycles from 241 patients who attended infertility clinic at Samsung cheil hospital was enrolled in this study. With pituitary down regulation using GnRH agonist by short protocol, rFSH (Puregon(R), Organon, Netherlands) was administered in 131 cycles and uFSH (Metrodin-HP(R), Serono, Switzerland) was administered in 123 cycles. We analyzed ovarian response, pregnancy rate, live birth rate, oocyte quality and embryo quality. RESULTS: The clinical characteristics of two groups were not different. Total FSH dosages (1322.3+/-526.2 IU versus 2124.4+/-881.9 IU, p<0.001) and dosages per retrieved oocyte (90.6+/-36.0 IU versus 138.0+/-57.2 IU, p<0.001) were significantly lower in rFSH group than uFSH group. Clinical pregnancy rate and live birth rate of two groups were not significantly different. The rate of good quality oocyte (Grade I and II) from retrieved oocytes was higher in rFSH group (68.2% versus 64.8%, p=0.024), but after preincubating oocytes for 4 to 6 hours and removing cumulus cells in intracytoplasmic sperm injection (ICSI) cycles, nuclear maturity of oocytes were not significantly different. The quality of transferred embryos were not significantly different too. CONCLUSION: rFSH offered more effective ovarian response in COH and better quality of retrieved oocytes, compared with uFSH.

The Impact of Oocyte Maturity and Embryo Quality on Pregnancy Rate of In Vitro fertilization-EmbryoTransfer(IVF-ET) Program / 대한산부인과학회잡지

There are a number of factors affecting the pregnancy rate in in vitro fertilization(IVF): the etiology of infertility, ages of husband and wife, the type of ovarian stimulation,the number of aspirated oocytes, the maturity of oocyte, embryo quality, the number offertilized oocytes, the cumulative embryo score, and the number of embryos transferred. Amongthem the maturity of oocyte and the quality of embryo appear to be the most importantfactors influencing the pregnancy rate. The aim of this study designed to evaluate the effectof oocyte maturity, embryo quality, the number of embryos transferred, and the cumulativeembryo score to fertilization rate on whether the patient conceived or not. A total of146 consecutive cycles of in vitro fertilization-embryo transfer(IVF-ET) were studied usingtwo ovarian hyperstimulation protocols; follicular stimulating hormone(FSH)/ humanmenopausal gonadotropin(HMG)/human chorionic gonadotropin(hCG) and gonadotropin releasinghormone agonist(GnRH-a)/FSH/HMG/hCG.Following were the results.1. The quality and quantity of oocytes and embryos, fertilization rate, and cumulativeembryo score between two ovarian hyperstimulation protocols were not different.2. The more mature oocytes exhibited, the greater fertilization rates in both methods(p < 0.01).3. The average numbers of embryos transferred were 3.65 +/- 0.27 in conceived groupand 2.54 +/- 0.18 in nonconceived group, which was statistically higher in conceived group(p < 0.001).4. The pregnancy rate was higher if any matured embryo(Grade I) was present amongthe embryos transferred(p < 0.05).5. The average cumulative embryo score was 69.70 +/- 6.13 inconceived group and 43.51 +/- 3.41 in nonconceived group, which was statistically higher in conceived group(p < 0.001).These data suggest that the pregnancy rate is higher when the oocyte maturation isBetter, the number of embryos transferred is greater, there is at least one matured embryoamong the embryos transferred, and the cumulative embryo score is higher.