Overcoming male factor infertility with intracytoplasmic sperm injection / Superando o fator masculino de infertilidade com injeção intracitoplasmática de espermatozoides

Summary Objective: To evaluate the effect of male factor infertility on intracytoplasmic sperm injection (ICSI) outcomes compared with a control group presenting isolated tubal factor. Method: This retrospective study included 743 couples undergoing ICSI as a result of isolated male factor and a control group consisting of 179 couples undergoing ICSI as a result of isolated tubal factor, performed in a private university- -affiliated in vitro fertilization center, between January/2010 and December/2016. Patients were divided into two groups according to maternal age: women ≤35 years old and >35 years old. The effects of infertility causes on laboratorial and clinical ICSI outcomes were evaluated using Student's t-test and (2 test. Results: No differences in controlled ovarian stimulation outcomes were observed between male factor cycles and tubal factor cycles in the two age groups. Implantation (male factor 35.5% vs. tubal factor 32.0%, p=0.340), pregnancy (male factor 46.9% vs. tubal factor 40.9%, p=0.184) and miscarriage (male factor 10.3% vs. tubal factor 10.6%, p=0.572) rates were similar between the infertility groups, irrespective of female age. Considering maternal age, the cancelation rate was higher in older women (>35 years old) undergoing ICSI as a result of male factor infertility (17.4% vs. 8.9%, p=0.013). Conclusion: Our results showed that there is no difference in the outcomes of pregnancy between couples with male or tubal factor infertility, which indicates that ICSI surpasses the worse specific outcomes associated with male factor.

Resumo Objetivo: Avaliar o efeito do fator masculino de infertilidade em resultados de injeção intracitoplasmática de espermatozoides (ICSI) em comparação com um grupo controle que apresenta o fator tubário isolado. Método: Este estudo retrospectivo incluiu 743 casais submetidos a ICSI por fator masculino e 179 casais por fator tubário, realizada em um centro privado de fertilização in vitro associado à universidade, entre janeiro de 2010 e dezembro de 2016. Os pacientes foram divididos em dois grupos de acordo com a idade materna: mulheres ≤ 35 e > 35 anos de idade. Os efeitos das causas de infertilidade nos resultados laboratoriais e clínicos da ICSI foram avaliados pelos testes T de Student e Qui-quadrado. Resultados: Não foram observadas diferenças nos parâmetros de estimulação ovariana entre os ciclos com fatores masculinos e com fatores tubários. A taxa de implantação (fator masculino 35,5% vs. fator tubário 32,0%, p=0,340), de gravidez (fator masculino 46,9% vs. fator tubário 40,9%, p=0,184) e de aborto (fator masculino 10,3% vs. fator tubário 10,6%, p=0.572) foram semelhantes entre os grupos de infertilidade, independentemente da idade feminina. Considerando a idade materna, a taxa de cancelamento foi maior em mulheres > 35 anos cuja causa de infertilidade era o fator masculino (17,4% vs. 8,9%, p=0,013). Conclusão: Não há diferenças nos resultados de gravidez entre casais com infertilidade dos fatores masculino ou tubário isolados, o que indica que ICSI supera os piores resultados associados ao fator masculino.

A successful pregnancy using completely immotile but viable frozen-thawed spermatozoa selected by laser / 대한생식의학회지

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

The effects of different types of media on in vitro maturation outcomes of human germinal vesicle oocytes retrieved in intracytoplasmic sperm injection cycles / 대한생식의학회지

OBJECTIVE: Optimizing in vitro maturation (IVM) media to achieve better outcomes has been a matter of interest in recent years. The aim of this prospective clinical trial was to investigate the effects of different media on the IVM outcomes of immature oocytes at the germinal vesicle (GV) stage. METHODS: A total of 400 immature oocytes at the GV stage with normal morphology were retrieved from 320 infertile women aged 31±4.63 years during stimulated intracytoplasmic sperm injection (ICSI) cycles. They were divided into groups of homemade IVM medium (I, n=100), cleavage medium (II, n=100), blastocyst medium (III, n=100), and Sage IVM medium (IV, n=100) and cultured for 24 to 48 hours at 37℃. ICSI was performed, and the rates of fertilization and embryo formation were compared across the four groups. RESULTS: In the 400 retrieved GV oocytes, the total maturation rates showed significant differences in groups I to IV (55%, 53%, 78%, and 68%, respectively, p<0.001). However, there were no significant differences in the fertilization, embryo formation, or arrest rates of metaphase II oocytes across these groups. In all groups, GV maturation was mostly completed after 24 hours, with fewer oocytes requiring 48 hours to mature (p<0.01). Moreover, the rate of high-quality embryos was higher in group IV than in the other groups (p=0.01). CONCLUSION: The quality of the IVM medium was found to affect clinical IVM outcomes. Additionally, blastocyst medium may be a good choice in IVM/ICSI cycles as an alternative IVM medium.

Relationship between meiotic spindles visualization and intracytoplasmic sperm injection outcomes in human oocytes / 대한해부학회지

In assisted reproductive techniques, the operator attempts to select morphologically best embryos to predict embryo viability. Development of polarized light microscope, which evaluates the oocytes' spindles according to birefringence of living cells, had been helpful in oocyte selection. The aim of this study is evaluating the relationship between meiotic spindles visualization and intracytoplasmic sperm injection (ICSI) outcomes in human oocytes. In this study, 264 oocytes from 24 patients with an average age of 30.5±7.5 years with infertility duration of 1 to 10 years were collected. The oocytes were randomly allocated to the control injection group (n=126) and the oocyte imaging group (spindle-aligned group) (n=138). In the spindle-aligned group, the meiotic spindle was identified by means of polarized light microscope to align the spindle at 6 or 12 o'clock. Then the spindle-aligned group was divided into three sub-groups based on spindle morphology: fine, average, and (poor). After ICSI, embryos were checked every 24 hours and scored; 72 hours later, high-grade embryos were transferred intravaginally to uterus. This study showed that the fertilization rate in the spindle-aligned group was higher than the control group (P<0.05). After cleavage, a positive correlation was observed between spindle morphology and embryo morphology. Among the sub-groups of spindle-aligned group, the embryos' morphology of the fine group was better than the other subgroups and embryos of the poor group had lower quality and more fragmentation. The results revealed that the selection of oocytes based on meiotic spindle imaging can significantly improve the rate of fertilization and embryo cleavage and certainly increase the rate of implantation.

Inyección intracitoplasmática de espermatozoides, treinta años después de su implementación / Intracytoplasmic sperm injection, thirty years after its implementation

Resumen: la infertilidad es una enfermedad que se caracteriza por la imposibilidad de lograr un embarazo después de más de 12 meses de relaciones sexuales; como consecuencia, existen tratamientospara manejar este trastorno. Uno de los tratamientos de reproducción asistida es la inyección intracitoplasmática de espermatozoides (ICSI), la cual fue implementada en 1992 para tratar parejas con el factor masculino asociado a la causa de infertilidad. Actualmente, esta técnica es indicada para la infertilidad sin causa aparente, fallas anteriores en los procesos de fertilización in vitro, edad materna avanzada, oocitos de mala calidad, entre otros. La inyección intracitoplasmática de espermatozoides comienza con una estimulación ovárica controlada mediante gonadotropinas y la aspiración folicular para obtener los oocitos. Simultáneamente se procesa la muestra de semen y posteriormente se realiza la microinyección del espermatozoide elegido al interior del oocito. Por otro lado, los parámetros seminales y funcionales han adquirido gran importancia debido al papel determinante en el éxito de la inyección intracitoplasmática de espermatozoides, especialmente la integridad del ADN espermático. Finalmente, la inyección intracitoplasmática de espermatozoides permite que los pacientes con alteraciones seminales tengan la posibilidad de concebir un hijo biológico.En esta revisión se describen los fundamentos de esta técnica y su relación con los parámetros seminales y la fertilidad.

Abstract: Infertility is a disease characterized by the inability to achieve pregnancy after more than 12 months of sexual intercourse; as a result, there are treatments to manage this disorder. One of the treatments of assisted reproduction is the intracytoplasmic sperm injection (ICSI), which was implemented in 1992 to treat couples with male factor associated with the cause of infertility. Currently, this technique is indicated to unexplained infertility, previous in vitro fertilization failures, advanced maternal age, poor quality oocytes, among others. Intracytoplasmic sperm injection begins with a controlled ovarian stimulation and follicular aspiration to obtain oocytes. Simultaneously the semen sample is prepared and the microinjection of the selected spermatozoa into the oocyte is performed. On the other hand, the seminal and functional parameters have become very important due to the determinant role in the success of the intracytoplasmic sperm injection, especially sperm DNA integrity. Finally, intracytoplasmic sperm injection allows patients with seminal alterations have the possibility to conceive a biological child. In this review, the basic of this tecniques and its relationship with sperm parameters and fertility are described.

Fertilization and embryo quality of mature oocytes with specific morphological abnormalities / 대한생식의학회지

OBJECTIVE: To investigate fertilization and embryo quality of dysmorphic mature oocytes with specific morphological abnormalities obtained from intracytoplasmic sperm injection (ICSI). METHODS: The fertilization rate (FR) and embryo quality were compared among 58 dysmorphic and 42 normal form oocytes (control 1) obtained from 35 consecutive ICSI cycles, each of which yielded at least one dysmorphic mature oocyte, performed over a period of 5 years. The FR and embryo quality of 441 normal form oocytes from another 119 ICSI cycles that did not involve dysmorphic oocytes served as control 2. Dysmorphic oocytes were classified as having a dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body (PB). RESULTS: The overall FR was significantly lower in dysmorphic oocytes than in normal form oocytes in both the control 1 and control 2 groups. However, embryo quality in the dysmorphic oocyte group and the normal form oocyte groups at day 3 was similar. The FR and embryo quality were similar in the oocyte groups with a single abnormality and multiple abnormalities. Specific abnormalities related with a higher percentage of top-quality embryos were dark cytoplasm (66.7%), abnormal PB (50%), and cytoplasmic vacuoles (25%). CONCLUSION: The fertilization potential of dysmorphic oocytes in our study was lower, but their subsequent embryonic development and embryo quality was relatively good. We were able to define several specific abnormalities related with good or poor embryo quality.

Increase in concentration of soluble HLA-G in high-quality embryos after intracytoplasmic sperm injection / 대한해부학회지

Non-invasive methods are normally preferred to conventional invasive methods when selecting suitable embryos to improve pregnancy rates after assisted reproduction techniques. One of the most recognized non-invasive methods is to examine the supernatants of embryo culture media. Soluble human leukocyte antigen, class I, G (sHLA-G) antigen is a non-classical class I molecule that has been widely considered as a marker of pregnancy failure or implantation success. In the current study of some Iranian patients, we examined the concentration of sHLA-G at different time points after intracytoplasmic sperm injection and compared the rates to the morphology and quality of the selected embryos. We showed that the concentration of sHLA-G increases over time in high-quality embryos. We conclude that there is a positive relationship between morphology, quality, and sHLA-G concentration. We suggest that this relationship can be used to increase the chance of a successful pregnancy.