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1.
Int. j. morphol ; 37(2): 397-405, June 2019. graf
Article in Spanish | LILACS | ID: biblio-1002234

ABSTRACT

Un embarazo exitoso requiere de una serie de interacciones mediadas por factores hormonales, moleculares y fenómenos de inmunomodulación. Una de estas interacciones es la que ocurre entre el endometrio y el blastocito, previo y durante el proceso de implantación. El objetivo de esta revisión bibliográfica es complementar lo descrito en la literatura clásica de embriología humana sobre interacción de endometrio-blastocito. La búsqueda bibliográfica se realizó en la base de datos MEDLINE usando los términos en inglés "implantation", "endometrium" y "embryo"; además se realizó una búsqueda manual, que incluyó artículos de revistas no indexadas, libros de texto y atlas. Se consideraron criterios de inclusión y exclusión para la selección de los artículos y otros recursos bibliográficos. Entre los criterios de inclusión se consideraron estudios realizados en humanos, artículos de revisión y experimentación, publicados en los últimos 5 años. Como criterios de exclusión se consideraron artículos que utilizaran animales, estudios sobre fertilidad in vitro, patologías asociadas y artículos no relacionados al tema. Una vez completada la selección, se examinaron los textos completos, en los cuales se aplicaron nuevamente los criterios de exclusión. La búsqueda arrojó un total de 560 artículos, cuyo análisis de los títulos y resúmenes resultó en 475 trabajos excluidos, a partir de los diferentes criterios de exclusión antes descritos. Por lo tanto, se obtuvieron 85 artículos, en los cuales se realizó el análisis del texto completo. De estos artículos, se obtuvieron un total de 34 estudios y los contenidos seleccionados en esta revisión fueron: Endometrio, Interacción endometrio trofoblasto, Aposición, Adhesión y Migración-Invasión. Durante la implantación se genera una interacción entre el endometrio y el trofoblasto, con la participación de moléculas reguladoras de proliferación y diferenciación, como factores hormonales, moleculares y de expresión génica. Sin embargo, los mecanismos específicos de acción e interacción deben continuar siendo investigados, para responder interrogantes en el ámbito del crecimiento y desarrollo humano.


A successful pregnancy requires a series of interactions, mediated by hormonal, molecular and immunomodulation phenomena. One of these interactions is between the endometrium and the blastocyst, before and during the implantation process. The objective of this literature review is to complement what is described in the classic human embryology literature on endometrial-blastocyst interaction. The bibliographic search was carried out in the MEDLINE database using the terms "implantation", "endometrium" and "embryo", and a manual search was carried out, which included articles from non-indexed journals, textbooks and atlases. Inclusion and exclusion criteria were considered for the selection of articles and other bibliographic resources, including human studies, review and experimentation articles, published in the last 5 years. Articles with animals as experimental subjects, in vitro fertility studies, associated pathologies and articles not related to the subject were excluded. When the selection was completed, the complete texts were examined, in which the exclusion criteria were applied again The search yielded a total of 560 articles, whose analysis of titles and abstracts resulted in 475 excluded works, in relation to different exclusion criteria described above. Therefore, 85 articles were obtained, in which the complete text analysis was performed. From these articles, a total of 34 studies were obtained and the contents selected in this review were: Endometrium, Endometrium trophoblast, Aposition, Adhesion and Migration-Invasion. During the implantation, aninteraction between the endometrium and the trophoblast is generated, with the participation of regulatory molecules of proliferation and differentiation, such as hormonal, molecular and gene expression factors. However, the specific mechanisms of action and interaction must continue to be investigated, to answer questions in the field of human growth and development.


Subject(s)
Humans , Embryo Implantation , Blastocyst/physiology , Endometrium/physiology , Trophoblasts/physiology
2.
Article in English | WPRIM | ID: wpr-719567

ABSTRACT

BACKGROUND: The standard morphological evaluation has been widely used for embryo selection, but it has limitations. This study aimed to investigate the correlation between morphologic grading and euploidy rate of in vitro fertilization (IVF) preimplantation genetic screening (PGS) and compare the pregnancy rates in young and old ages. METHODS: This is a retrospective study using the medical records of patients who underwent IVF procedures with PGS between January 2016 and February 2017 in a single center. The embryo grades were categorized into 4 groups: excellent, good, fair, and poor. Basic characteristics, euploidy rates, clinical pregnancy (CP) rates and ongoing pregnancy rates were analyzed. RESULTS: The excellent group had significantly higher rate of euploid embryos than fair group (47.82% vs. 29.33%; P = 0.023) and poor group (47.82% vs. 29.60%; P = 0.005). When the four groups were recategorized into two groups (excellent and good vs. fair and poor), they also showed significant difference in euploidy rates (44.52% vs. 29.53%; P = 0.002). When the patients were divided into two groups by age 35, the CP rates for those under and over 35 years old were 44.74% and 47.83%, respectively, which showed no significant difference. CONCLUSION: The significant differences among the euploidy rates of different morphologic embryo grades demonstrated the positive correlations between the morphologic grading of the embryo and the euploidy rate of PGS. Additionally, there was no significant difference between the younger and older patients' CP rates. These findings emphasize the fact that old age patients might benefit from PGS whatever the indication of PGS is.


Subject(s)
Blastocyst , Embryonic Structures , Fertilization in Vitro , Genetic Testing , Humans , In Vitro Techniques , Medical Records , Pregnancy , Pregnancy Rate , Retrospective Studies
3.
Article in English | WPRIM | ID: wpr-758907

ABSTRACT

This study examined the effects of a caffeine treatment to improve nuclear reprogramming in porcine cloned embryos. Embryonic development and the expression of genes related to pluripotency (POU5F1, SOX2, NANOG, and CDX2) were compared after caffeine supplementation during manipulation at different concentrations (0, 1.25, 2.5, and 5.0 mM) and after varying the delayed activation time (control, 1, 2, and 4 h) after fusion. Caffeine added to media during manipulation produced a higher rate of development to blastocysts in the 1.25 mM group than in the other concentration groups (22.8% vs. 16.1%, 16.2%, and 19.2%; p < 0.05). When caffeine was added during the 4 h delayed activation, the 1.25 mM caffeine concentration produced a significantly higher rate of development than those in the other 4 h-activation-delayed caffeine concentration groups (22.4% vs. 9.4%, 14.0%, and 11.1%; p < 0.05). On the other hand, no significant improvement over that in the control group was observed when caffeine was supplemented during both the manipulation period and delayed activation period (16.0% vs. 15.2%), respectively. The levels of POU5F1, SOX2, and NANOG expression in blastocysts were significantly higher in the delayed activation caffeine group (4 h, 1.25 mM) than in the control group (1 h, 0 mM; p < 0.05). In conclusion, a caffeine treatment at 1.25 mM during delayed activation for 4 h can improve the preimplantation development of porcine somatic cell nuclear transfer embryos by activating nuclear reprogramming.


Subject(s)
Blastocyst , Caffeine , Cellular Reprogramming , Clone Cells , Embryonic Development , Embryonic Structures , Female , Hand , Pregnancy
4.
Article in Korean | WPRIM | ID: wpr-760351

ABSTRACT

This study was performed to examine the effects of various macromolecules in in vitro growth (IVG) media on the growth, maturation, and parthenogenesis (PA) of pig oocytes derived from small antral follicles (SAF). Immature oocytes were cultured for two days in IVG medium supplemented with 10% (v/v) fetal bovine serum (FBS), 10% (v/v) pig follicular fluid (PFF), 0.4% (w/v) bovine serum albumin (BSA), or 0.1% (w/v) polyvinyl alcohol (PVA) and then maintained for 44 h for maturation. After IVG, the mean diameters of the SAF treated with FBS, PVA, and no IVG-MAF (113.0–114.8 µm) were significantly larger than that of no IVG-SAF (111.8 µm). The proportion of metaphase II oocytes was higher in PFF (73.6%) than in BSA (43.5%) and PVA (53.7%) but similar to that in the FBS treatment (61.5%). FBS and PFF increased cumulus expansion significantly compared to PVA and BSA while the intraoocyte glutathione content was not influenced by the macromolecules. Blastocyst formation of PA oocytes treated with FBS (51.8%), PFF (50.4%), and PVA (45.2%) was significantly higher than that of the BSA-treated oocytes (20.6%). These results show that the PFF and FBS treatments during IVG improved the growth, maturation, and embryonic development of SAF.


Subject(s)
Blastocyst , Embryonic Development , Female , Follicular Fluid , Glutathione , In Vitro Techniques , Metaphase , Oocytes , Parthenogenesis , Polyvinyl Alcohol , Pregnancy , Serum Albumin, Bovine
5.
Article in English | WPRIM | ID: wpr-785643

ABSTRACT

OBJECTIVE: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts.METHODS: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined.RESULTS: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p<0.05).CONCLUSION: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos.


Subject(s)
Blastocyst , Culture Media , Cumulus Cells , Embryonic Structures , Fertilization , Growth Differentiation Factor 9 , Humans , In Vitro Techniques , Oocytes , Pregnancy , Sperm Injections, Intracytoplasmic
6.
Article in English | WPRIM | ID: wpr-785640

ABSTRACT

OBJECTIVE: We aimed to evaluate the effects of different oxygen conditions (20% [high O₂], 5% [low O₂] and 5% decreased to 2% [dynamic O₂]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture.METHODS: The high-O₂ and low-O₂ groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O₂ group, mouse embryos were cultured from the one-cell to the morula stage under 5% O₂ for 3 days, followed by culture under 2% O₂ to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets.RESULTS: The blastocyst formation rate was significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The total cell number was significantly higher in the dynamic-O₂ group than in the low-O₂ and high-O₂ groups. Additionally, the apoptotic index was significantly lower in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group.CONCLUSION: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.


Subject(s)
Animals , Apoptosis , Blastocyst , Cell Count , Embryo Transfer , Embryonic Structures , Fertilization in Vitro , Humans , In Vitro Techniques , Incubators , Mental Competency , Mice , Morula , Oxygen , Trophoblasts
7.
Article in English | WPRIM | ID: wpr-763348

ABSTRACT

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.


Subject(s)
Aneuploidy , Blastocyst , Chromosome Aberrations , DNA Fragmentation , Embryonic Development , Embryonic Structures , Female , Fertilization in Vitro , Genetic Testing , Germ Cells , Humans , In Vitro Techniques , Maternal Age , Medical Records , Paternal Age , Pregnancy , Pregnancy Rate , Sample Size , Spermatozoa
8.
Article in English | WPRIM | ID: wpr-718516

ABSTRACT

OBJECTIVE: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. METHODS: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. RESULTS: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p < 0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. CONCLUSION: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.


Subject(s)
Blastocyst , Cesarean Section , Embryonic Development , Female , Fertilization , Humans , In Vitro Oocyte Maturation Techniques , Mental Competency , Needles , Oocytes , Pregnancy , Pregnant Women , Sperm Injections, Intracytoplasmic
9.
Article in English | WPRIM | ID: wpr-741724

ABSTRACT

OBJECTIVE: Indications for preimplantation genetic diagnosis (PGD)/preimplantation genetic screening (PGS) cycles and clinical outcomes were evaluated at CHA Gangnam Medical Center. METHODS: This is retrospective cohort study. All patients (n=336) who went through in vitro fertilization (IVF)-PGD/PGS cycles (n=486) between January 2014 and December 2015 were included in Fertility Center of CHA Gangnam Medical Center. Patients underwent IVF-PGD/PGS with 24-chromosome screening. Patients with euploid embryos had transfer of one or 2 embryos in a fresh cycle with any subsequent frozen embryo transfer (ET) cycle. Compared implantation, clinical pregnancy, ongoing pregnancy, and early abortion rates were the main outcome measures. RESULTS: The most common indication for PGD/PGS was recurrent spontaneous abortion (n=160). The chromosome rearrangement cases (n=116) included 24 Robertsonian translocations, 60 reciprocal translocations, 3 inversions, 2 deletions, 4 additions, and 23 mosaicisms. PGS cases rather than the PGD cases showed higher implantation rates (26.4% vs. 20.3%), ongoing pregnancy rates (19.5% vs. 16.4%), and clinical pregnancy rates (28.6% vs. 23.3%). Implantation rates (30.3% vs. 23.7%), clinical pregnancy rates (39.2% vs. 25.2%), and ongoing pregnancy rates (25.7% vs. 17.5%) were significant higher in the blastocyst evaluation group than cleavage stage evaluation group. CONCLUSION: This was the largest study of PGD/PGS for 2 years at a single center in Korea. The pregnancy outcomes of PGD cases are slightly lower than PGS cases. It was confirmed again that success rate of PGD/PGS is higher if biopsy was done at blastocyst than cleavage stage.


Subject(s)
Abortion, Induced , Abortion, Spontaneous , Biopsy , Blastocyst , Cohort Studies , Diagnosis , Embryo Transfer , Embryonic Structures , Female , Fertility , Fertilization in Vitro , Genetic Testing , Humans , Korea , Mass Screening , Outcome Assessment, Health Care , Pregnancy , Pregnancy Outcome , Pregnancy Rate , Preimplantation Diagnosis , Prostaglandins D , Retrospective Studies
10.
Article in English | WPRIM | ID: wpr-716904

ABSTRACT

OBJECTIVE: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. METHODS: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n=300), a group that underwent Cryotop vitrification (group 2, n=300), and a group that underwent our in-house freezing method (group 3, n=300). RESULTS: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p=0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p=0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p=0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable (88.99±10.44, 88.29±14.79, and 86.42±15.23, respectively; p=0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p < 0.001). CONCLUSION: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.


Subject(s)
Animals , Blastocyst , Cell Count , Embryonic Structures , Freezing , Methods , Mice , Survival Rate , Vitrification
11.
Article in English | WPRIM | ID: wpr-713338

ABSTRACT

This study retrospectively assessed whether time-lapse data relating to developmental timing and morphology were associated with clinical outcomes, with the eventual goal of using morphokinetic variables to select embryos prospectively for cryopreservation. In this study, we examined the clinical outcomes of single vitrified-warmed blastocyst transfer cycles that were cultured in a time-lapse incubation system. The morphokinetic variables included uneven pronuclei, an uneven blastomere, multinucleation, and direct, rapid, and irregular division. A total of 164 single vitrified-warmed blastocyst transfer cycles were analyzed (102 cycles of regularly developed blastocysts and 62 cycles of blastocysts with morphokinetic variables). No significant differences in the age of females or the standard blastocyst morphology were found between these two groups. The regularly developed blastocysts showed significantly higher implantation and clinical pregnancy rates than the blastocysts exhibiting morphokinetic variables (30.4% vs. 9.7% and 37.3% vs. 14.5%, respectively; p < 0.01). The blastocysts that exhibited morphokinetic variables showed different mean development times compared with the regularly developed blastocysts. Although morphokinetic variables are known to have fatal impacts on embryonic development, a considerable number of embryos developed to the blastocyst stage. Morphokinetic variables had negative effects on the implantation and clinical pregnancy rates in vitrified-warmed blastocyst transfer cycles. These findings suggest that blastocysts cultured in a time-lapse incubation system should be considered for selective cryopreservation according to morphokinetic variables.


Subject(s)
Blastocyst , Blastomeres , Cryopreservation , Embryo Transfer , Embryonic Development , Embryonic Structures , Female , Humans , Pregnancy , Pregnancy Rate , Prospective Studies , Retrospective Studies , Single Embryo Transfer
12.
Article in Chinese | WPRIM | ID: wpr-689807

ABSTRACT

Various types of medical devices used in assisted reproductive technologies (ART) should be detected for their safety by strict biological assays. Mouse embryo assay(MEA)has been recognized as one of the most important and standardized methods with the threshold more than 80% of blastocyst formation rate (BR) after 96 h culture of fertilized eggs. The disadvantage using BR for embryonic quality control has been concerned as it is ubiquitously dependent of embryonic morphology and the detailed data including molecular and genetic information is obviously missing and incomplete. This leads to the urgent requirement for more sensitive and efficient assessments for the quality control of ART. This study evaluated the reliability of an immunofluorescent MEA by counting total cell and differential number of the cells in the inner cell mass (ICM) and trophectoderm (TE) in the blastocyst. This method improved the traditional MEA, provided a sensitive and powerful platform to assess embryonic developmental viability and should be suggested as a standard assay to be globally used for the quality control of medical devices and pre-clinical procedures in ART.


Subject(s)
Animals , Blastocyst , Embryonic Development , Equipment Safety , Mice , Reproducibility of Results , Reproductive Techniques, Assisted
13.
National Journal of Andrology ; (12): 409-413, 2018.
Article in Chinese | WPRIM | ID: wpr-689742

ABSTRACT

<p><b>Objective</b>Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common genetic renal diseases, which may cause oligoasthenospermia and azoospermia and result in male infertility. This study aimed to analyze the outcomes of preimplantation genetic diagnosis (PGD) in male patients with ADPKD-induced infertility.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data on 7 male patients with ADPKD-induced infertility undergoing PGD from April 2015 to February 2017, including 6 cases of oligoasthenospermia and 1 case of obstructive azoospermia, all with the PKD1 gene heterozygous mutations. Following intracytoplasmic sperm injection (ICSI), we performed blastomere biopsy after 5 or 6 days of embryo culture and subjected the blastomeres to Sureplex whole-genome amplification, followed by haplotype linkage analysis, Sanger sequencing, array-based comparative genomic hybridization to assess the chromosomal ploidy of the unaffected embryos, and identification of the unaffected euploid embryos for transfer.</p><p><b>RESULTS</b>One PGD cycle was completed for each of the 7 patients. Totally, 26 blastocysts were developed, of which 12 were unaffected and diploid. Clinical pregnancies were achieved in 6 cases following 7 cycles of frozen embryo transplantation, which included 5 live births and 1 spontaneous abortion.</p><p><b>CONCLUSIONS</b>For males with ADPKD-induced infertility, PGD may contribute to high rates of clinical pregnancy and live birth and prevent ADPKD in the offspring as well. This finding is also meaningful for the ADPKD patients with normal fertility.</p>


Subject(s)
Abortion, Spontaneous , Genetics , Biopsy , Blastocyst , Comparative Genomic Hybridization , Embryo Transfer , Female , Humans , Infertility, Male , Genetics , Male , Mutation , Polycystic Kidney, Autosomal Dominant , Diagnosis , Genetics , Pregnancy , Pregnancy Outcome , Preimplantation Diagnosis , Retrospective Studies , Sperm Injections, Intracytoplasmic
14.
Chinese Medical Journal ; (24): 1261-1267, 2018.
Article in English | WPRIM | ID: wpr-688133

ABSTRACT

<p><b>Background</b>Despite recent advances that have improved the pregnancy success rates that can be achieved via in vitro fertilization (IVF) therapy, it is not yet clear which blastocyst morphological parameters best predict the outcomes of single blastocyst transfer. In addition, most of the previous studies did not exclude the effect of embryo aneuploidy on blastocysts transfer. Thus, the present study investigated the predictive value of various parameters on the pregnancy outcomes achieved via the transfer of frozen euploid blastocysts.</p><p><b>Methods</b>The study retrospectively analyzed 914 single euploid blastocyst transfer cycles that were performed at the Peking University Third Hospital Reproductive Medical Center between June 2011 and May 2016. The expansion, trophectoderm (TE), and inner cell mass (ICM) quality of the blastocysts were assessed based on blastocyst parameters, and used to differentiate between "excellent", "good", "average", and "poor"-quality embryos. The relationship between these embryo grades and the achieved pregnancy outcomes was then analyzed via the Chi-square and logistic regression tests.</p><p><b>Results</b>For embryo grades of excellent, good, average and poor, the clinical pregnancy rates were 65.0%, 59.3%, 50.3% and 33.3%, respectively; and the live-birth rates were 50.0%, 49.7%, 42.3% and 25.0%, respectively. Both the clinical pregnancy rate (χ = 21.28, P = 0.001) and live-birth rate (χ = 13.50, P < 0.001) increased with the overall blastocyst grade. Both rates were significantly higher after the transfer of a blastocyst that exhibited either an A-grade or B-grade TE, and similarly, an A-grade ICM, than after the transfer of a blastocyst that exhibited a C-grade TE and/or ICM. The degree of blastocyst expansion had no apparent effect on the clinical pregnancy or live-birth rate. All odds ratio were adjusted for patient age, body mass index, length (years) of infertility history, and infertility type.</p><p><b>Conclusions</b>A higher overall euploid blastocyst quality is shown to correlate most strongly with optimal pregnancy outcomes. The study thus supports the use of the described TE and ICM morphological grades to augment current embryo selection criteria.</p>


Subject(s)
Blastocyst , Cell Biology , Physiology , Chi-Square Distribution , Embryo Transfer , Female , Humans , Logistic Models , Odds Ratio , Pregnancy , Pregnancy Outcome , Retrospective Studies
15.
Article in English | WPRIM | ID: wpr-657091

ABSTRACT

Recent advances in stem cell biology have dramatically increased the understanding of molecular and cellular mechanism of pluripotency and cell fate determination. Additionally, pluripotent stem cells (PSCs), including embryonic stem cells and induced pluripotent stem cells, arose as essential resources for disease modeling and cellular therapeutics. Despite these advancements, the epigenetic dysregulation in pluripotency such as the imprinting status, and X chromosome dosage compensation, and its consequences on future utility of PSCs yet remain unresolved. In this review, we will focus on the X chromosome regulation in human PSCs (hPSCs). We will introduce the previous findings in the dosage compensation process on mouse model, and make comparison with those of human systems. Particularly, the X chromosome activation status of human preimplantation embryos, and the regulation of the active X chromosome by human specific lincRNA, X Active Coating Transcript (XACT), will be discussed. We will also discuss the recent findings on higher order X chromosome architecture, and abnormal X chromosome status in hPSCs.


Subject(s)
Animals , Biology , Blastocyst , Chromosomes, Human, X , Compensation and Redress , Embryonic Stem Cells , Epigenomics , Humans , Induced Pluripotent Stem Cells , Mice , Pluripotent Stem Cells , Stem Cells , X Chromosome
16.
Protein & Cell ; (12): 662-674, 2017.
Article in English | WPRIM | ID: wpr-756987

ABSTRACT

Coordination of cell division and cell fate is crucial for the successful development of mammalian early embryos. Aurora kinases are evolutionarily conserved serine/threonine kinases and key regulators of mitosis. Aurora kinase B (AurkB) is ubiquitously expressed while Aurora kinase C (AurkC) is specifically expressed in gametes and preimplantation embryos. We found that increasing AurkC level in one blastomere of the 2-cell embryo accelerated cell division and decreasing AurkC level slowed down mitosis. Changing AurkB level had the opposite effect. The kinase domains of AurkB and AurkC were responsible for their different ability to phosphorylate Histone H3 Serine 10 (H3S10P) and regulate metaphase timing. Using an Oct4-photoactivatable GFP fusion protein (Oct4-paGFP) and fluorescence decay after photoactivation assay, we found that AurkB overexpression reduced Oct4 retention in the nucleus. Finally, we show that blastomeres with higher AurkC level elevated pluripotency gene expression, which were inclined to enter the inner cell mass lineage and subsequently contributed to the embryo proper. Collectively, our results are the first demonstration that the activity of mitotic kinases can influence cell fate decisions in mammalian preimplantation embryos and have important implications to assisted reproduction.


Subject(s)
Animals , Aurora Kinase B , Metabolism , Aurora Kinase C , Metabolism , Blastocyst , Metabolism , Gene Expression Regulation, Developmental , Physiology , Histones , Metabolism , Mice , Phosphorylation , Physiology
17.
Article in English | WPRIM | ID: wpr-219267

ABSTRACT

OBJECTIVE: In vitro culture of preimplantation embryos is improved by grouping embryos together in a drop of media. Individually cultured embryos are deprived of paracrine factors; with this in mind, we investigated whether the addition of a single embryo-secreted factor, interleukin-6 (IL-6), could improve the development of individually cultured embryos. METHODS: Mouse embryos were cultured individually in 2 µL of G1/G2 media in 5% oxygen and supplemented with a range of doses of recombinant mouse or human IL-6. RESULTS: Mouse IL-6 increased hatching at doses of 0.01 and 10 ng/mL compared to the control (93% and 93% vs. 78%, p<0.05) and increased the total number of cells at a dose of 0.1 ng/mL compared to the control (101.95±3.36 vs. 91.31±3.33, p<0.05). In contrast, the highest dose of 100 ng/mL reduced the total number of cells (79.86±3.29, p<0.05). Supplementation with human IL-6 had a different effect, with no change in hatching or total cell numbers, but an increase in the percentage of inner cell mass per embryo at doses of 0.1, 1, and 100 ng/mL compared to the control (22.9%±1.1%, 23.3%±1.1%, and 23.1%±1.1% vs. 19.5%±1.0%, p<0.05). CONCLUSION: These data show that IL-6 improved mouse embryo development when cultured individually in complex media; however, an excess of IL-6 may be detrimental. Additionally, these data indicate that there is some cross-species benefit of human IL-6 for mouse embryos, but possibly through a different mechanism than for mouse IL-6.


Subject(s)
Animals , Blastocyst , Cell Count , Culture Media , Cytokines , Embryology , Embryonic Development , Embryonic Structures , Female , Fertilization in Vitro , Humans , In Vitro Techniques , Interleukin-6 , Mice , Oxygen , Pregnancy
18.
Article in English | WPRIM | ID: wpr-219266

ABSTRACT

OBJECTIVE: Oocyte degeneration often occurs after intracytoplasmic sperm injection (ICSI), and the risk factor is low-quality oocytes. The follicular fluid (FF) provides a crucial microenvironment for oocyte development. We investigated the relationships between the FF volume aspirated from individual follicles and oocyte retrieval, oocyte maturity, oolemma stretchability, fertilization, and development. METHODS: This retrospective study included data obtained from 229 ICSI cycles. Ovarian stimulation was performed according to a gonadotropin-releasing hormone antagonist protocol. Each follicle was individually aspirated and divided into six groups according to FF volume (<1.0, 1.0 to <2.0, 2.0 to <3.0, 3.0 to <4.0, 4.0 to <5.0, and ≥5.0 mL). Oolemma stretchability during ICSI was evaluated using a mechanical stimulus for oolemma penetration, that is, the stretchability was assessed by oolemma penetration with aspiration (high stretchability) or without aspiration (low stretchability). RESULTS: Oocyte retrieval rates were significantly lower in the <1.0 mL group than in the ≥1.0 mL groups (46.0% [86/187] vs. 67.5%–74.3% [172/255 to 124/167], respectively; p<0.01). Low oolemma stretchability was significantly more common in the <1.0 mL group than in the ≥1.0 mL groups during ICSI (22.0% [13/59] vs. 5.8%–9.4% [6/104 to 13/139], respectively; p=0.018). There was a relationship between FF volume and oolemma stretchability. However, there were no significant differences in the rates of fertilization, cleavage, ≥7 cells at day 3, and blastocyst development among all groups. CONCLUSION: FF volume is potentially associated with the stretchability of metaphase II oolemma during ICSI. Regarding oolemma stretchability, ensuring a uniform follicular size during ovarian stimulation is crucial to obtain good-quality oocytes.


Subject(s)
Blastocyst , Clothing , Female , Fertilization , Follicular Fluid , Gonadotropin-Releasing Hormone , Humans , Infertility , Membranes , Metaphase , Oocyte Retrieval , Oocytes , Ovarian Follicle , Ovulation Induction , Retrospective Studies , Risk Factors , Sperm Injections, Intracytoplasmic
19.
IBJ-Iranian Biomedical Journal. 2017; 21 (1): 16-23
in English | IMEMR | ID: emr-185663

ABSTRACT

Backgrund: Imprinted genes are a unique subset of few genes, which have been differentially methylated region [DMR] in a parental origin-dependent manner during gametogenesis, and these genes are highly protected during pre-implantation epigenetic reprogramming. Several studies have shown that the particular vulnerability of imprinting genes during suboptimal pre- and peri-conception micro-environments often is occurred by assisted reproduction techniques [ART]. This study investigated the methylation status of H19/IGF2 DMR at high-quality expanding/expanded human blastocysts donated by healthy individuals to evaluate the risks linked to ART


Method: Methylation levels of H19/IGF2 DMR were analyzed by bisulfite conversion and sequencing at 18 CpG sites [CpGs] located in this region


Result: The overall percentage of methylated CpGs and the proportion of hyper-methylated clones of H19/IGF2 DMR in analyzed blastocysts were 37.85 +/- 4.87% and 43.75 +/- 5.1%, respectively. For validation of our technique, the corresponding methylation levels of peripheral human lymphocytes were defined [49.52 +/- 1.86% and 50%, respectively]


Conclusion: Considering the absence of in vivoproduced human embryos, it is not possible to conclude that the methylation found in H19/IGF2 DMR is actually normal or abnormal. Regarding the possible risks associated with ART, the procedures should be optimized in order to at least reduce some of the epigenetic risks


Subject(s)
Animals, Laboratory , Female , Humans , Male , Blastocyst , Genomic Imprinting , In Vitro Techniques , CpG Islands , Epigenesis, Genetic , Reproductive Techniques, Assisted , Iran
20.
Article in English | WPRIM | ID: wpr-165799

ABSTRACT

OBJECTIVE: The aims of this study were to investigate whether fertilization could induce the resumption of meiosis in mouse oocytes arrested at metaphase I (MI) after in vitro maturation (IVM), and to investigate the effect of Ca²⁺ chelator treatment at the time of fertilization on the transition from MI to metaphase II (MII). METHODS: MII-stage and arrested MI-stage mouse oocytes after IVM were fertilized, and then embryonic development was monitored. Blastocysts from each group were transferred into 2.5 days post-coitum pseudo-pregnant ICR mice. MI oocytes after IVM were treated with a Ca²⁺ chelator to investigate the effect of Ca²⁺ oscillations on their maturation. RESULTS: As insemination time increased, the number of oocytes in the MI group that reached the MII stage also increased. The blastocyst rates and total cell numbers in the MII group were significantly higher than in the MI group. No pregnancy occurred in the MI group, but 10 pregnancies were achieved (10 of 12) in the MII group. The proportion of MI oocytes that matured to MII oocytes after fertilization was significantly higher in the non-treated group than in the Ca²⁺ chelator-treated group. CONCLUSION: The findings that a higher proportion of MI-arrested oocytes progressed to MII after fertilization and that the MI-to-MII transition was blocked by Ca2+ chelator treatments before fertilization indicate that the maturation of MI oocytes to MII oocytes is associated with intracellular Ca²⁺ oscillations driven by fertilization.


Subject(s)
Animals , Blastocyst , Calcium Signaling , Cell Count , Embryonic Development , Female , Fertilization , In Vitro Oocyte Maturation Techniques , In Vitro Techniques , Insemination , Meiosis , Metaphase , Mice , Mice, Inbred ICR , Oocytes , Pregnancy , Spermatozoa
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