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1.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 38(9): 845-848, 2021 Sep 10.
Artigo em Chinês | MEDLINE | ID: mdl-34487527

RESUMO

OBJECTIVE: To assess the application value of mapping allele with resolved carrier status (MaReCs) technique for preimplantation genetic testing (PGT). METHODS: The characteristics of MaReCs for PGT and outcome of patients were retrospectively analyzed. RESULTS: Compared with those who could not use the technique, carriers who have used the MaReCs technique were younger, had significantly higher level of anti-Mullerian hormone, more antral follicles, occytes, mature occytes, biopsied embryos and euploid embryos, and lower risks for de novo chromosomal abnormality (P<0.05). It was necessary for couples with fewer oocytes, mature oocytes and balstocyst to preserve discarded embryos to facilitate the test. Carriers who have used the MaReCs technique had higher clinical pregnancy rate and abortion rate compared with those undergoing routine PGT, albeit no significant difference was found between the two groups (P> 0.05). Carriers undergoing MaReCs test could preferentially select embryos with normal chromosome structures for the transfer. CONCLUSION: Application of MaReCs has a prerequisite for having a minimum number of occytes and biopsied embryos and using discarded embryos sometimes. MaReCs is efficient for the detection of carrier status of embryos and attaining higher rate of pregnancy and live birth, which can significantly improve the outcome for couples carrying chromosomal translocations.


Assuntos
Diagnóstico Pré-Implantação , Translocação Genética , Alelos , Aneuploidia , Blastocisto , Feminino , Fertilização In Vitro , Testes Genéticos , Humanos , Gravidez , Estudos Retrospectivos
2.
Int J Gynaecol Obstet ; 155(1): 13-15, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34520054

RESUMO

FIGO (the International Federation of Gynecology and Obstetrics) supports assisted reproductive technologies (ART) to achieve pregnancy and supports their availability in all nations. However, the increased frequency of preterm birth must be taken into account. Therefore, before in vitro fertilization (IVF) is started, other approaches, including expectant management, should be considered. Single embryo transfer is the best approach to ensure a live, healthy child. However, increased risks for preterm birth are also associated with a singleton IVF pregnancy and should be discussed and contrasted with spontaneous conception. Increased preterm birth and other adverse pregnancy outcomes in singleton IVF cycles warrant investigations to elucidate and mitigate. Minimizing embryo manipulation during cell culture is recommended. Increased risk of preterm birth and other pregnancy complications in ART could reflect the underlying reasons for infertility. This information should be discussed and further explored.


Assuntos
Nascimento Prematuro , Feminino , Fertilização , Fertilização In Vitro , Humanos , Recém-Nascido , Gravidez , Resultado da Gravidez , Nascimento Prematuro/epidemiologia , Técnicas de Reprodução Assistida , Estudos Retrospectivos
3.
Nutrients ; 13(7)2021 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-34371958

RESUMO

Female reproductive aging is an irreversible process associated with a decrease in oocyte quality, which is a limiting factor for fertility. Previous studies have shown that dehydroepiandrosterone (DHEA) has been shown to improve in vitro fertilization (IVF) outcomes in older women. Herein, we showed that the decline in oocyte quality with age is accompanied by a significant decrease in the level of bioenergetic metabolism genes. We compared the clinical characteristics between groups of infertile women who either received DHEA or did not. Treatment with DHEA may enhance oocyte quality by improving energy production and metabolic reprogramming in cumulus cells (CCs) of aging women. Our results showed that compared with the group without DHEA, the group with DHEA produced a large number of day-three (D3) embryos, top-quality D3 embryos, and had improved ongoing pregnancy rate and clinical pregnancy rate. This may be because DHEA enhances the transport of oxidative phosphorylation and increases mitochondrial oxygen consumption in CCs, converting anaerobic to aerobic metabolism commonly used by aging cells to delay oocyte aging. In conclusion, our results suggest that the benefit of DHEA supplementation on IVF outcomes in aging cells is significant and that this effect may be mediated in part through the reprogramming of metabolic pathways and conversion of anaerobic to aerobic respiration.


Assuntos
Desidroepiandrosterona/administração & dosagem , Suplementos Nutricionais , Metabolismo Energético , Infertilidade Feminina/metabolismo , Oócitos/metabolismo , Biogênese de Organelas , Adulto , Envelhecimento , Senescência Celular , Células do Cúmulo/metabolismo , Feminino , Fertilização In Vitro , Humanos , Mitocôndrias/metabolismo , Fosforilação Oxidativa , Consumo de Oxigênio , Gravidez , Resultado da Gravidez
6.
Theriogenology ; 173: 144-155, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34390905

RESUMO

The beneficial effect of glutathione (GSH) on the in vitro maturation (IVM) of bovine/porcine oocytes has been confirmed; however, the antioxidant effect of exogenous GSH supplementation on the IVM of ovine oocytes has not been determined. In this study, ovine cumulus oocyte complexes (COCs) were classified into three groups according to the layer number of cumulus cells (the Grade A group has more than five layers, the Grade B group has three to four layers and the Grade C group has less than three layers). After in vitro culture of COCs in the presence of exogenous GSH, the meiotic competence of ovine oocytes was assessed by analyzing nuclear maturation to metaphase II (MII) stage, cortical granules (CGs) dynamics, astacin like metalloendopeptidase (ASTL) distribution, histone methylation pattern, reactive oxygen species (ROS) production, mitochondrial activities and genes expression. After in vitro fertilization (IVF), assessments of embryonic development were conducted to confirm the effects of exogenous GSH supplementation. The results showed that exogenous GSH not only enhanced the maturation rates of the Grade B and Grade C groups but also promoted CGs dynamics and ASTL distribution of the Grade A, B and C groups (p < 0.05). Exogenous GSH increased the mitochondrial activities of the Grade A, B and C groups and decreased the ROS production levels of oocytes (p < 0.05), regardless of the layer number of cumulus cells. Moreover, exogenous GSH promoted the expression levels of genes related with oocyte maturation, antioxidant activity and antiapoptotic effects in the Grade B and Grade C groups (p < 0.05). The expression levels of H3K4me3 and H3K9me3 in the Grade B and Grade C groups were promoted after exogenous GSH supplementation (p < 0.05), consistent with the expression levels of genes related with histone methylation (p < 0.05). In addition, exogenous GSH strongly promoted the embryonic developmental competence of Grade B and Grade C groups (p < 0.05). Taken together, our findings provide foundational evidence for the free radical scavenging potential of exogenous GSH in the in vitro developmental competence of ovine oocytes, especially oocytes from COCs lacking cumulus cells. These findings, which demonstrated the potential for improving the quality of ovine oocytes during IVM, will contribute to researches on GSH applications and the efficiency of assisted reproductive technology for ovine breeding.


Assuntos
Glutationa , Técnicas de Maturação in Vitro de Oócitos , Animais , Células do Cúmulo , Suplementos Nutricionais , Desenvolvimento Embrionário , Feminino , Fertilização In Vitro/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos , Gravidez , Ovinos
7.
Eur J Obstet Gynecol Reprod Biol ; 264: 266-270, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34340097

RESUMO

OBJECTIVE: The aim of this retrospective case-control study was to analyze the effect of administering indomethacin after triggering final oocyte maturation in patients with poor ovarian response (POR) on the cycle cancellation rate due to premature ovulation (PO), the number of oocytes retrieved and the clinical outcomes of IVF cycles. STUDY DESIGN: A total of 214 patients with POR, diagnosed according to the Bologna criteria, who underwent fresh IVF cycle via flexible gonadotrophin-releasing hormone antagonist (GnRH-ant) protocol were enrolled in the study. The control group consisted of 100 patients, whereas the indomethacin group included 114 patients who received 100 mg rectal indomethacin administered twice within the same day (twelve hours apart) -starting at twelve hours after triggering. Cycle cancelation rates (CCR), number of oocytes retrieved (nOR), implantation rates (IR), biochemical pregnancy (BP) and clinical pregnancy loss rates (CPL), ongoing pregnancy rates (OPR) and live birth rates (LBR) were compared between the indomethacin and control groups. RESULTS: The CCR rate was significantly lower in the indomethacin group (1.8%) compared to the control group (1.8% vs %12%, p = 0.01). In the control group, those with cycle cancellation were older than those without cycle cancellation (mean age 42.2 ± 2.3 years vs. 39.36 ± 4.3 years, p = 0.001) and had lower anti-Müllerian hormone levels and lower antral follicle count (0.59 ± 0.2 ng/mL vs 0.79 ± 0.2 ng/mL, p = 0.001 and 4 ± 0.6 vs 5.7 ± 1.7, p = 0.001, respectively). In multivariable analysis, when the dependent variable in the logistic regression model was coded as the absence of cycle cancellation, it was observed that only indomethacin had a statistically significant effect on cycle cancellation (ß = -1.931, standard error = 0.832, Exp(B) = 0.145, p = 0.020). nOR was higher in the indomethacin group than control group but the difference did not reach significance (p = 0.07). Moreover, the IR, OPR and LBR, BP and CPL values were similar in the indomethacin and control groups (p > 0.05). CONCLUSIONS: Based on data from this study, it can be concluded that indomethacin reduces cycle cancelation due to PO in patients with POR -without compromising implantation and pregnancy rates. However, further randomized controlled trials with larger sample sizes are required to clarify the definitive effect of indomethacin in the treatment of patients with POR.


Assuntos
Fertilização In Vitro , Indução da Ovulação , Adulto , Estudos de Casos e Controles , Feminino , Hormônio Liberador de Gonadotropina , Humanos , Indometacina/uso terapêutico , Oócitos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
10.
Hum Reprod ; 36(9): 2506-2513, 2021 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-34364311

RESUMO

STUDY QUESTION: Does the immune response to coronavirus disease 2019 (COVID-19) infection or the BNT162b2 mRNA vaccine involve the ovarian follicle, and does it affect its function? SUMMARY ANSWER: We were able to demonstrate anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgG in follicular fluid (FF) from both infected and vaccinated IVF patients, with no evidence for compromised follicular function. WHAT IS KNOWN ALREADY: No research data are available yet. STUDY DESIGN, SIZE, DURATION: This is a cohort study, composed of 32 consecutive IVF patients, either infected with COVID-19, vaccinated or non-exposed, conducted between 1 February and 10 March 2021 in a single university hospital-based IVF clinic. PARTICIPANTS/MATERIALS, SETTING, METHODS: A consecutive sample of female consenting patients undergoing oocyte retrieval was recruited and assigned to one of the three study groups: recovering from confirmed COVID-19 (n = 9); vaccinated (n = 9); and uninfected, non-vaccinated controls (n = 14). Serum and FF samples were taken and analyzed for anti-COVID IgG as well as estrogen, progesterone and heparan sulfate proteoglycan 2 concentration, as well as the number and maturity of aspirated oocytes and day of trigger estrogen and progesterone measurements. Main outcome measures were follicular function, including steroidogenesis, follicular response to the LH/hCG trigger, and oocyte quality biomarkers. MAIN RESULTS AND THE ROLE OF CHANCE: Both COVID-19 and the vaccine elicited anti-COVID IgG antibodies that were detected in the FF at levels proportional to the IgG serum concentration. No differences between the three groups were detected in any of the surrogate parameters for ovarian follicle quality. LIMITATIONS, REASONS FOR CAUTION: This is a small study, comprising a mixed fertile and infertile population, and its conclusions should be supported and validated by larger studies. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study to examine the impact of SARS-Cov-2 infection and COVID-19 vaccination on ovarian function and these early findings suggest no measurable detrimental effect on function of the ovarian follicle. STUDY FUNDING/COMPETING INTEREST(S): The study was funded out of an internal budget. There are no conflicts of interest for any of the authors. TRIAL REGISTRATION NUMBER: CinicalTrials.gov registry number NCT04822012.


Assuntos
COVID-19 , Folículo Ovariano , SARS-CoV-2 , Vacinas contra COVID-19 , Estudos de Coortes , Feminino , Fertilização In Vitro , Humanos , Folículo Ovariano/fisiopatologia , RNA Mensageiro , Vacinação
11.
BMJ Open ; 11(8): e047974, 2021 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-34408042

RESUMO

INTRODUCTION: Many patients demonstrate an insufficient endogenous luteinising hormone (LH) concentration during ovarian stimulation. With traditional fixed or flexible gonadotropin-releasing hormone (GnRH) antagonist protocols, antagonist administration may further reduce LH activity. Previously, we proved that LH can be used as an indicator for the timing and dosage of antagonist. Patients with a persistently low LH concentration during ovarian stimulation may not require antagonists, whereas antagonist administration can affect reproductive outcomes. To further explore this hypothesis, we designed a randomised clinical trial to compare the LH-based flexible GnRH antagonist protocol with traditional flexible GnRH antagonist protocol in women with normal ovarian response. METHODS AND ANALYSIS: This study was a multicentre, parallel, prospective, randomised, non-inferiority study. The primary efficacy endpoint was cumulative ongoing pregnancy rate per cycle. The study aimed to prove the non-inferiority of cumulative ongoing pregnancy rate per cycle with an LH-based flexible GnRH antagonist protocol versus traditional flexible GnRH antagonist protocol. Secondary endpoints were the high-quality embryo rate, clinical pregnancy rate and cancellation rate. Differences in cost-effectiveness and adverse events were evaluated. The cumulative ongoing pregnancy rate per cycle in women with normal ovarian response was 70%. Considering that a non-inferiority threshold should retain 80% of the clinical effect of a control treatment, a minimal clinical difference of 14% (one-sided: α, 2.5%; ß, 20%) and a total of 338 patients were needed. Anticipating a 10% drop-out rate, the total number of patients required was 372. ETHICS AND DISSEMINATION: This trial has been approved by the Institutional Ethical Committee of Beijing Chao-Yang hospital. All participants in the trial will provide written informed consent. The study will be conducted according to the principles outlined in the Declaration of Helsinki and its amendments. Results of this study will be disseminated in peer-reviewed scientific journals. TRIAL REGISTRATION NUMBER: ChiCTR1800018077.


Assuntos
Fertilização In Vitro , Hormônio Liberador de Gonadotropina , Feminino , Humanos , Hormônio Luteinizante , Estudos Multicêntricos como Assunto , Indução da Ovulação , Gravidez , Estudos Prospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto
12.
Theriogenology ; 173: 93-101, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34365139

RESUMO

Cryopreservation of both gametes and embryos, both for storage and for the preservation of their developmental capacity is a critical aspect of assisted reproductive technology. The survival of reproductive material following cryopreservation protocols is not only vital to clinical applications in the human in vitro fertilisation clinic, but is also important in the in vitro production of livestock embryos. The ability to routinely cryopreserve oocytes and embryos of livestock species has the potential to improve animal welfare, reduce environmental impact, and reduce the associated costs for breeding companies through the reduction of live animal transportation. Unfortunately, frozen oocytes and embryos are regularly documented to contain a higher proportion of apoptotic cells compared to their non-frozen counterparts, with freezing procedures thought to trigger apoptotic pathways of cell death. Comparisons between frozen and non-frozen samples also show changes in the gene expression of apoptotic factors such as Bcl-2 and Bax in response to cryopreservation. Apoptotic inhibition has the potential to improve cryosurvival, and how to achieve this is subject to debate. Here, we review how exposure to low temperatures during cryopreservation may be responsible for the abnormal activation of apoptotic pathways in mammalian oocytes and embryos, and discuss the ways in which they can be influenced to improve cryopreservation protocols, particularly in agriculturally important species.


Assuntos
Criopreservação , Embrião de Mamíferos , Animais , Apoptose , Criopreservação/veterinária , Fertilização In Vitro/veterinária , Oócitos
13.
Int J Mol Sci ; 22(16)2021 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-34445470

RESUMO

In regular IVF, a portion of oocytes exhibit abnormal numbers of pronuclei (PN) that is considered as abnormal fertilization, and they are routinely discarded. However, it is known that abnormal ploidy still does not completely abandon embryo development and implantation. To explore the potential of cytoplasm from those abnormally fertilized oocytes, we developed a novel technique for the transfer of large cytoplasm between pronuclear-stage mouse embryos, and assessed its impact. A large volume of cytoplast could be efficiently transferred in the PN stage using a novel two-step method of pronuclear-stage cytoplasmic transfer (PNCT). PNCT revealed the difference in the cytoplasmic function among abnormally fertilized embryos where the cytoplasm of 3PN was developmentally more competent than 1PN, and the supplementing of fresh 3PN cytoplasm restored the impaired developmental potential of postovulatory "aged" oocytes. PNCT-derived embryos harbored significantly higher mitochondrial DNA copies, ATP content, oxygen consumption rate, and total cells. The difference in cytoplasmic function between 3PN and 1PN mouse oocytes probably attributed to the proper activation via sperm and may impact subsequent epigenetic events. These results imply that PNCT may serve as a potential alternative treatment to whole egg donation for patients with age-related recurrent IVF failure.


Assuntos
Núcleo Celular/patologia , Citoplasma/patologia , Embrião de Mamíferos/patologia , Desenvolvimento Embrionário , Fertilização In Vitro/métodos , Zigoto/patologia , Animais , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Embrião de Mamíferos/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Endogâmicos ICR , Zigoto/metabolismo
14.
Theriogenology ; 174: 36-46, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34416562

RESUMO

The removal of the zona pellucida has been used to improve the in vitro development of domestic cat embryos generated by IVF and SCNT. However, the in vivo development of domestic cat embryos generated without the zona pellucida has not been evaluated. The objective of this study was to evaluate the effects of zona pellucida removal on the in vitro and in vivo development of domestic cat embryos generated by IVF. For this purpose, two experimental groups were created: 1) domestic cat embryos cultured in vitro (Zona-intact group, ZI) and 2) domestic cat embryos cultured in vitro without the zona pellucida (Zona-free group, ZF). Domestic cat embryos were generated by IVF and cultured in vitro for 8 days. In the ZF group, the zona pellucida was removed after IVF, and embryos were cultured using the well of the well system (WOW). Cleavage, morula and blastocyst rates were evaluated in both groups. The diameter and total cell number of blastocysts were assessed. Relative expression of pluripotency (OCT4, SOX2 and NANOG), differentiation (CDX2 and GATA6) and apoptotic markers (BAX and BCL2) was evaluated in blastocysts. Finally, to evaluate in vivo development, embryos at days 5, 6 and 7 of development were transferred into recipient domestic cats, and ultrasonography was performed to evaluate implantation. No differences were observed in the cleavage, morula or blastocyst rates between embryos from the ZI and ZF groups. The diameter (mean ± SD) of blastocysts from the ZF group was greater (253.4 ± 83.3 µm) than that from the ZI group (210.5 ± 78.5 µm). No differences were observed in the relative expression of OCT4, CDX2 or GATA6. However, the relative expression of SOX2 and NANOG was significantly reduced in ZF blastocysts compared to ZI blastocysts. Furthermore, the relative expression of BAX was higher in ZF blastocysts than in ZI blastocysts. Finally, four pregnancies were confirmed after the transfer of ZI embryos (n = 110). However, no pregnancies were observed after the transfer of ZF embryos at the morula or blastocyst stage (n = 56). In conclusion, domestic cat embryos cultured without the zona pellucida were able to develop in vitro until the blastocyst stage. However, the removal of the zona pellucida negatively affected the gene expression of pluripotency and apoptosis markers, and ZF embryos were unable to implant. This might indicate that the removal of the zona pellucida is detrimental for the implantation and in vivo development of domestic cat embryos.


Assuntos
Blastocisto , Zona Pelúcida , Animais , Gatos , Implantação do Embrião , Feminino , Fertilização In Vitro/veterinária , Expressão Gênica , Mórula , Gravidez
15.
Science ; 373(6552)2021 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-34437124

RESUMO

Oocytes mature in a specialized fluid-filled sac, the ovarian follicle, which provides signals needed for meiosis and germ cell growth. Methods have been developed to generate functional oocytes from pluripotent stem cell-derived primordial germ cell-like cells (PGCLCs) when placed in culture with embryonic ovarian somatic cells. In this study, we developed culture conditions to recreate the stepwise differentiation process from pluripotent cells to fetal ovarian somatic cell-like cells (FOSLCs). When FOSLCs were aggregated with PGCLCs derived from mouse embryonic stem cells, the PGCLCs entered meiosis to generate functional oocytes capable of fertilization and development to live offspring. Generating functional mouse oocytes in a reconstituted ovarian environment provides a method for in vitro oocyte production and follicle generation for a better understanding of mammalian reproduction.


Assuntos
Células-Tronco Embrionárias Murinas/fisiologia , Oócitos/fisiologia , Oogênese , Folículo Ovariano/citologia , Animais , Técnicas de Cultura de Células , Diferenciação Celular , Desenvolvimento Embrionário , Feminino , Fertilização In Vitro , Masculino , Mesoderma/citologia , Mesoderma/fisiologia , Camundongos , Camundongos Endogâmicos ICR , Células-Tronco Embrionárias Murinas/citologia , Oócitos/citologia , Folículo Ovariano/embriologia , Folículo Ovariano/fisiologia , RNA-Seq , Fator Esteroidogênico 1/genética , Fator Esteroidogênico 1/metabolismo , Transcriptoma
16.
FASEB J ; 35(9): e21812, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34411354

RESUMO

Blastocysts hatch from the zona pellucida (ZP) to enable implantation into the uterine endometrial epithelium, but little is known regarding the effect of hatching sites on pregnancy outcomes. Murine hatching embryos were categorized into five groups based on initial trophectoderm projection (TEP)/ZP position corresponding to the inner cell mass center. In blastocysts (3.5 dpc) post-12 hours in vitro culture, TEP rates of A-site (44.4%) and B-site (38.6%) embryos were higher than those of C-site (12.5%) and D-site (3.1%) embryos, while the O-site (1.4%) was the lowest (P < .05). Post-ET A-site (55.6%) and B-site (65.6%) birth rates were higher than those of C-site embryos (21.3%) and controls (P < .05). Furthermore, live birth rate of B-site embryos remained higher than C-site embryos (68.8% vs 31.3%; P < .05) when both were transferred into the same recipients. Different TEP site blastocysts exhibited different implantation competences: the implantation rate of C-site embryos was lower than that of both A- and B-site groups (67.7% vs 84.3% and 83.2%, respectively; P < .05) at 2 days post-ET. C-site embryos also had a distinctly higher ratio of developmental defects (47.5%) than A- and B-site embryos (22.5% and 14.6%, respectively), with implantation failure mainly associated with poor birth rate, a finding corroborated by differential gene expression analysis such as LIF, LIFR, and S100a9. Surprisingly, acidified Tyrode's solution (AAH)-treated B-site blastocysts had a significantly increased birth rate (77.1%) than C-site (55.3%) and controls (43.4%). Site specificity and differential gene expression during embryo hatching can be applied in ART screening. More importantly, assisted hatching by AAH is effective and feasible for improving pregnancy and term development, particularly at the B-site, for humans and in animal husbandry.


Assuntos
Coeficiente de Natalidade , Blastocisto/citologia , Implantação do Embrião , Trofoblastos/citologia , Zona Pelúcida/metabolismo , Animais , Transferência Embrionária , Feminino , Fertilização In Vitro , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Camundongos , Gravidez , Resultado da Gravidez , Útero/citologia
17.
Zool Res ; 42(5): 562-573, 2021 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-34355875

RESUMO

Inositol requiring mutant 80 (INO80) is a chromatin remodeler that regulates pluripotency maintenance of embryonic stem cells and reprogramming of somatic cells into pluripotent stem cells. However, the roles and mechanisms of INO80 in porcine pre-implantation embryonic development remain largely unknown. Here, we show that INO80 modulates trophectoderm epithelium permeability to promote porcine blastocyst development. The INO80 protein is highly expressed in the nuclei during morula-to-blastocyst transition. Functional studies revealed that RNA interference (RNAi)-mediated knockdown of INO80 severely blocks blastocyst formation and disrupts lineage allocation between the inner cell mass and trophectoderm. Mechanistically, single-embryo RNA sequencing revealed that INO80 regulates multiple genes, which are important for lineage specification, tight junction assembly, and fluid accumulation. Consistent with the altered expression of key genes required for tight junction assembly, a permeability assay showed that paracellular sealing is defective in the trophectoderm epithelium of INO80 knockdown blastocysts. Importantly, aggregation of 8-cell embryos from the control and INO80 knockdown groups restores blastocyst development and lineage allocation via direct complementation of the defective trophectoderm epithelium. Taken together, these results demonstrate that INO80 promotes blastocyst development by regulating the expression of key genes required for lineage specification, tight junction assembly, and fluid accumulation.


Assuntos
ATPases Associadas a Diversas Atividades Celulares/metabolismo , Blastocisto/fisiologia , Cromatina/metabolismo , Proteínas de Ligação a DNA/metabolismo , Mórula/fisiologia , Suínos , ATPases Associadas a Diversas Atividades Celulares/genética , Animais , Proteínas de Ligação a DNA/genética , Técnicas de Cultura Embrionária/veterinária , Fertilização In Vitro , Regulação da Expressão Gênica/fisiologia , Oócitos/fisiologia , Permeabilidade
18.
Niger J Clin Pract ; 24(8): 1159-1163, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34397024

RESUMO

Objective: Our aim was to compare the efficacy of two triggering method one with dual triggering with gonadotropin-realising hormon (GnRH) agonist plus standard dosage human chorionic gonadotropin (hCG) and the other with hCG only for final oocyte maturation on oocyte/follicle ratio and pregnancy rates in normoresponders in GnRH antagonist cycles in invitro fertilization-intrastoplasmic sperm injection (IVF-ICSI). Material Methods: In this retrospective study, all patients underwent GnRH antagonist protocol. When at least ≥3 follicles reached ≥17 mm diameter, 116 patients received dual trigger with GnRH agonist plus hCG (1mg Leuprolide acetate plus 10.000 IU uhCG) and 178 patients received uhCG (10.000 IU u hCG) for final oocyte maturation. All follicles ≥10 mm diameter were aspirated. Number of oocytes and metaphase II oocytes retrieved per aspirated follicles, implantation rate, and clinical pregnancy rate per cycle was recorded. Results: There was no statistically significant difference in terms of metaphase II oocyte ratio per aspirated follicle, implantation rate and clinical pregnancy rate between the dual trigger group and hCG only group (45.7% vs. 51%; 35.4% vs.30.3% and 45%vs. 40% respectively). Oocyte/ follicle ratio was significantly higher in dual trigger group (68.2%vs 63.8% p=0,028). Conclusions: Dual triggering in normal responders with a GnRH-agonist and a standard dosage of hCG is superior to hCG only protocol in terms of oocyte/follicle ratio but does not improve metaphase II oocyte, implantation and clinical pregnancy rates in GnRH-antagonist cycles. Dual triggering method may be beneficial in patients with immature oocytes and emty follicle syndrome.


Assuntos
Fertilização In Vitro , Indução da Ovulação , Gonadotropina Coriônica , Feminino , Hormônio Liberador de Gonadotropina , Humanos , Oócitos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Injeções de Esperma Intracitoplásmicas
19.
Int J Mol Sci ; 22(15)2021 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-34361119

RESUMO

Developmental arrest of the preimplantation embryo is a multifactorial condition, characterized by lack of cellular division for at least 24 hours, hindering the in vitro fertilization cycle outcome. This systematic review aims to present the molecular drivers of developmental arrest, focusing on embryonic and parental factors. A systematic search in PubMed/Medline, Embase and Cochrane-Central-Database was performed in January 2021. A total of 76 studies were included. The identified embryonic factors associated with arrest included gene variations, mitochondrial DNA copy number, methylation patterns, chromosomal abnormalities, metabolic profile and morphological features. Parental factors included, gene variation, protein expression levels and infertility etiology. A valuable conclusion emerging through critical analysis indicated that genetic origins of developmental arrest analyzed from the perspective of parental infertility etiology and the embryo itself, share common ground. This is a unique and long-overdue contribution to literature that for the first time presents an all-inclusive methodological report on the molecular drivers leading to preimplantation embryos' arrested development. The variety and heterogeneity of developmental arrest drivers, along with their inevitable intertwining relationships does not allow for prioritization on the factors playing a more definitive role in arrested development. This systematic review provides the basis for further research in the field.


Assuntos
Blastocisto/patologia , Embrião de Mamíferos/patologia , Desenvolvimento Embrionário , Fertilização In Vitro , Humanos
20.
Zhonghua Yi Xue Za Zhi ; 101(28): 2228-2232, 2021 Jul 27.
Artigo em Chinês | MEDLINE | ID: mdl-34333936

RESUMO

Objective: To evaluate the clinical effect of oral contraceptive (OC) pretreatment on the outcome of gonadotropin releasing hormone antagonist (GnRH-a) protocol in patients with non-polycystic ovary syndrome. Methods: From January 2017 to May 2019, a total of 436 patients undergoing in vitro fertilization and embryo transfer/Intracytoplasmic sperm injection (IVF-ET/ICSI) treatment in Peking University First Hospital reproductive center clinic were included in this retrospective cohort study. A total of 144 patients (147 cycles) used OC pretreatment prior to GnRH-a protocol and 292 patients (306 cycles) used GnRH-a protocol without OC pretreatment. The drug usage as well as pregnant outcomes between groups were examined. The primary outcome was the cumulative clinical pregnancy rate of oocyte retrieval cycle and the secondary outcome included the number of oocytes, MⅡ oocytes, embryos and clinical pregnancy rate of fresh embryo transfer cycle. Results: The median ages (and Q1, Q3) of OC pretreatment group and non-OC group were 33 (30,36) and 34 (30,38) years old, respectively. The number of MⅡ oocytes was higher in OC pretreatment group (7/9) than in non-OC group (6/8) (P=0.002). The significant difference were not found in the cumulative clinical pregnancy rate of each oocyte retrieval cycle (61.7% vs 54.6%), the clinical pregnancy rate of fresh embryo transfer cycle (34.4% vs 35.6%), and the number of oocytes (9 vs 8) and embryos (6 vs 6) between groups. Conclusion: Our findings suggest that compared to non-OC pretreatment group, pretreatment with OC is associated with more MⅡ oocytes, and with an increasing trend of the cumulative clinical pregnancy rate in non-polycystic ovary syndrome patients undergoing fresh IVF-ET/ICSI.


Assuntos
Anticoncepcionais Orais , Indução da Ovulação , Feminino , Fertilização In Vitro , Hormônio Liberador de Gonadotropina , Antagonistas de Hormônios , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
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