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1.
Zhonghua Yi Xue Za Zhi ; 99(46): 3627-3632, 2019 Dec 10.
Artigo em Chinês | MEDLINE | ID: mdl-31826584

RESUMO

Objective: To find the best strategy of embryo transfer, so as to provide theoretical basis for improving the clinical outcomes of in vitro fertilization-Embryo transfer (IVF-ET), we investigate the blastocyst culture of surplus cleavage-stage embryos after D3 embryo transfer and the prediction of clinical outcomes with or without blastocyst formation. Methods: A retrospective study was conducted on 3 568 patients who underwent IVF-ET in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2016 to May 2018, whotransplanted two embryos in D3 with blastocyst culture of surplus cleavage-stage embryos, according to their age, they were divided into three groups: <35 years old group, 35-38 years old group, and>38 years old group.And according to the presence or absence of blastocyst formation, they were also divided into two subgroups: blastocyst formation group and non-blastocyst formation group. The embryo development and clinical outcomes in each group were compared. Results: (1) Comparisons of the embryo development in the three age groups with the first cycle. The total fertilization rate, cleavage rate and high quality embryo rate of the blastocyst formation group in the three groups were higher than those in the non-blastocyst formation group, P<0.05; In<35 years old group, the embryo utilization rate (75.0% vs 70.6%), pregnancy rate (74.9% vs 70.3%), planting rate (53.6% vs 48.6%), delivery rate (66.7% vs 61.1%) and live birth rate (66.5% vs 61.0%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. (2) Comparisons of embryo development in the three age groups with multiple cycles (≥2 cycles). In<35 years old group, the total fertilization rate (75.0% vs 70.6%),delivery rate (62.7% vs 43.8%) and live birth rate (62.7% vs 43.8%) of the blastocyst formation group were significantly higher than those in the non-blastocyst formation group, P<0.05; In>38 years old group, the pregnancy rate (56.3% vs 25.8%), implantation rate (34.4% vs 14.5%), delivery rate (43.8% vs 11.3%), live birth rate (43.8% vs 11.3%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. Conclusions: The results of blastocyst culture in different groups can predict the outcomes of embryo transfer in D3. For patients<35 years old with the first cycle, the clinical outcomes of the blastocyst formation group after D3 embryo transfer is better than that of the non-blastocyst formation group. For Patients with multiple cycles (≥2 cycles),the clinical outcomes of the embryo formation group is superior to that of the non-blastocyst formation group<35 years old or>38 years old.


Assuntos
Blastocisto , Fase de Clivagem do Zigoto , Transferência Embrionária , Gravidez Múltipla , Adulto , Feminino , Fertilização In Vitro , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
2.
Medicine (Baltimore) ; 98(38): e17329, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31568019

RESUMO

Thus far, all clinical trials evaluating the efficacy of embryo transfer strategies have selectively delayed the first frozen embryo transfer (FET) by at least 1 menstrual cycle. Nevertheless, this approach, which is based solely on clinical experience, may create unnecessary psychological stress on infertile patients who are anxious to conceive as soon as possible. This study aimed to investigate whether the time interval between oocyte retrieval and subsequent FET affects reproductive outcomes.We implemented a large retrospective cohort study in a single assisted reproductive technology (ART) unit at a university-based hospital, including 1540 autologous FET cycles performed in freeze-all cycles. The beginning of the FET was classified as either 'cycle 1' (performing FET within the first menstrual cycle) or 'cycle ≥2' (performing FET after one or more menstrual cycles). Live birth rate (LBR) was the primary outcome of our study.The mean interval for 'cycle 1' and 'cycle ≥2' FETs was 25.72 ±â€Š5.10 days and 75.33 ±â€Š24.85 days, respectively (P < .001). The type of controlled ovarian hyperstimulation (COH) and endometrial preparation protocols differed significantly between groups (P = .008 and P = .004, respectively). However, FET groups were similar in many ways. Univariate analysis showed that there was no significant difference in LBR between the different cycles (33.1% after 'cycle 1' FET vs 34.2% after 'cycle ≥2' FET, P = .68). To evaluate whether LBR remained unchanged after adjustment for potential confounders, we performed multivariate logistic regression. FET timing had no significant impact on LBR in the first FET (odds ratio [OR]: 1.06, 95% confidence interval [CI]: 0.80-1.39).In accordance with the present study, it might not be necessary for clinicians to wait more than 1 menstrual cycle before performing FET. This allows us to reduce otiose deferment in FET, without adversely affecting reproductive outcomes.


Assuntos
Fase de Clivagem do Zigoto , Criopreservação , Transferência Embrionária/métodos , Ciclo Menstrual , Adulto , Feminino , Humanos , Gravidez , Resultado da Gravidez , Técnicas de Reprodução Assistida , Estudos Retrospectivos
4.
Medicina (Kaunas) ; 55(6)2019 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-31226849

RESUMO

BACKGROUND AND OBJECTIVE: During the last few years, a trend has been noted towards embryos being transferred at the blastocyst stage, which has been associated with improved rates regarding implantation and clinical pregnancy in comparison to cleavage stage embryo transfers. There is a limited number of studies investigating this notion in oocyte donation cycles employing cryopreserved embryos. The aim of this study is to evaluate the implantation potential and clinical pregnancy rates between the day 3 cleavage stage and blastocyst stage embryo transfers in oocyte donation cycles employing vitrified embryos. METHODS: This is a retrospective evaluation of oocyte donation frozen-thawed transfers completed in our clinic from January 2017 to December 2017. Intracytoplasmic sperm injection was conducted for all oocytes. Following fertilization, all embryos were cryopreserved either at the cleavage or blastocyst stage. Embryo transfer of two embryos was performed under direct sonographic guidance in all cases. Results: Our results confirmed a 55.6% clinical pregnancy (CP) resulting from day 3 embryo transfers, a 68.8% CP from day 5, and 71.4% CP from day 6. Significantly improved pregnancy rates were related to embryo transfers at the blastocyst stage when compared to cleavage stage transfers (68.9% and 55.6% respectively, p = 0.016). The risk with regards to multiple pregnancies was similar. CONCLUSION: Our findings indicate that in oocyte donation cycles employing vitrified embryos, embryo transfer at the blastocyst stage is accompanied with a significant improvement in pregnancy rates and merits further investigation.


Assuntos
Transferência Embrionária/métodos , Fatores de Tempo , Resultado do Tratamento , Adulto , Fase de Clivagem do Zigoto , Transferência Embrionária/tendências , Feminino , Humanos , Doação de Oócitos/métodos , Gravidez , Estudos Retrospectivos
5.
Gynecol Endocrinol ; 35(9): 792-795, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30971141

RESUMO

To compare the pregnancy and obstetric outcomes following single cleavage-stage embryo transfer (SCT) and single blastocyst transfer (SBT) using time-lapse imaging (TLI), a total of 2066 normally fertilized and cleaved embryos from 233 patients were divided into Day 3 SCT group (n = 171) and Day 5 SBT group (n = 62) according to patient's willingness. Embryo selection criteria were based on embryo cleavage patterns, timing parameters, and blastocyst quality. The pregnancy and obstetric outcomes of each group were evaluated. There were no statistically significant differences with regard to pregnancy outcomes including the implantation rate, early abortion rate, ongoing pregnancy rate and live birth rate, and obstetric outcomes including preterm birth rate, gestational week, birth height, birth weight and fetal malformation rate between SCT group and SBT group. SBT group had significantly higher monozygotic twinning (MZT) rates than SCT group (6.98% vs. 0, p < .05). Although not statistically significant, there was a trend of higher proportion of male-to-female sex ratio at birth in SBT group than SCT group (1.38 vs. 1.05). Based on the combination of cleavage patterns and timing parameters, SCT may be an alternative to SBT because it can provide similar pregnancy and obstetric outcomes and meanwhile lower monozygotic twinning rates.


Assuntos
Fase de Clivagem do Zigoto/fisiologia , Transferência Embrionária , Resultado da Gravidez/epidemiologia , Transferência de Embrião Único , Imagem com Lapso de Tempo , Adulto , Fase de Clivagem do Zigoto/citologia , Parto Obstétrico/métodos , Parto Obstétrico/estatística & dados numéricos , Técnicas de Diagnóstico Obstétrico e Ginecológico , Transferência Embrionária/métodos , Transferência Embrionária/estatística & dados numéricos , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Estudos Retrospectivos , Transferência de Embrião Único/métodos , Transferência de Embrião Único/estatística & dados numéricos , Imagem com Lapso de Tempo/métodos , Imagem com Lapso de Tempo/estatística & dados numéricos
6.
Curr Med Sci ; 39(1): 118-121, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30868500

RESUMO

To investigate the developmental potential and clinical value of embryos with abnormal cleavage rate, a retrospective analysis was performed on 66 635 2-prokaryotic (2PN) and 1-pronuclear (1PN) embryos. The embryos were given conventionally in vitro fertilization (IVF) treatment and continuously cultured on the day 3 (D3) at the Reproductive Medicine Center, Tongji Medical College, Huazhong University of Science and Technology from January 2016 to December 2017. The embryos were separated into the day-2 (D2) undivided group with 106 cases, the arrested development group with 3482 cases, the blastomere reduction group with 541 cases, and the control group with 62 506 cases, respectively. The blastocyst utilization rates of these three abnormal groups were 2.83%, 10.86% and 6.84%, respectively, which were significantly different from that in control group (39.46%). Furthermore, 2 cases of anabiosis and 1 case of live birth were found in D2 undivided group. In arrested development group, there were 55 cases of anabiosis, 11 cases of clinical pregnancy in single-embryo transplantation (including 6 cases of live birth), and 25 cases of clinical pregnancy in combination with one normal embryo transplantation (including 23 cases of live births and 15 cases of dizygotic twins under B-ultrasound). There were 13 case of anabiosis in blastomere reduction group: there was 1 case of single embryo transplantation and clinical pregnancy was obtained; there were also 6 cases of clinical pregnancy in combination with one single normal embryo transplantation (including 5 cases of live births and 2 cases of dizygotic twins under B-ultrasound). In conclusion, embryos with abnormal cleavage rate still have the potential to continue to develop, and have certain blastocyst utilization rate and live birth.


Assuntos
Blastômeros/citologia , Fase de Clivagem do Zigoto/patologia , Desenvolvimento Embrionário , Nascimento Vivo/epidemiologia , Técnicas de Cultura Embrionária , Implantação do Embrião , Feminino , Fertilização In Vitro , Humanos , Gravidez , Taxa de Gravidez , Estudos Retrospectivos
7.
Cells ; 8(2)2019 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-30781491

RESUMO

Reproductive ageing in women, particularly after the age of 35, is associated with an exponential increase in the proportion of chromosomally abnormal oocytes produced. Several hypotheses have attempted to explain this observation, including the 'limited oocyte pool' hypothesis and the 'two-hit' hypothesis, the latter explaining that a depletion in oocyte quality with age results from the multiple opportune stages for errors to occur in meiosis. Recently however, the telomere theory of reproductive ageing in women has been proposed. This suggests that shortened telomeres in oocytes of women of advanced maternal age render oocytes unable to support fertilization and embryogenesis. Despite a credible rationale for the telomere theory of reproductive ageing in women, very few studies have assessed telomere length directly in human oocytes or preimplantation embryos. Therefore, we directly assessed relative telomere length in first polar bodies and blastomeres from cleavage stage (day 3) embryos. In both cell types we tested the hypothesis that (1) older women have shorter telomeres and (2) chromosomally abnormal (aneuploid) gametes/embryos have shorter telomeres. In all cases, we found no evidence of altered telomere length associated with age-related aneuploidy.


Assuntos
Envelhecimento/fisiologia , Aneuploidia , Fase de Clivagem do Zigoto/metabolismo , Embrião de Mamíferos/metabolismo , Corpos Polares/metabolismo , Reprodução , Análise de Célula Única , Telômero/metabolismo , Adulto , Blastômeros/metabolismo , Humanos , Idade Materna , Homeostase do Telômero
8.
Gynecol Endocrinol ; 35(8): 665-668, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30806103

RESUMO

Today, most IVF programs have moved to blastocyst transfer but there is still uncertainty regarding when to transfer if there are only one or two embryos at the cleavage stage. The aim of this study was to compare the pregnancy rate of day 3 transfers vs. blastocyst stage transfers in patients who had only one or two embryos on day 3. We conducted a retrospective study of 102 patients with one or two cleavage stage embryos that had their embryos transferred on day 3 and 429 patients had their embryos cultured to day 5 for transfer. The number of mature oocytes (4.0 vs 4.6, p = NS) and number of cleavage stage embryos on day 3 was similar in the two groups (1.3 vs. 1.5, p = NS). The clinical pregnancy rate per retrieval (22% vs. 24.6%, p= NS) and the ongoing pregnancy rate per retrieval (20% vs. 20.2%, p = NS) was comparable between the groups. Fifty seven (13.2%) of the patients had cleavage embryo arrest and did not have an embryo to transfer on day 5. We conclude that the cumulative pregnancy rate is the same for patients with 1-2 cleavage stage embryos regardless of whether the embryo is transferred on day 3 or day 5.


Assuntos
Fase de Clivagem do Zigoto/fisiologia , Transferência Embrionária/métodos , Adulto , Blastômeros/citologia , Fase de Clivagem do Zigoto/citologia , Estudos de Coortes , Transferência Embrionária/normas , Embrião de Mamíferos , Feminino , Humanos , Gravidez , Taxa de Gravidez , Gravidez de Gêmeos , Estudos Retrospectivos , Transferência de Embrião Único/métodos , Fatores de Tempo , Gêmeos
9.
J Reprod Dev ; 65(3): 245-250, 2019 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-30773507

RESUMO

Piezo-actuated intracytoplasmic sperm injection (Piezo-ICSI) is used as an efficient in vitro fertilization method with various animals. With this method, elongated spermatids are collected from testicular tissues and are easier to obtain from animals that unexpectedly die than ejaculate sperm. Additionally, elongated spermatid injection often results in the development of embryos and offspring. To develop assisted reproductive techniques (ARTs) for domestic cats, we examined the effects of oocyte activation on cleavage and embryo development after Piezo-ICSI with motile sperm (experiment 1) and after Piezo-ICSI with either testicular sperm or elongated spermatids (experiment 2). In experiment 1, the proportions of cleaved embryos, morulas, and blastocysts following Piezo-ICSI with ethanol activation were significantly higher (P < 0.05) than in the non-activated groups. However, the proportion of blastocysts and the blastocyst quality did not differ significantly (P > 0.05) between the ethanol-activated and non-activated groups. In experiment 2, the cleavage frequencies of oocytes after Piezo-ICSI of testicular sperm or elongated spermatids and ethanol activation were higher (P < 0.05) than that of oocytes in the non-activated group, but the occurrence of blastocyst formation and quality of blastocysts did not differ between the activated and non-activated groups. In summary, cat embryos can be produced by Piezo-actuated microinjection of elongated spermatids. Ethanol activation increased the frequency of cleavage, but it affected neither the occurrence of blastocyst development nor the quality of blastocysts. These results represent an expansion in the repertoire of ARTs that are potentially applicable to both domestic and endangered species of cats.


Assuntos
Oócitos/citologia , Injeções de Esperma Intracitoplásmicas/veterinária , Espermátides/fisiologia , Espermatozoides/fisiologia , Animais , Blastocisto/citologia , Gatos , Fase de Clivagem do Zigoto , Criopreservação , Desenvolvimento Embrionário , Feminino , Fertilização In Vitro , Masculino , Microinjeções , Ovário/citologia , Testículo/citologia
10.
Methods Mol Biol ; 1920: 393-406, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30737705

RESUMO

The division patterns of early invertebrate and vertebrate embryos are key to the specification of cell fates and embryo body axes. We here describe a generic computational modeling method to quantitatively test mechanisms which specify successive division position and orientation of eggs and early blastomeres in 3D. This approach should serve to motivate and guide future experimental work on the mechanisms controlling early embryo morphogenesis.


Assuntos
Fase de Clivagem do Zigoto , Desenvolvimento Embrionário/fisiologia , Modelos Biológicos , Diferenciação Celular , Divisão Celular , Técnicas de Cultura Embrionária , Imageamento Tridimensional , Microscopia Confocal
11.
J Reprod Dev ; 65(2): 163-170, 2019 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-30700637

RESUMO

Lycium barbarum polysaccharide (LBP) exhibits multiple pharmacological and biological effects, including displaying antioxidant and cytoprotective properties. The current study investigated the effects of LBP-supplemented culture medium on mitochondrial distribution, mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) production, mitochondrial deoxyribonucleic acid (mtDNA) copy number, reactive oxygen species (ROS) accumulation, and development of previously-cryopreserved murine two-cell embryos. Results indicate that LBP enhances development of such embryos, and that potential mechanisms include: (1) mitochondrial function enhancement via altering mitochondrial distribution and increasing MMP, ATP production, mtDNA copy number, and expression of genes involved in mitochondrial biogenesis and energy metabolism (NAD-dependent deacetyltransferase sirtuin-1 (SIRT1) and phosphorylated adenosine monophosphate-activated protein kinase (pAMPK)); (2) down-regulation of ROS generation and enhanced expression of the antioxidant genes glutathione peroxidase 4 (GPX4) and superoxide dismutase 1 (SOD1), thereby increasing embryo oxidative stress tolerance; and (3) increased expression of B-cell lymphoma-2 (BCL2), a critical gene for cell survival and embryo development. These results demonstrate that LBP improves development of previously-cryopreserved murine two-cell embryos via restoration of mitochondrial function and down-regulated generation of ROS.


Assuntos
Criopreservação , Medicamentos de Ervas Chinesas/farmacologia , Embrião de Mamíferos , Desenvolvimento Embrionário/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fase de Clivagem do Zigoto , Criopreservação/veterinária , Regulação para Baixo/efeitos dos fármacos , Técnicas de Cultura Embrionária , Feminino , Masculino , Camundongos , Mitocôndrias/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Gravidez
12.
Reprod Domest Anim ; 54(5): 804-807, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30786073

RESUMO

This study examines the presence of activin IIA and IIB receptors (ActR-IIA and ActR-IIB) by Western blotting and immunocytochemistry in immature and IVM-oocytes, 2 to 8-cells embryos and blastocysts from prepubertal goats. Western blotting revealed that activin receptors are synthesized during oocyte maturation and embryo development. In the immunocytochemistry experiments, no immunostaining for either receptor was detected in oocytes while both receptors were immunolabelled in all the cells of cleaved embryos. In blastocysts, while ActR-IIA expression appeared evenly distributed in the two cell lineages, inner cell mass and trophectoderm, the ActR-IIB immunosignal was restricted mainly to the inner cell mass. Our findings reveal the presence of activin type II receptors (ActR-IIA and ActR-IIB) in in vitro matured prepubertal goat oocytes and blastocyst-stage embryos. The expression of these receptors could be a key factor in understanding differences between competent and incompetent oocytes.


Assuntos
Receptores de Activinas Tipo II/genética , Desenvolvimento Embrionário , Fertilização In Vitro/veterinária , Oócitos/metabolismo , Animais , Blastocisto/fisiologia , Fase de Clivagem do Zigoto , Feminino , Expressão Gênica , Cabras
13.
Lasers Med Sci ; 34(6): 1137-1145, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30627926

RESUMO

Assisted hatching (AH) is initially developed to provide an artificial manipulation of the zona pellucida (ZP) to help embryos hatch and improve the capacity of the embryos to implant. However, these effects remain unclear and controversial because of variation in patient characteristics, and it is critical to ascertain the indications for AH and to identify those patients who might benefit from AH. Here, this study aimed to assess the effect of laser-assisted zona thinning hatching technology (LAH) during the frozen-thawed D3 embryos on pregnancy outcomes in patients with previous repeated failures in vitro fertilization-embryo transfer (IVF-ET). To the best of our knowledge, these relationships have not been previously investigated. A retrospective cohort analysis was carried out. Infertility patients with previous repeated failure who underwent assisted reproductive therapy at our in vitro fertilization (IVF) center from May 2014 to May 2016 were enrolled. A total of 415 cleavage FET cycles (225 in the LAH group and 190 in the control group) were analyzed. Clinical outcomes including clinical pregnancy, implantation, live birth, miscarriage, and multiple gestation rates after transfer were compared between the LAH and control groups. The clinical pregnancy (49.3% versus 38.9%) and implantation rates (31.2% versus 24.6%) were significantly higher for the LAH group than the control group (P < 0.05). The live birth (44.8% versus 35.8%), multiple pregnancy (32.4% versus 31.0%), preterm birth (22.8% versus 17.1%), miscarriage (7.2% versus 5.4%), and ectopic rates (1.9% versus 0%) did not differ significantly between the two groups (P > 0.05). This study showed that LAH via zona pellucida (ZP) thinning significantly improves clinical outcomes, particularly clinical pregnancy and implantation rates, associated with FET cycles among patients with previous repeated failure.


Assuntos
Fase de Clivagem do Zigoto/transplante , Criopreservação , Transferência Embrionária , Lasers , Resultado da Gravidez , Técnicas de Reprodução Assistida , Adulto , Implantação do Embrião , Feminino , Humanos , Recém-Nascido , Gravidez , Estudos Retrospectivos , Resultado do Tratamento
14.
Reprod Domest Anim ; 54(2): 300-308, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30325531

RESUMO

The purpose of the present study was to assess the effect of crocin supplementation during oocyte maturation on the antioxidant defence and anti-apoptotic ability and subsequent developmental competence of porcine oocytes. Oocytes were cultured in media containing 0, 300, 400 or 500 µg/ml of crocin. Upon maturation, the maturation rates, reactive oxygen species (ROS) and glutathione (GSH) levels, mRNA expression of genes (SOD, CAT, GPx, Bcl-2, BAX and Caspase3), expression of cleaved caspase3 and subsequent embryo cleavage rates were measured. Results indicated that the maturation rate of the 400 µg/ml group was 86.80% (p < 0.01). The ROS concentration of the 500 µg/ml group was the lowest (p < 0.01). The GSH concentration of the 400 µg/ml group was the highest (p < 0.01). The SOD, CAT and GPx mRNA expression levels were the highest in the 300, 400 and 500 µg/ml groups, respectively, with the expression levels of all genes being significantly higher than that of the control group (p < 0.01). The Bcl-2/BAX mRNA expression ratio in 400 and 500 µg/ml groups significantly higher than other groups and significantly decreased caspase3 expression level (p < 0.01). The expression level of cleaved caspase3 in the 500 µg/ml treatment group was the lowest, significantly lower than that of the control group (p < 0.01). The cleavage rate of the 400 µg/ml group was 62.50% (p < 0.01). These experimental results show that the supplementation of in vitro culture medium with 400 µg/ml of crocin significantly enhanced the antioxidant defence and anti-apoptotic ability and subsequent cleavage rate of porcine embryo.


Assuntos
Blastocisto/efeitos dos fármacos , Carotenoides/farmacologia , Fase de Clivagem do Zigoto/efeitos dos fármacos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Caspase 3/metabolismo , Técnicas de Cultura Embrionária/veterinária , Desenvolvimento Embrionário , Feminino , Expressão Gênica , Glutationa/metabolismo , Técnicas de Maturação in Vitro de Oócitos/métodos , Partenogênese , Espécies Reativas de Oxigênio/metabolismo , Suínos
15.
Int J Biol Macromol ; 123: 931-938, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30452988

RESUMO

Tetraspanins are multifunctional molecules located in specific microdomains on the plasma membrane. Thanks to their ability to form networks with other proteins they can participate in many cellular functions. Tetraspanins are part of the interactive network in gametes; however, their precise role in fertilization is not yet clear. The aim of this study was to compare the localization of CD9 and CD81 tetraspanins during oocyte maturation and early development of the embryos in bovine and porcine model. CD9 was detected on the oocyte plasma membrane and vesicles in the perivitelline space of bovine oocytes and embryos. We suggest that CD9 could be a component involved in transzonal projections. Based on the results of in vitro fertilization assay, CD9 and CD81 seem to be part of a more complex fusion network on the plasma membrane of bovine oocytes. On the other hand, both tetraspanins showed a clustered expression pattern on the plasma membrane and inner margin of zona pellucida (ZP) in porcine oocytes and embryos. We found a new species-specific pattern of CD9 and CD81 distribution in ZP which could reflect their specialized role in processes associated with cell adhesion and intercellular communication upon fertilization.


Assuntos
Embrião de Mamíferos/metabolismo , Oócitos/metabolismo , Tetraspanina 28/metabolismo , Tetraspanina 29/metabolismo , Animais , Anticorpos/farmacologia , Bovinos , Linhagem Celular , Fase de Clivagem do Zigoto/citologia , Fase de Clivagem do Zigoto/efeitos dos fármacos , Embrião de Mamíferos/citologia , Feminino , Fertilização In Vitro/efeitos dos fármacos , Metáfase/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Oócitos/citologia , Partenogênese/efeitos dos fármacos , Suínos
16.
Eur J Obstet Gynecol Reprod Biol ; 233: 107-113, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30590226

RESUMO

OBJECTIVE: To determine the effect of embryonic factors on serum beta human chorionic gonadotropin (ß-hCG) levels in pregnancy and live birth resulting after a single fresh cleavage embryo and blastocyst transfer. STUDY DESIGN: This was a retrospective cohort study conducted at a tertiary care hospital. All fresh single embryo transfers (sETs) between September 2011 and December 2016 were included. The correlation analysis was performed to determine the association of embryo morphological parameters on mean serum ß-hCG levels on day 12 after the transfer of a fresh single cleavage embryo and a fresh single blastocyst embryo. RESULTS: Out of a total of 455 fresh sETs, 60 positive ß-hCG results after the transfer of a single fresh cleavage-stage embryo and 82 after the transfer of a single fresh blastocyst. The mean ß-hCG level resulting from a single fresh blastocyst ET was 371.7 ± 52.7 IU/L, which was similar to the mean ß-hCG level resulting from a cleavage ET (314.5 ± 36.9 IU/L) (p = .70). Interestingly, serum ß-hCG levels resulting from a single fresh blastocyst ET showed a correlation with day 5 blastocoele expansion, trophectoderm cell number and blastocyst quality score in ongoing pregnancy (r = .33, p = .02; r = .29, p = .04; and r = .31, p = .03, respectively). Moreover, day 5 blastocoele expansion and blastocyst quality score showed a correlation with the serum ß-hCG levels resulting from a single fresh blastocyst ET in live birth (r = .36, p = .02; r = .31, p = .04, respectively). CONCLUSION: Our study suggests that serum ß-hCG levels resulting from a single fresh blastocyst ET showed a correlation with day 5 blastocoele expansion and blastocyst quality score in both ongoing pregnancy and live birth.


Assuntos
Gonadotropina Coriônica Humana Subunidade beta/sangue , Fase de Clivagem do Zigoto/metabolismo , Fertilização In Vitro/métodos , Transferência de Embrião Único/métodos , Adulto , Biomarcadores/sangue , Feminino , Humanos , Infertilidade Feminina/terapia , Nascimento Vivo , Valor Preditivo dos Testes , Gravidez , Curva ROC , Estudos Retrospectivos
17.
Reprod Biol ; 18(4): 355-360, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30389297

RESUMO

Prolonged embryo culture is increasingly used as a way of improving pregnancy rates, especially in the context of single embryo transfer. So far, only a handful of studies examined the relation between implantation potential and time-lapse parameters extracted from later stages (morula and blastocyst) of embryo development. For this retrospective study all 285 single vitrified-thawed blastocyst transfers (SVBT) from all consecutive unselected patients whose fertilized oocytes were submitted to time-lapse monitoring (TLM) from a two-year cohort were analysed. Two different statistical models were created; a hierarchical one including the two strongest live birth (LB) predictors (t2 and texpB2) and a more complex model based on principal component analysis (PCA) and logistic regression methods. The first, four-category, hierarchical model effectively distinguished between blastocysts of increasing LB rates (8, 30, 40, 53%). For the second data-mining model quartiles of the created Sc parameter had increasing LB rates (12, 19, 40, 49%). AUC values were comparable for both models (0.723, 95CI%:0.66-0.79 versus 0.717, 95CI%:0.65-0.78). The combination of cleavage- and blastocyst-stage variables through hierarchical or data mining-based algorithms was used successfully to predict live birth. However, due to the lack of internal / external validation the predictive capacities of this model could differ largely in different datasets.


Assuntos
Blastocisto/fisiologia , Fase de Clivagem do Zigoto/fisiologia , Implantação do Embrião/fisiologia , Desenvolvimento Embrionário/fisiologia , Nascimento Vivo , Mineração de Dados , Técnicas de Cultura Embrionária , Transferência Embrionária , Feminino , Fertilização , Humanos , Modelos Estatísticos , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Imagem com Lapso de Tempo
18.
Fertil Steril ; 110(4): 703-709, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30196967

RESUMO

OBJECTIVE: To determine whether the freeze-all policy ensures a higher efficacy in terms of cumulative live birth rate (CLBR) in comparison with a conventional fresh/frozen embryo transfer (ET) approach in patients with normal ovarian response. DESIGN: Retrospective, matched, multicenter cohort study. SETTING: Private IVF centers. PATIENT(S): This study analyzed 564 completed IVF cycles in which an average of 12-18 oocytes were retrieved. In 435 cycles the conventional strategy was applied, with initial ET followed by frozen embryo replacements, whereas in 129 cycles the freeze-all policy was performed, with elective cryopreservation and deferred use of all viable embryos. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The primary endpoint was CLBR. The secondary endpoint was cumulative clinical pregnancy rate. RESULT(S): Overall, statistically comparable CLBRs were achieved in the fresh/frozen and freeze-all groups (45.5% vs. 53.5%). Stratification of data for age and number of retrieved oocytes confirmed the absence of differences between the two groups. In a subanalysis in which the day of ET and cryopreservation were taken into account, a similar outcome was achieved in cleavage-stage groups (45.6% vs. 46.4%), whereas when ET was performed at the blastocyst stage the CLBR was significantly higher in the freeze-all group (45.3% vs. 66.7%). CONCLUSION(S): Our CLBR analysis indicates that clinical performance of the freeze-all policy is equivalent to that of the conventional strategy when ET is carried out at the cleavage stage. However, it seems to be superior if associated with cryopreservation and transfer at the blastocyst stage.


Assuntos
Coeficiente de Natalidade/tendências , Blastocisto/fisiologia , Fase de Clivagem do Zigoto/fisiologia , Criopreservação/métodos , Transferência Embrionária/métodos , Adulto , Blastocisto/citologia , Estudos de Coortes , Criopreservação/normas , Transferência Embrionária/normas , Feminino , Humanos , Recuperação de Oócitos/métodos , Recuperação de Oócitos/normas , Indução da Ovulação/métodos , Indução da Ovulação/normas , Estudos Retrospectivos
19.
FEBS Lett ; 592(14): 2414-2424, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29963710

RESUMO

The chromatin structure in one-cell-stage mouse embryos is extremely loose and becomes tighter at the two-cell stage. As linker histones are involved in higher-order chromatin structure, we examined the involvement of the linker histone variant H1foo in the change of chromatin looseness between the one- and two-cell stages. H1foo knockdown causes the chromatin structure to be tighter in the pronucleus and increases deposition of the histone H3 variant H3.1/3.2 in the peripheral region of the pronucleus in one-cell-stage embryos. The decrease in chromatin looseness at the two-cell stage is less after overexpressing H1foo. These results suggest that H1foo is involved in the change in chromatin structure via nuclear deposition of H3 variants between the one- and two-cell stages.


Assuntos
Montagem e Desmontagem da Cromatina/genética , Cromatina/química , Histonas/fisiologia , Zigoto/metabolismo , Animais , Núcleo Celular/metabolismo , Células Cultivadas , Cromatina/metabolismo , Fase de Clivagem do Zigoto/fisiologia , Técnicas de Cultura Embrionária , Feminino , Histonas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Conformação de Ácido Nucleico , Oócitos/metabolismo , Gravidez
20.
PLoS One ; 13(7): e0200273, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29990330

RESUMO

Sperm samples used on fertilization strongly influence the in vitro production (IVP) rates. However, sperm traits behind this effect are not stated consistently until now. This study aimed to evaluate the isolated and combined effect of some sperm traits (MB: total motility before Percoll® gradient, MA: total motility after Percoll® gradient, AI: acrosome integrity, MI: membrane integrity, MP: mitochondrial membrane potential, and CR: chromatin resistance) on IVP rates. This is the first study focusing on the isolated effect of distinct traits. For this purpose, the experiment was divided in three steps. In first step, to study behavior of traits sperm samples (n = 63 batches) were analyzed and ranked based on each trait. In second step, samples ranked were selected from target ranks regions and allocated in groups of four to five batches, creating Higher and Lower groups, according to two different approaches. One aimed to form groups that differed to all sperm traits simultaneously (effect of combined traits). The other aimed to form groups that differed only to a single sperm trait while no differences were observed for the remaining traits (effect of each isolated trait). In third step, for each group successfully formed in step 2, sperm samples were individually and prospectively used for IVP. Cleavage, embryo development and blastocyst rates were recorded and compared between Higher and Lower of respective trait groups. Surprisingly, evaluation of isolated effects revealed that lower levels of MB, AI and MP resulted in higher embryo development and blastocyst rates (p<0.05), which was not observed on cleavage rate. We conclude that sperm traits strongly influence embryo development after in vitro fertilization (IVF), affecting the zygote competence to achieve blastocyst stage. Individually, levels of MB, AI or MP could be some of the key traits that may define IVP efficiency on current systems of embryo production.


Assuntos
Bovinos/embriologia , Espermatozoides/fisiologia , Acrossomo/fisiologia , Animais , Blastocisto/fisiologia , Cromatina/metabolismo , Fase de Clivagem do Zigoto/fisiologia , Técnicas In Vitro , Masculino , Potencial da Membrana Mitocondrial , Povidona , Dióxido de Silício , Motilidade Espermática , Zigoto/fisiologia
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