235 resultados para BLASTOCYST


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Pós-graduação em Medicina Veterinária - FMVZ

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This study evaluated the influence of follicular fluid (FF) added to the maturation medium on the quality of bovine embryos produced in vitro. In the first experiment, oocytes were matured in media containing five different FF concentrations with different maturation times and classified according to meiotic progression and migration of cortical granules. In the second experiment, oocytes matured in the same media were fertilized at three different maturation times; thereafter, cleavage and blastocyst rates were evaluated. In the third experiment, oocytes were matured in media containing three different FF concentrations at two different maturation times, and embryo quality, inferred by the ratio of inner cell mass and trophectoderm cells compared with total cell number, was evaluated. Higher FF concentration (75 - 100% FF) slowed meiotic progression and CG migration (control - 78.13% vs. treated - 52.58% and control - 52.7% vs. treated - 11.59%, respectively, at 24 h of maturation). Also, FF at concentration of 75% or 100% had a negative influence on cleavage and blastocyst rates (control - 90.13% vs. treated - 82.64% and control - 35.73% vs. treated - 11.57%, respectively, at 24 h of maturation). The 50% FF resulted in embryos with increased inner cell mass numbers (control - 29.91 vs. treated - 35.49, at 24 h of maturation) and total cell numbers (control - 109.53 vs. treated - 120.67, at 26 h of maturation). Even though higher concentration of FF added to the maturation medium reduced embryonic development rates, in lower concentrations, FF slowed the meiotic progression and migration of CG and contributed to increases in inner cell mass number. Thus, FF added to the maturation medium enhances the number of cells in bovine embryos produced in vitro, especially for inner cell mass.

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Since cyclooxygenase (COX) inhibitors have been pointed out as potential treatments to increase pregnancy rates after embryo transfer, the present experiment aimed to evaluate the effects of flunixin meglumine (FM) and parecoxib (P), a COX-1 and 2 or COX-2 specific inhibitor, respectively, on the development of bovine embryos until the hatched blastocyst stage. In vitro produced bovine embryos were cultured in media with different concentrations of FM (0.14; 1.4; 14; 140 or 1400 mu g/ml) or P (0.09; 0.9; 9; 90 or 900 mu g/ml) and the production rates were evaluated. Concentrations of FM <= 14 mu g/ml and P <= 90 mu g/ml did not impair embryo development, although compiled data from non-lethal FM concentrations (<= 14 mu g/ml) indicated a toxic effect enough to decrease the hatching rate of blastocysts. Concentrations of FM at 140 and 1400 mu g/ml and P at 900 mu g/ml were lethal as no cleavage was detected on presumptive zygotes.

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The Brazilian livestock stands out for having the world largest commercial herd of cattle and leads meat exportation and production of bovine embryos. The in vitro production (IVP) of embryos is considered an effective option to overcome problems such as infertility in cows with high economic value and also for genetic improvement of cattle. The in vitro oocyte maturation is an essential step to the success of IVP, but is still considered poor when compared to in vivo maturation. Recent studies have suggeested an important role of Fibroblast Growth Factor 10 (FGF10) on the in vitro maturation of oocytes, which favored the expression of genes related to oocyte maturation and cumulus cell expansion. Aware that maturity stage influences the final production of blastocysts, we aimed study to verify if the addition of FGF10 into the maturation medium is able to affect positively the IVP of bovine embryos. Hence, FGF10 was added to maturation in five different concentrations: 0.5 ng/mL (group 0.5), 2.5 ng/mL (group 2.5), 5 ng/mL (group 5), 10 ng/mL (group 10) and 50 ng/mL (group 50). Additionally, two other maturation groups were used, group BSA (Bovine Serum Albumin, 4 mg/mL) and group FCS (Fetal Calf Serum, 10%). The rates of cleavage, morula and blastocyst were analyzed by Analysis of Variance (ANOVA), differences of P<0.05 were considered significant. Cleavage rates did not differ between the seven groups. On the other hand, morula rate on FCS group was higher than groups BSA, 0.5, 10 and 50 (P<0.05), but did not differ among groups treated with intermediate doses of FGF10 (2.5 and 5). FCS group presented higher blastocyst rate compared to all other groups that were well below the FCS group (P<0.0001). Therefore, the use of FGF10 during oocyte maturation did not affect positively embryo development on the IVP of bovine embryos

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In vitro production has been employed in bovine embryos and quantification of lipids is fundamental to understand the metabolism of these embryos. This paper presents a unsupervised segmentation method for histological images of bovine embryos. In this method, the anisotropic filter was used in the differents RGB components. After pre-processing step, the thresholding technique based on maximum entropy was applied to separate lipid droplets in the histological slides in different stages: early cleavage, morula and blastocyst. In the postprocessing step, false positives are removed using the connected components technique that identify regions with excess of dye near pellucid zone. The proposed segmentation method was applied in 30 histological images of bovine embryos. Experiments were performed with the images and statistical measures of sensitivity, specificity and accuracy were calculated based on reference images (gold standard). The value of accuracy of the proposed method was 96% with standard deviation of 3%.

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