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Pregnancy derived from human zygote pronuclear transfer in a patient who had arrested embryos after IVF
Nuclear transfer of an oocyte into the cytoplasm of another enucleated oocyte has shown that embryogenesis and implantation are influenced by cytoplasmic factors. We report a case of a 30-year-old nulligravida woman who had two failed IVF cycles characterized by all her embryos arresting at the two-cell stage and ultimately had pronuclear transfer using donor oocytes. After her third IVF cycle, eight out of 12 patient oocytes and 12 out of 15 donor oocytes were fertilized. The patient's pronuclei were transferred subzonally into an enucleated donor cytoplasm resulting in seven reconstructed zygotes.System for evaluation of oxidative stress on in-vitro-produced bovine embryos
This study proposed a quantitative evaluation of oxidative status (OS) in bovine embryos. Sixteen-cell stage embryos, cultured under 5% O2, were treated with oxidative stress inducer menadione (0, 1, 2.5 and 5 µmol/l) for 24 h. Blastocyst rate (BLR) was recorded and expanded blastocysts were stained with CellROX®Green (CRG; OS evaluation) and evaluated under epifluorescence microscopy (ratio of pixel/blastomere). A significant effect of menadione was observed for BLR (P = 0.0039), number of blastomeres/embryo (P < 0.0001) and OS (P < 0.001).Cytoplasmic, rather than nuclear-DNA, insufficiencies as the major cause of poor competence of vitrified oocytes
An oocyte is a unique body cell that is developmentally committed to support fertilization and early embryonic development (Hosseini et al., 2012). This capacity strongly depends on the cellular and molecular aspects of nucleo-cytoplasmic interactions, which facilitate early mitotic divisions of the embryo until broad embryonic genome activation (Sirard, 2012). During vitrification, this highly organized structure often incurs serious damage which inevitably affects its capacity to respond to subsequent treatment (Smith et al., 2011).Stability of AMH measurement in blood and avoidance of proteolytic changes
The new Gen II assay for anti-Müllerian hormone (AMH) shows good stability and reliability in serum, but analyses of stability in whole blood are lacking. Testing the effects of storage of whole-blood samples at room temperature revealed significant increases in the measured value of AMH of 31% over 4 days (P < 0.001). The effect is temperature dependent, with storage at 4°C showing markedly reduced increments. Further, samples collected into serum tubes with gel separators and centrifuged within 5 h (blood cells and serum physically separated within the collection tube) showed reliable stability over a period of more than 5 days.
