36 research outputs found

    A Simple Approach for COnsumption and RElease (CORE) Analysis of Metabolic Activity in Single Mammalian Embryos

    Get PDF
    Non-invasive assay of the consumption and release of metabolites by individual human embryos could allow selection at the cleavage stage of development and facilitate Single Embryo Transfer in clinical IVF but will require simple, high throughput, sensitive methods applicable to small volume samples. A rapid, simple, non-invasive method has therefore been devised using a standard fluorescence plate reader, and used to measure the consumption of pyruvate and glucose, and release of lactate by single bovine embryos at all stages of preimplantation development in culture; amino acid profiles have been determined using HPLC. Early embryos with an ‘intermediate’ level (6.14±0.27 pmol/embryo/h) of pyruvate uptake were associated with the highest rate (68.3%) of blastocyst development indicating that a mid “optimum” range of pyruvate consumption correlates with high viability in this bovine model

    The placenta: phenotypic and epigenetic modifications induced by Assisted Reproductive Technologies throughout pregnancy

    Get PDF

    Analysis of H19 methylation in control and abnormal human embryos, sperm and oocytes

    No full text
    International audienceART is suspected to generate increased imprinting errors in the lineage. Following an ICSI (Intra Cytoplamic Sperm Injection) procedure, a certain number of embryos fail to develop normally and imprinting disorders may be associated to the developmental failure. To evaluate this hypothesis, we analysed the methylation profile of H19DMR, a paternally imprinting control region, in high graded blastocysts, in embryos showing developmental anomalies, in the matching sperm and in oocytes of the concerned couples when they were available. Significant hypomethylation of the paternal allele was observed in half the embryos, independently of the stage at which they were arrested (morula, compacted morula, pre blastocyst or BC graded blastocysts). Conversely, some embryos showed significant methylation on the maternal allele, whereas few others showed both, hypomethylation of the paternal allele and abnormal methylation of the maternal allele. The matching sperm at the origin of the embryos exhibited normal methylated H19 patterns. Thus, hypomethylation of the paternal allele in the embryos does not appear inherited from the sperm but likely reflects instability of the imprint during the demethylating process which occurred in the early embryo. Analysis of a few oocytes suggests that the defect in erasure of the paternal imprint in the maternal germ line may be responsible for the residual methylation of the maternal allele in some embryos. None of these imprinting alterations could be related to a particular stage of developmental arrest; compared to high grade blastocysts, embryos with developmental failure are more likely to have abnormal imprinting at H19 (p<0.05)
    corecore