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Comparison of gene expression in fresh and frozen–thawed human preimplantation embryos

机译:新鲜和冻融的人类植入前胚胎中基因表达的比较

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Identification and characterisation of differentially regulated genes in preimplantation human embryonic development are required to improve embryo quality and pregnancy rates in IVF. In this study, we examined expression of a number of genes known to be critical for early development and compared expression profiles in individual preimplantation human embryos to establish any differences in gene expression in fresh compared to frozen–thawed embryos used routinely in IVF. We analysed expression of 19 genes by cDNA amplification followed by quantitative real-time PCR in a panel of 44 fresh and frozen–thawed human preimplantation embryos. Fresh embryos were obtained from surplus early cleavage stage embryos and frozen–thawed embryos from cryopreserved 2PN embryos. Our aim was to determine differences in gene expression between fresh and frozen–thawed human embryos, but we also identified differences in developmental expression patterns for particular genes. We show that overall gene expression among embryos of the same stage is highly variable and our results indicate that expression levels between groups did differ and differences in expression of individual genes was detected. Our results show that gene expression from frozen–thawed embryos is more consistent when compared with fresh, suggesting that cryopreserved embryos may represent a reliable source for studying the molecular events underpinning early human embryo development.
机译:在植入前人类胚胎发育中鉴定和鉴定差异调节基因是提高IVF胚胎质量和妊娠率所必需的。在这项研究中,我们检查了许多已知对早期发育至关重要的基因的表达,并比较了单个植入前人类胚胎中的表达谱,以建立与IVF中常规使用的冻融胚胎相比,新鲜胚胎中基因表达的任何差异。我们通过cDNA扩增和定量实时PCR分析了一组44个新鲜和冻融的人类植入前胚胎中19个基因的表达。新鲜的胚胎是从剩余的早期卵裂期胚胎中获得的,而冷冻融化的胚胎则是从2PN冷冻保存的胚胎中获得的。我们的目的是确定新鲜和冰冻融化的人类胚胎之间基因表达的差异,但我们也发现了特定基因在发育表达方式上的差异。我们表明,同一时期胚胎之间的整体基因表达是高度可变的,我们的结果表明,各组之间的表达水平确实存在差异,并且检测到各个基因表达的差异。我们的结果表明,与新鲜相比,冻融胚胎的基因表达更为一致,这表明冷冻保存的胚胎可能是研究支撑人类早期胚胎发展的分子事件的可靠来源。

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