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• W2165023364 abstract "In comparison to other mammalian species, relatively little is known regarding the physiology of the human preimplantation embryo, in particular protein production. This has been in part due to the paucity of material and the lack of sensitivity of conventional protein analysis. With recent advances in mass spectrometry it has become possible to analyze the protein production profile into the surrounding medium (secretome) of decreasing numbers of cells. We have therefore developed a system capable of analyzing the secretome of the human preimplantation embryo at each successive stage of development. The goal of this work is to develop a completely non-invasive assay to quantitate human embryo viability facilitating the identification of embryos with the greatest developmental potential. Experimental study. Cryopreserved day 2 and day 3 human embryos were donated with consent for research. Embryos were thawed and subsequently cultured in sequential media (G1/G2) supplemented with recombinant human albumin at 37°C, 5% O2 & 6% CO2. Embryos were moved to fresh 10μl drops of media every 24h and 5μl of spent media were subsequently processed and analyzed by time-of-flight mass spectrometry to determine the embryo secretome at each developmental stage up to the hatching blastocyst. Unique secretome profiles were reliably identified between embryos of different morphologies. Significantly, several unique proteins/biomarkers were observed in the secretome of developing embryos (between 2 to 14 kDa). Data suggests that the protein produced in highest abundance by the human preimplantation embryo was ubiquitin (8.5 kDa). Production of ubiquitin by the cleavage stage embryo did not appear to be related to morphology. However, from the early to the hatching blastocyst stage ubiquitin production was directly associated with morphology. Embryos undergoing degeneration exhibited a unique secretome at all developmental time points. This is the first study to utilize a system involving time-of-flight mass spectrometry to successfully characterize the secretome of the human embryo throughout the preimplantation period. As well as documenting the discovery of several significantly produced proteins/biomarkers the data suggests the identification of a major product of the human embryo to be the protein, ubiquitin. The significance of this finding is that the ubiquitin-proteosome pathway has been implicated to be involved in the implantation process of other mammalian species. This study has also shown that several unique proteins/biomarkers are secreted during the preimplantation period and that degenerating embryos exhibit a significantly different secretome profile. As such a panel of differentially expressed proteins/biomarkers including quantification of ubiquitin production by the human embryo could form the base of a non-invasive assay to determine embryo viability. This approach of protein analysis will further increase our understanding of human embryo physiology and its interaction with the female reproductive tract." @default.
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• W2165023364 date "2005-09-01" @default.
• W2165023364 modified "2023-09-30" @default.
• W2165023364 title "A Novel Non-invasive Means Of Identifying The Protein Production Profile Into The Culture Medium (secretome) Of The Human Preimplantation Embryo" @default.
• W2165023364 doi "https://doi.org/10.1016/j.fertnstert.2005.07.134" @default.
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