ABSTRACT
The mammalian zygote cleaves and develops to blastocyst within the zona pellucida (ZP) in vivo. The presence or absence of ZP may affect the characteristics of the embryo, including blastomere alignment, cell-cell junction, and compaction. This study aimed to compare the morphokinetic characteristics of ZP intact and ZP free mouse pre-implantation embryos with time-lapse monitoring system. Mouse 2-cell embryos were collected 1.5 days post coitum (dpc), and their ZPs were removed by treatment with acid Tyrode's solution. All embryos were cultured in vitro up to the outgrowth stage at 7.5 dpc. In this study, ZP did not influence the cumulative times from 2-cell to further stages and blastulation. Interestingly, ZP free embryos at 4-cell stage have three patterns of blastomere alignment according to the number of contact points between blastomeres. However, blastomere alignment did not lead to any differences in morphokinetic comparisons. Regardless of the presence or absence of ZP, embryos compacted after the 8-cell stage took shorter time to become blastocysts than embryos compacted pre-8-cell stage. Nevertheless, cell-cell junction proteins required for successful compaction were similarly expressed between ZP intact and ZP free embryos. ZP intact embryos compacted post-8-cell stage had higher rate of reaching blastocysts than compacted ZP intact embryos before 8-cell stage while the outgrowth/blastocyst rate was similar. In this study, the presence or absence of ZP did not influence embryonic development and expression of cell surface glycoproteins, whereas compaction timing may be one of the criteria for evaluating embryo quality. ZP free embryos may become an alternative for overcoming cases with ZP problems in a human ART program.
Subject(s)
Embryo, Mammalian/embryology , Embryonic Development , Microscopy, Fluorescence/methods , Time-Lapse Imaging/methods , Zona Pellucida/metabolism , Animals , Blastocyst/cytology , Blastocyst/metabolism , Blastomeres/cytology , Blastomeres/metabolism , Cadherins/metabolism , Embryo Culture Techniques/methods , Embryo, Mammalian/cytology , Embryo, Mammalian/metabolism , Female , Male , Mice, Inbred ICR , beta Catenin/metabolismABSTRACT
Recently, many studies have investigated the role of extracellular vesicles (EVs) on reproductive events, including embryo development and death, oviduct-embryo crosstalk, in vitro fertilization and others. The aim of this study was to demonstrate whether outgrowth embryo-derived EVs function as bioactive molecules and regulate mouse embryonic developmental competence in vitro and implantation potential in utero. The EVs from mouse outgrowth embryos on 7.5 days postcoitum were detected and selectively isolated to evaluate the embryotrophic functions on preimplantation embryos. Developmental outcomes such as the percentage of blastocyst formation, hatching, and trophoblastic outgrowth were assessed. Furthermore, the total cell number and apoptotic index of blastocysts, which were incubated with EVs during the culture period, were evaluated by fluorescence microscopy. Implantation potential in utero was investigated following embryo transfer. The EVs from outgrowth embryo-conditioned media have rounded membrane structures that range in diameter from 20 to 225 nm. Incubation with EVs improved preimplantation embryonic development by increasing cell proliferation and decreasing apoptosis in blastocysts. Moreover, the implantation rates following embryo transfer were significantly higher in EV-supplemented embryos compared with the control. Collectively, EVs from outgrowth embryo could enhance the embryonic developmental competence and even implantation potential in mice.
Subject(s)
Blastocyst/metabolism , Cell Proliferation , Embryonic Development , Extracellular Vesicles/transplantation , Animals , Blastocyst/cytology , Culture Media, Conditioned/pharmacology , Embryo Culture Techniques , Female , Mice , Mice, Inbred ICRABSTRACT
Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free fatty acid-induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with palmitic acid (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free fatty acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.
