27 research outputs found

    Variations of X Chromosome Inactivation Occur in Early Passages of Female Human Embryonic Stem Cells

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    X chromosome inactivation (XCI) is a dosage compensation mechanism essential for embryonic development and cell physiology. Human embryonic stem cells (hESCs) derived from inner cell mass (ICM) of blastocyst stage embryos have been used as a model system to understand XCI initiation and maintenance. Previous studies of undifferentiated female hESCs at intermediate passages have shown three possible states of XCI; 1) cells in a pre-XCI state, 2) cells that already exhibit XCI, or 3) cells that never undergo XCI even upon differentiation. In this study, XCI status was assayed in ten female hESC lines between passage 5 and 15 to determine whether XCI variations occur in early passages of hESCs. Our results show that three different states of XCI already exist in the early passages of hESC. In addition, we observe one cell line with skewed XCI and preferential expression of X-linked genes from the paternal allele, while another cell line exhibits random XCI. Skewed XCI in undifferentiated hESCs may be due to clonal selection in culture instead of non-random XCI in ICM cells. We also found that XIST promoter methylation is correlated with silencing of XIST transcripts in early passages of hESCs, even in the pre-XCI state. In conclusion, XCI variations already take place in early passages of hESCs, which may be a consequence of in vitro culture selection during the derivation process. Nevertheless, we cannot rule out the possibility that XCI variations in hESCs may reflect heterogeneous XCI states in ICM cells that stochastically give rise to hESCs

    Embryonic Diapause Is Conserved across Mammals

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    Embryonic diapause (ED) is a temporary arrest of embryo development and is characterized by delayed implantation in the uterus. ED occurs in blastocysts of less than 2% of mammalian species, including the mouse (Mus musculus). If ED were an evolutionarily conserved phenomenon, then it should be inducible in blastocysts of normally non-diapausing mammals, such as domestic species. To prove this hypothesis, we examined whether blastocysts from domestic sheep (Ovis aries) could enter into diapause following their transfer into mouse uteri in which diapause conditions were induced. Sheep blastocysts entered into diapause, as demonstrated by growth arrest, viability maintenance and their ED-specific pattern of gene expression. Seven days after transfer, diapausing ovine blastocysts were able to resume growth in vitro and, after transfer to surrogate ewe recipients, to develop into normal lambs. The finding that non-diapausing ovine embryos can enter into diapause implies that this phenomenon is phylogenetically conserved and not secondarily acquired by embryos of diapausing species. Our study questions the current model of independent evolution of ED in different mammalian orders

    Stříbrem dopovaný hydroxidapatitový povlak nanášený pomocí suspenzního plazmatického nástřiku

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    Čistá suspenze hydroxidapatatu byla vyrobena pomocí chemické sysntézy. S použitím WSP hořáku byl vytvořen HA povlak na substrátu z SS304 a Ti6Al4V. Vlastnosti povlaku byly hodnoceny pomocí SEM, XRD a EDX. PO dopozici bylo dosaženo 4-10% podílu amorfní fáze a 75-82% krystalické HA fáze v tloušťcě cca 145 um. Tribologické chování bylo hodnoceno pomocí testu pin-on-disc. Přídavek AgNO3 do suspenze vedl k vzniku Ag disperze mezi splaty v 8% podílu Ag. Úspěšné přidání antibakteriálního Ag je pokrokem v oblasti výzkumu materiálů kloubních náhrad.Pure hydroxyapatite suspension was produced by wet chemical synthesis. Using a hybrid water-stabilized torch, a series of HA coatings were produced on SS304 and Ti6Al4V substrates and their properties were characterized by SEM, EDX and XRD techniques. After deposition, the amorphous phase content reached 6-10% and the coatings retained 75-82% of crystalline HA phase. Their thickness reached 145 lm. To understand the wear behavior of the coatings, pin-on-disc tribology evaluation was performed. Additionally, a set of HA coatings was prepared with pure metallic Ag content. This formed by in situ chemical decomposition of AgNO3 added into the HA suspension. The Ag was dispersed evenly within the coatings in the form of submicron-sized particles situated predominantly along the HA splats boundaries with a total Ag content of 8 wt.%. Given the antibacterial properties of Ag, such result presents a promising step forward in the hard tissue replacement research
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