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Genome-wide profiling of H1 linker histone variants in mouse embryonic stem cellsCao, Kaixiang 22 May 2014 (has links)
H1 linker histone facilitates the formation of higher order chromatin structure and is essential for mammalian development. Mice have 11 H1 variants which are differentially regulated and conserved in human. Previous research indicates that H1 regulates the expression of specific genes in mouse embryonic stem cells (ESCs). However, whether individual variants have distinct functions and how H1 participates in gene regulation remain elusive. An investigation of the precise localization of individual H1 variants in vivo would facilitate the elucidation of mechanisms underlying chromatin compaction regulated gene expression, while it has been extremely difficult due to the lacking of specific antibodies toward H1 variants. In this dissertation, I have generated a knock-in system in ESCs and shown that the N-terminally tagged H1 proteins are functionally interchangeable to their endogenous counterparts in vivo. H1d and H1c are depleted from GC- and gene-rich regions and active promoters, inversely correlated with H3K4me3, but positively correlated with H3K9me3 and associated with characteristic sequence features. Surprisingly, both H1d and H1c are significantly enriched at major satellites, which display increased nucleosome spacing compared with bulk chromatin. While also depleted at active promoters and enriched at major satellites, overexpressed H10 displays differential binding patterns in specific repetitive sequences compared with H1d and H1c. Depletion of H1c, H1d ,and H1e causes pericentric chromocenter clustering and de-repression of major satellites. Collectively, these results integrate the localization of an understudied type of chromatin proteins, namely the H1 variants, into the epigenome map of mouse ESCs, and demonstrate significant changes at pericentric heterochromatin upon depletion of this epigenetic mark.
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Characterization of ceramide synthases (Cers) in mammalian cellsPark, Hyejung 13 May 2009 (has links)
This thesis describes the characterization of ceramide (Cer) biosynthesis by mammalian cells. The possibility that Cer undergo developmental changes was explored using mouse embryonic stem cells versus embryoid bodies by analysis of the Cer subspecies by liquid chromatography, electrospray ionization-tandem mass spectrometry (LC ESI-MS/MS) and of the transcript levels for enzymes involved in Cer biosynthesis by qRT-PCR. Cer of embroid bodies had higher proportions of very-long-chain fatty acids, which correlated with the relative expression of mRNA for the respective Cer synthases (CerS) and fatty acyl-CoA elongases, as well as changes in the fatty acyl-CoA's of the cells. Therefore, it is clear that Cer subspecies change during embryogenesis, possibly for functionally important reasons. One CerS isoform, CerS2, was studied further because it has the broadest tissue distribution and a remarkable fatty acyl-CoA specificity, utilizing longer acyl-chain CoAs (C20-C26) in vitro. The fatty acid chain selectivity was refined by analysis of the Cer from livers from CerS2 null mice, which displayed very little Cer with fatty acyl chains with 24 + 2 carbons. Another interesting structural variation was discovered in studies of cells treated with fumonisin B1 (FB1), which inhibits CerS. Under these conditions, cells in culture and animals accumulate substantial amounts of a novel sphingoid base that was identified as 1-deoxysphinganine. This compound arises from utilization of L-alanine instead of L-serine by serine palmitoyltransferase (SPT) based on the inability of LYB cells, which lack SPT, to make 1-deoxysphinganine. In the absence of FB1, 1-deoxysphinganine is primarily acylated to 1-deoxydihydroceramides. These are an underappreciated category of bioactive sphingoid bases and "ceramides" that might play important roles in cell regulation and disease. In summary, cells contain a wide variety of Cer subspecies that are determined by changes in expression of CerS, enzymes that produce co-substrates (such as fatty acyl-CoAs), and the types of amino acids utilized by SPT, the initial enzyme of de novo sphingolipid biosynthesis. One can envision how these changes might impact membranes structure as well as signaling by this family of highly bioactive compounds.
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Characterisation of human fetal mesenchymal stem cells /Götherström, Cecilia, January 2004 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.
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Cell damage and tissue repair in the central nervous system : electron mi[c]roscopy study of neuronal death and cell replacement /Andersson, Benita, January 2005 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 6 uppsatser.
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Neural stem and progenitor cells cellular responses to known and novel factors /Larsson, Jimmy, January 2010 (has links)
Diss. (sammanfattning) Uppsala : Uppsala universitet, 2010. / Härtill 4 uppsatser.
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Role of membrane-type 1 matrix metalloproteinase in hematopoietic stem/progenitor cell traffickingShirvaikar, Neeta Chandan. January 2010 (has links)
Thesis (Ph.D.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Medicine. Title from pdf file main screen (viewed on April 27, 2010). Includes bibliographical references.
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Stem κύτταρα και μικροπεριβάλλον στον καρκίνο των ωοθηκώνΒίτσας, Χαράλαμπος 29 July 2011 (has links)
Τα stem κύτταρα είναι ένας υποπληθυσμός κυττάρων με δύο κύριες ιδιότητες: αυτοανανέωση και διαφοροποίηση. Τα stem κύτταρα διαμένουν σε ένα εξειδικευμένο μικροπεριβάλλον, την φωλεά, η οποία παίζει σημαντικό ρόλο στη διατήρηση της ισορροπίας μεταξύ της αυτοανανέωσης και της διαφοροποίησης. Τελευταία δεδομένα εισηγούνται ότι ο καρκίνος αναπτύσσεται από ένα υποσύνολο κυττάρων με ιδιότητες ανάλογες αυτών των φυσιολογικών stem κυττάρων. Τα κύτταρα αυτά αποκαλούνται καρκινικά stem κύτταρα. Η θεωρία των καρκίνικών stem κυττάρων υποστηρίζει ότι τα καρκινικά stem κύτταρα εγκαινιάζουν και συντηρούν την ανάπτυξη και εξέλιξη του όγκου, ευθύνονται για την κυτταρική ετερογένεια των καρκίνικών κυττάρων του όγκου, είναι υπεύθυνα για τις μεταστάσεις και παραμένουν στους ασθενείς παρά τη χρήση των συμβατικών χημειοθεραπευτικών παραγόντων. Πρόσφατα δεδομένα πιστοποιούν την ύπαρξη καρκινικών stem κυττάρων στην ωοθήκη. / Stem cells are a subpopulation of cells with two key properties: self-renewal and differentiation. Stem cells reside in a specialized microenvironment, i.e. niche, which plays an important role in the balance between self-renewal and differentiation. Recent data suggest that cancer develops from a subset of cells with properties similar to those of normal stem cells.
These cells are called cancer stem cells. Cancer stem cell hypothesis suggest that cancer stem cells initiate and preserve the growth of tumor, they are responsible for cellular heterogeneity and metastasis of tumor and they are, finally, drug-resistant.Latest data suggest the presence of cancer stem cells in the ovary.
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Establishment in culture of mouse and human stem cells with expanded fate potentialRyan, David John January 2018 (has links)
The zygote and blastomeres of cleavage stage mouse embryos have the capacity to differentiate to the embryonic and both extra-embryonic lineages and are considered functionally totipotent. Until now, it has not been possible to establish stable cell lines that resemble these totipotent-like cells. In this work, I hypothesised that by modulating signalling pathways known to be important in early embryonic development it may be possible to capture in vitro a self-renewing state that possessed features of pre-implantation blastomeres. I succeeded in formulating a novel hypothesis driven cell culture medium which allowed the establishment of a stem cell state that possessed expanded differentiation potential to the embryonic and both extra-embryonic lineages in vitro and in vivo. These cells were isolated directly from in vitro culture of mouse pre-implantation embryos or single cell blastomeres, reprogrammed from somatic cells or converted from mouse ES cells. With these cells, I generated single cell chimeras which demonstrated extensive contribution to all lineages in the developing organism providing additional evidence that this chemical medium maintained a homogenous stem cell population. I demonstrated that the transcriptome of these cells was enriched with an early pre-implantation blastomere signature, distinct from other rare published totipotent-like cells. Finally, I demonstrated that the same chemical formulation permitted the establishment in vitro of a human stem cell state that possessed expanded differentiation potential to the embryonic and extra-embryonic lineage in vitro. My work has shown for the first time that through chemical modulation of pathways implicated to be involved in pre-implantation development, a novel homogenous stem cell state that possesses a pre-implantation transcriptional signature and expanded differentiation potential to both the embryonic and extra-embryonic lineage can be established and maintained in vitro in both mouse and human, suggesting a possible interspecies conservation of the signalling networks involved in early embryonic development.
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The role of human embryonic stem cell-derived epicardium in myocardial graft developmentBargehr, Johannes January 2018 (has links)
No description available.
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A History of Cellular Senescence and Its Relation to Stem Cells in the Twentieth and Twenty-First CenturiesJanuary 2015 (has links)
abstract: Researchers in the twentieth and twenty-first centuries identify the study of the intrinsic and external factors that influence human aging as senescence. A commonly held belief in the year 2015 is that at least some kinds of cells can replicate over long periods or even indefinitely, thereby meaning the cell does not undergo senescence (also known as replicative senescence) and is considered immortal. This study aims to provide information to answer the following question: While some scientists claim they can indefinitely culture a stem cell line in vitro, what are the consequences of those culturing practices? An analysis of a cluster of articles from the Embryo Project Encyclopedia provides information to suggest possible solutions to some potential problems in cell culturing, recognition of benefits for existing or historical culturing practices, and identification of gaps in scientific knowledge that warrant further research. Recent research suggests that hESCs, and immortalized cell lines in general, do not escape the effects of senescence. While there exists a constant change in the practices of cell culturing, a large portion of scientists still rely on practices established before modern senescence research: research that seems to suggest that cultured hESCs, among other immortal cell lines, are not truly immortal. / Dissertation/Thesis / Masters Thesis Biology 2015
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