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Molecular mechanisms regulating participation of neural crest on developmental processes and homeostasis in selected localizations
Krejčí, Eliška ; Grim, Miloš (advisor) ; Hampl, Aleš (referee) ; Jendelová, Pavla (referee)
This PhD thesis focuses on the study of neural crest (NC) properties and mechanisms regulating its development. It is based on four original papers, two of which are wholy concerning NC. In the first project we showed that transcription factor C-MYB is expressed in majority of cells in the developing chick embryo. Its expression level is elevated in neural folds and migrating NC cells. Other experiments showed that this elevated level of intracellular c-Myb influences induction and epithelial-mesenchymal transformation of NC cells, and that c-Myb lies within the BMP4 signalling cascade. In the second project we explored the spatio-temporal specification along the antero-posterior axis of the limb bud - a mechanism significantly regulating also NC development. We showed that deletion of regulatory element of Plzf transcription repressor gene caused decrease of Plzf expression in the limb buds followed by anterior expansion of its downstream genes (Hoxd10-13) expression domains only in the hind limb, with a resulting additional finger. The third paper studies developmental origin of medial border of mammalian scapula. We showed that its cells originate from somites and not in NC. In the last project we studied NC stem cells isolated from human adult hair follicles. These cells express NC stem cell...
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Primary Cilia and its Importance in Cell Response to Ionizing Radiation Exposure and Chemotherapy Drugs.
Filipová, Alžběta ; Řezáčová, Martina (advisor) ; Hampl, Aleš (referee) ; Hofer, Michal (referee)
Primary cilia and its importance in cell response to ionizing radiation exposure and chemotherapy drugs. Ionizing radiation, metabolic and genotoxic stress affect the cytoskeletal stability and morphology of cells by causing DNA damage in the form of single or double stranded breaks, the latter being the most critical. When DNA damage occurs the cell cycle becomes arrested and the repair machinery is engaged; however, if this damage is not repaired the cell enters programmed cell death by apoptosis. Primary cilia have been shown to act as physical- chemical sensors and their biological functions include the perception of the extracellular milieu, the regulation of organogenesis and cell polarity, as such, they are dynamically regulated during cell cycle progression. Further, the impairment or loss of primary cilia leads to the development of ciliopathies and other diseases including cardiovascular disorders, arthritis and, ultimately, cancer. Moreover, the presence and temporary formation of primary cilia is essential for the repair process of certain cell types. This work is focused on the evaluation of primary cilia changes caused by ionizing radiation, metabolic stress or cytostatic drugs (taxol and doxorubicin) in vitro. Our results showed a significantly higher number of ciliated C2C12 cells observed...
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Dental pulp stem cells
Suchánek, Jakub ; Koberová Ivančaková, Romana (advisor) ; Kukletová, Martina (referee) ; Hampl, Aleš (referee)
The aim of this dissertation study was to optimize the isolation and long term cultivation protocols for human dental pulp stem cells. The protocols which showed best results were used for cultivation of dental pulp stem cell isolated from exfoliated teeth (SHED). Additional aims were to characterize DPSC and SHED and prove their ability to proliferate over Hayflick's limit and differentiate into mature cell lines (osteoblasts, chondroblasts and adipocytes). In order to find optimal protocols for isolation of dental pulp from tooth, we tried three different approaches. During optimization of cultivation protocol we focused on decreasing amount of fetal calf serum (FCS) from 10 % FCS in cultivation media (most often used in literature) into 2 % and thus get closer to cultivation conditions suitable for clinical usage. We compared DPSC cultivated in three different media (medium with 10 % FCS, 2 % FCS supplemented with growth factors and media with 2 % FCS supplemented with ITS and growth factors). For characterization of DPSC and SHED we used basic biological properties (proliferation activity, viability, morphology), their phenotype and karyotype. The study demonstrated that the best protocol for isolation of dental pulp from tooth was to break the roots and extract the pulp throw this aperture. We...
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Cytokine expression in chemically-induced senescence
Nováková, Zora ; Hodný, Zdeněk (advisor) ; Hampl, Aleš (referee) ; Dvořák, Michal (referee)
AbstractJanderova-Rossmeislova L., Novakova Z., Vlasakova J., Philimonenko V., Hozak P., Hodny Z., 2007. PML protein association with specific nucleolar structures differs in normal, tumor and senescent human cells. J. Struct. Biol. 159, 56-70. Cellular senescence, widely recognized as a potent suppressor of tumorigenesis, represents response to cellular stress and DNA damage which results in irreversible cell growth arrest. Growing evidence signalizes crucial role of cytokine production in senescence phenomenon. Recently, many research groups have efforted to define exact character of senescence-associated secretory phenotype and its function in senescence development and maintentance. Factors secreted by senescent cells, mainly of proiflammatory character, were found to have pronounced effects on their environment as well as on its own producer. These observations were obtained preferentially on models of replicative senescence and oncogene-induced senescence. Vlasakova J, Novakova Z, Rossmeislova L, Kahle M, Hozak P, Hodny Z. 2007. Histone deacetylase inhibitors suppress IFNalpha-induced up-regulation of promyelocytic leukemia protein. Blood 109: 1373-80 Novakova Z, Man P, Novak P, Hozak P, Hodny Z. 2006. Separation of nuclear protein complexes by blue native polyacrylamide gel electrophoresis....
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