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Leukemic stem cells
Vobořil, Matouš ; Stöckbauer, Petr (advisor) ; Šídová, Monika (referee)
The biology of stem cells came to the foreground not only due to scientific but also due to clinical interest and it is one of the most developing fields of current biomedical research. The idea, that all tumor cells contain population of cells like stem cells, leads to the "cancer stem cells hypothesis". It says, that each tumor cell contains small population of cells capable to initiate and maintain the tumor growth. Tumors of the hematopoietic tissue were the first, where cancer or leukemic stem cells were isolated. Therefore, leukemic stem cells are so far the best understood cancer stem cells. However, despite the huge advances in the biology of leukemic stem cells, there are many properties still unknown. This thesis initially presents basic knowledges in the stem cell biology, including their origin and identification. Later it focuses on the stem cell hypothesis and describes of the main properties of stem cells in solid tumors. The main part of this thesis also shows in details the origin and properties of the leukemic stem cells and decribes some new directions in the targeted therapy of hematological malignancies.
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Leukemic stem cells
Vobořil, Matouš ; Stöckbauer, Petr (advisor) ; Šídová, Monika (referee)
The biology of stem cells came to the foreground not only due to scientific but also due to clinical interest and it is one of the most developing fields of current biomedical research. The idea, that all tumor cells contain population of cells like stem cells, leads to the "cancer stem cells hypothesis". It says, that each tumor cell contains small population of cells capable to initiate and maintain the tumor growth. Tumors of the hematopoietic tissue were the first, where cancer or leukemic stem cells were isolated. Therefore, leukemic stem cells are so far the best understood cancer stem cells. However, despite the huge advances in the biology of leukemic stem cells, there are many properties still unknown. This thesis initially presents basic knowledges in the stem cell biology, including their origin and identification. Later it focuses on the stem cell hypothesis and describes of the main properties of stem cells in solid tumors. The main part of this thesis also shows in details the origin and properties of the leukemic stem cells and decribes some new directions in the targeted therapy of hematological malignancies.
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Transfection of stem cells and other testicular cells of Xenopus tropicalis
Šídová, Monika ; Tlapáková, Tereza (advisor) ; Petr, Jaroslav (referee)
Xenopus tropicalis represents one of the most important model organism used in developmental and cellular biology. Laboratory of Developmental Biology at the Faculty of Science, Charles University in Prague has successfully established a long-term culture of X. tropicalis juvenile testicular cells. Based on expression profiling analysis of selected specific markers (Sox9, WT1, etc) it was proven that the major cell type in this culture is pre-Sertoli cells. Furthermore these pre-Sertoli cells allow a longterm cultivation of the germinal stem cells. By performing a histochemical test for the presence of alkaline phosphatase in the colony of these cells were proven the features of stem cells. In this diploma thesis we focused on optimization of work with the mixed cell culture. In particular we define conditions of dissociation and subsequent separation of a feeder-layer formed by the pre-Sertoli cells. We also attempted to develop suitable conditions for transfection of the germinal cells. With these techniques we will to investigate the functional properties of the germinal stem cells. Moreover, it provides us a powerful tool for performing another experiments focused on transgenesis and/or different gene inactivation. Powered by TCPDF (www.tcpdf.org)
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Formation of spatio-temporal molecular gradients in early embryonic development of Xenopus laevis.
Šídová, Monika ; Tlapáková, Tereza (advisor) ; Pěknicová, Jana (referee) ; Petr, Jaroslav (referee)
Clarifying the underlying spatio-temporal mechanisms that determine body pattern is important for detailed understanding of embryonic development. A crucial question of vertebrate embryogenesis remains: when and how are single blastomeres determined for differentiation that subsequently leads to body axes specification and the formation of different tissues and organs? The answer to this question will be beneficial for primary research as well as in the field of applied medicine. The main aim of the presented thesis was to study spatio-temporal molecular gradients of cell fate determinants during early embryonic development. The African clawed frog Xenopus laevis was used as a model organism because of their large size of oocytes and external embryonic development. Due to late activation of embryonic transcription, a crucial mechanism of early blastomeres determination is dependent on asymmetric localization of maternal factors within oocyte and their uneven distribution into single blastomeres during early cell division. Two main localization patterns were identified along the animal-vegetal axis of the mature Xenopus oocyte using qPCR tomography. The localization gradient with preference in either animal or vegetal hemisphere was found for maternal mRNA as well as miRNAs. Moreover, two vegetal...
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Single cell expression analysis of genes with potential mrna gradient in mouse oocytes
Dorosh, Andriy ; Margaryan, Hasmik ; Vodička, Martin ; Ergang, Peter ; Šídová, Monika ; Dvořáková-Hortová, Kateřina
In frogs, there are clearly visible differently pigmented animal and vegetal poles of the egg determined before fertilization and leading to asymmetrical divisions. Mammalian egg does not show any comparable differentiation and it has been generally accepted that even the individual blastomeres in 2-cell and 4-cell embryos are homogenous. However, recent findings suggest that those blastomeres display different gene expression patterns and might already possess some inclinations to specific cell lineages. We therefore raised a question, whether there could be any mRNA or protein gradients in pre-fertilization oocytes similar to a previously described amphibian egg one. In mammalian eggs, there is a membrane region that is poor in microvilli, cortical granules are absent beneath plasma membrane and sperm cells generally do not bind to this location. This microvilli free region also covers the egg nucleus, and cytoskeleton localization differs markedly to the rest of the cortical space, forming actin –myosin II cortical cap/ring and is considered as animal pole. The purpose of this study was to determine gene products that can be detected at single cell level using qPCR and display gradient like distribution in mature oocytes. We checked expression of 12 selected genes in a pool of 10 oocytes and single mature oocytes. Then, we analysed gene expression in fixed intact oocytes and those undergoing laser capture microdissection procedure (LCMD). Eventually, we have determined six candidate genes for the study of intracellular spatial gene expression in mature mammalian oocytes by subcellular qPCR and in situ hybridization.
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