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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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Replication of Merkel cell polyomavirus (MCPyV) in human cell lines
Bučková, Alžbeta ; Saláková, Martina (advisor) ; Huerfano Meneses, Sandra (referee)
The Merkel cell polyomavirus (MCPyV) is the only human polyomavirus classified as probably carcinogenic to humans and is the causal factor of a rare aggressive skin malignancy the Merkel cell carcinoma (MCC) in around 80% of cases. Nevertheless, in addition to skin the virus was detected in various body tissues. In spite of that an ideal in vitro model system is still lacking. Three MCPyV isolates from healthy human skin are studied in this diploma thesis. The isolates harbour mutations, one has mutations is its LT coding gene, one has a rearranged non-coding control region (NCCR), and one is identical to the reference MPCyV isolate R17b. In this diploma thesis the replication capacities of the isolates were studied with the emphasis on the impact of a duplication within the NCCR and mutations on MCPyV genome replication in vitro after transfection. First, the results show that the NCCR rearrangement and the mutations in the viral protein coding genes impact the MCPyV replication in the 293 cell line. Preliminary data suggest the NCCR rearrangement negatively influences the MCPyV replication but certain mutations in protein coding genes could increase the replication. Moreover, none of the isolates replicated in the 293T cell line. The next part of the thesis focused on the replication of the MCPyV...
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Molecular mechanisms of fibroblastoid cell phenotype transitions:dedifferentiation of myofibroblasts and influencing of invasiveness and metastasis of sarcoma
Kosla, Jan
Fibroblasts are the principal cellular component of the connective tissue. They are a heterogeneous group of cells which contribute to the structure of connective tissue and wound healing by their ability to produce extracellular matrix (ECM). Fibroblasts and cells derived from them are involved in many pathological processes such as formation of malignant tumors and fibrosis. Tumor progression which finally leads to metastasis is a serious biomedical problem. There is a growing body of the recent evidence showing an important role of the tumor stroma and its interaction with cancer cells in cancer progression. Tumor stroma comprises mainly of myofibroblasts and their products, namely ECM, soluble factors, and enzymes. Myofibroblasts contribute more or less to all steps of cancer progression. Furthermore myofibroblasts play a key role in fibrosis, another serious human disease which is not efficiently treatable and which is associated with cancer progression. These facts made us to search for molecular means capable of eliminating the myofibroblastic phenotype. We succeeded to entirely dedifferentiate primary myofibroblasts by concomitant inhibition of TGFβ signaling and perturbation of MAPK signaling in a chick model that we have introduced. Malignant fibroblasts form sarcomas. ECM is the first...
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The role of new profibrotic molecules in the pathogenesis of systemic sclerosis.
Šumová, Barbora ; Šenolt, Ladislav (advisor) ; Funda, David (referee) ; Soukup, Tomáš (referee)
Systemic sclerosis (SSc) is immune-mediated fibrotic disease of unknown aetiology. Among the dominant pathogenic manifestations of SSc belong vascular changes, production of autoantibodies, activation of innate and adaptive immune responses and fibrotic processes. Transforming growth factor beta (TGF-β) has been identified as a central profibrotic factor stimulating fibroblasts to produce collagen. There are, however, a number of other mediators involved in the pathogenesis of SSc. Mutual activation and amplification of these molecules and their cascades may be a central mechanism of the SSc pathogenesis. Hedgehog (Hh) canonical signalling pathway plays an important role in the development and progression of fibrotic diseases. Expression of Hh target genes can be regulated through a canonical or non-canonical signalling cascade. The non-canonical activation of GLI transcription factors by TGF-β has not yet been investigated in SSc. The substantial part of this thesis is focused on the study of the mutual interaction of TGF-β and Hh signalling pathway. In vitro analysis confirmed TGF- β/SMAD3 dependent activation of GLI2 in dermal fibroblasts. Fibroblasts specific knockout of GLI2 prevented the development of experimental fibrosis in vivo. Combined targeting of canonical and non-canonical Hh...
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Molecular mechanisms of fibroblastoid cell phenotype transitions:dedifferentiation of myofibroblasts and influencing of invasiveness and metastasis of sarcoma
Kosla, Jan
Fibroblasts are the principal cellular component of the connective tissue. They are a heterogeneous group of cells which contribute to the structure of connective tissue and wound healing by their ability to produce extracellular matrix (ECM). Fibroblasts and cells derived from them are involved in many pathological processes such as formation of malignant tumors and fibrosis. Tumor progression which finally leads to metastasis is a serious biomedical problem. There is a growing body of the recent evidence showing an important role of the tumor stroma and its interaction with cancer cells in cancer progression. Tumor stroma comprises mainly of myofibroblasts and their products, namely ECM, soluble factors, and enzymes. Myofibroblasts contribute more or less to all steps of cancer progression. Furthermore myofibroblasts play a key role in fibrosis, another serious human disease which is not efficiently treatable and which is associated with cancer progression. These facts made us to search for molecular means capable of eliminating the myofibroblastic phenotype. We succeeded to entirely dedifferentiate primary myofibroblasts by concomitant inhibition of TGFβ signaling and perturbation of MAPK signaling in a chick model that we have introduced. Malignant fibroblasts form sarcomas. ECM is the first...
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Molecular mechanisms of fibroblastoid cell phenotype transitions:dedifferentiation of myofibroblasts and influencing of invasiveness and metastasis of sarcoma
Kosla, Jan ; Dvořák, Michal (advisor) ; Peková, Soňa (referee) ; Reiniš, Milan (referee)
Fibroblasts are the principal cellular component of the connective tissue. They are a heterogeneous group of cells which contribute to the structure of connective tissue and wound healing by their ability to produce extracellular matrix (ECM). Fibroblasts and cells derived from them are involved in many pathological processes such as formation of malignant tumors and fibrosis. Tumor progression which finally leads to metastasis is a serious biomedical problem. There is a growing body of the recent evidence showing an important role of the tumor stroma and its interaction with cancer cells in cancer progression. Tumor stroma comprises mainly of myofibroblasts and their products, namely ECM, soluble factors, and enzymes. Myofibroblasts contribute more or less to all steps of cancer progression. Furthermore myofibroblasts play a key role in fibrosis, another serious human disease which is not efficiently treatable and which is associated with cancer progression. These facts made us to search for molecular means capable of eliminating the myofibroblastic phenotype. We succeeded to entirely dedifferentiate primary myofibroblasts by concomitant inhibition of TGFβ signaling and perturbation of MAPK signaling in a chick model that we have introduced. Malignant fibroblasts form sarcomas. ECM is the first...
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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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