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Targeted differentiation of mesenchymal stem cells and their clinical application
Hámor, Peter ; Kubinová, Šárka (advisor) ; Trošan, Peter (referee)
The goal of this work is to point out possibilities of neuronal differentiation of mesenchymal stem cells and their application for clinical purposes, primary for repairing and regeneration of central nervous system tissues. Because this system works as a control center for functional features of the whole body, and treating this injuries and degenerations often bring many problems and obstacles, the possibility of using autologous cells for a transplantation or inducer of the natural regenerative properties of tissues is worth deeper research. This work progressively focuses on basic characteristics of stem cells and their differentiation potential, characterizing further mesenchymal stem cells together with possibilities of their isolation and cultivation. The main part of the text is formed by studies and methods used for targeted differentiation of mesenchymal stem cells and attempts of their transdifferentiation into neural cell line, together with present and possible future application of these cells in central nervous system therapies.
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Tracing the fate of cell populations from regressive tooth primordia during ontogenesis
Řadová, Marie ; Hovořáková, Mária (advisor) ; Černý, Jan (referee)
(v anglickém jazyce) Development of tooth primordia in mice is an important model for study of odontogenesis. Several dental rudiments develop during the mouse embryogenesis. These structures develop in functional teeth in their phylogenetically older relatives. Similarly, we can initiate growth of teeth from these germs in some mutant mice. In my diploma thesis we have focused on the importance of rudimentary structures with odontogenic potential in postnatal individuals. As a model of development, we have chosen a cell population originating from rudimentary primordia MS (mesial segment) that develops in diastema of the lower jaw during the embryonic day 12.5. Using the inducible Cre-lox technology we have marked the cells which are part of the signal domain of primordia at this time. As a marker of these cells we have used gene Shh. We have found out that these cells persist prenataly and also postnatally. Further we have isolated this cell area and we have tested it using a variety of methods. We have shown that in the cells of postnatal individual are expressed markers of stem cells (Sox2, Bmi1, Gli1) and also genes for major enamel matrix structural proteins: ameloblastin and amelogenin. The same stem cell markers are also expressed in vitro culture of the isolated cells. This cell population...
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3D bioprinting of stem cells and analysis of microscopic images
Kandra, Mário ; Svoboda, Ondřej (referee) ; Kolářová, Jana (advisor)
In this diploma thesis we are discussing about using 3D bioprinting in tissue engineering. We are discribing using biomaterials for construction scaffolder and aplication stem cells in 3D bioprinting. Last section of theoretical part deals with very often used techniques of 3D bioprinting and we are focused on extrusion technique. In the practical part we propose a method for print vasculars structures. We realized prototype of print head, her design and 3D printing of individual parts. To mechanical part we create a control system for printing control. At the end we visualize the organization of the cells using program modules.
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Differentiation of adult stem cells into insulin-producing beta cells
Koblas, Tomáš ; Saudek, František (advisor) ; Grim, Miloš (referee) ; Štechová, Kateřina (referee)
Ph.D. Thesis abstract: Diabetes mellitus is a chronic disease characterized by a metabolic disorder in which there is a low level or complete lack of the insulin. Diabetes mellitus type 1 (DM1) is caused by an autoimmune reaction leading to the destruction of the insulin producing beta cells in the pancreas. In consequence, low or non-existent insulin production leads to a complete dependence on exogenous insulin supplementation. DM1 causes serious long-term complications. Although strict control of blood sugar could prevent the onset and development of diabetic complications only 5% of diabetic patients are able to achieve such control. Hence it is evident that the current methods of treatment are neither sufficient to treat this disease, nor prevent late complications in most patients. The most promising therapeutic approach in the treatment of diabetes is the restoring of insulin production. One such method is the transplantation of insulin-producing tissue. However, a lack of available insulin- producing tissue limits such therapeutic approach. Therefore an alternative source of insulin producing cells have to be found to obtain a sufficient amount of safe and efficient insulin producing tissue. Pancreatic stem/progenitor cells could represent such an available alternative source. Despite the evidence...
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Stem cells in somatic tissues of mammals - past and present view
Hlavatá, Adéla ; Kořínek, Vladimír (advisor) ; Krylov, Vladimír (referee)
The homeostatic self-renewal of tissues in the adult mammal organism is maintained by stem- cell activity. The majority of tissue-specific stem cells are dispersed in a tissue in a low number. The small intestinal epithelium is a suitable model tissue for study of the stem cells because of its regular structure and rapid self-renewal. One of the first knowledge about the intestinal stem cell characteristics was obtained from the experiments with the mice embryonal aggregation chimeras and the transgenic chimeras. There were obtained a crucial role of the Wnt signalization pathway in the control of dynamics of the stem cells and contextual coherent finding of the unique molecular marker protein of the intestinal stem cells Lgr5. Subsequently, the genetically modified mice with "reporter" genes produced from the Lgr5 locus became the main tool in the intestinal epithelial stem cell research. The views on dividing organization and particular identity of the intestinal stem cells are still controversial. keywords: stem cells, crypt base cells, intestinal epithelium, chimera, the Wnt signaling pathway, Lgr5, reporter mouse strains
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Potential and significance of extended embryo culture
Uher, Petr ; Rokyta, Zdeněk (advisor) ; Slípka, Jaroslav (referee) ; Pilka, Ladislav (referee)
The aim of this dissertation thesis is to emphasize the sense of extended cultivation of embryos to the stadium of blastocyst and its influence on success of assisted reproduction and facilitation of pre implantation diagnosis, analysis of cultivation media and derivation of human embryonic stem cells. Author summarizes current literary findings in assisted reproduction and examines the currently used methods. Author also submits his own published experimental works, in which he compares his own results of infertility treatment with usage of extended cultivation to blastocyst with results of other techniques. Furthermore author submits his own published experimental works which are using extended cultivation for pre implantation diagnosis and its improvement. Another experimental works includes possibility of stem cells derivation. Usage of extended cultivation to blastocyst convincingly leads, according to author's own experiments and simultaneously to available literary findings, to higher success of infertility treatment. This is especially significant by middle-aged mothers. Sufficient term of cultivation enables not just selection, but also biopsy and its generic treatment. Long-term cultivation also enables analysis of cultivation media - but these didn't met the expectations for increase of...
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Cell therapy in animal models - preclinical studies
Juhásová, Jana ; Motlík, Jan (advisor) ; Grim, Miloš (referee) ; Jendelová, Pavla (referee)
The progress of cell therapy can be greatly facilitated by using suitable experimental models. It is essential to verify the clinical usefulness of new healing procedures obtained in studies on laboratory animals by using a large animal model. One of suitable models well acceptable in medical community is undoubtedly the miniature pig, which resembles humans in terms of physiology and body proportions. This PhD thesis presents the summary of our experimental studies relating to possible exploitation of mesenchymal and neural stem cells in the healing of locomotive apparatus and neural tissue disorders in humans or animals. The first part of the thesis briefly describes the current issue of cell therapy and animal models, mesenchymal cells and/or their combination with new types of scaffolds, neurogenesis, neural stem cells and their potential application in therapy of spinal cord injury. The second part is focused on the goals and methodology, the individual publications being listed in the third part. Our experiments with iatrogenic physeal defect in rabbits, which served as a model of the occurrence of valgous deformation in the clinical practice, showed the positive preventive and therapeutical effects of a new type of scaffolds seeded with allogeneic mesenchymal stem cells in animals without...
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Experimental models of a transfer of stem cells for therapeutic purposes
Faltýsková, Helena ; Krulová, Magdaléna (advisor) ; Indrová, Marie (referee)
Experimental models of a transfer of stem cells for therapeutic purposes Abstract Stem cell therapy currently represents a standard procedure of treating a wide variety of hereditary diseases and serious injuries. Development of the most suitable way of transfer of stem cells into the patient body remains very important question concerning this type of therapy. In our experiments we used nanofiber scaffolds for stem cell cultivation and their subsequent transfer. These nanofibers were prepared by the original needleless electrospun NanospiderTM technology. Allogeneic cornea or skin graft were transplanted from B6 mice to BALB/c mice. The grafts were covered by a nanofibrous scaffold with cultivated stem cells. Stem cells were stained by an imunofluorescent dye to enable us to monitore their migration from nanofibers into tissues and consequent distribution in the body and characterize changes of this distribution in the time. The methods of ELISA and PCR were used to confirm that mesenchymal stem cells support the production of antiinflammatory cytokines IL-4 and IL-10 and contribute to inhibition of production of proinflammatory cytokines IL-1, IFNγ and inducible nitric oxide synthase. We confirmed an important beneficial role of nanofiber scaffolds in transplantation of mesenchymal stem cells. Nanofiber...
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Morphological variability of the incisor in mutant mice
Lochovská, Kateřina ; Churavá, Svatava (referee) ; Hovořáková, Mária (advisor)
Myš je nejfrekventovaněji používaným experimentálním modelovým organismem pro studium vývoje zubů. Myší funkční dentice obsahuje jeden řezák oddělený od tří molárů dlouhou bezzubou diastemou v každém čelistním kvadrantu. Cílem této práce bylo shrnout poznatky o myší dentici a jejím vývoji se zaměřením na myší řezáky a jejich patologie. Myší řezák je díky svým vlastnostem jedinečným zubem. Charakteristickou vlastností hlodavců jsou právě kontinuálně rostoucí řezáky. Tyto řezáky jsou pokryté sklovinou pouze na labiální straně. Lingvální povrch je tvořen pouze dentinem. Toto je spojeno s asymetrickou abrazí. Nicméně je také častým cílem mutací, ať už v podobě delece nebo jiné modifikace genů. Tyto mutace dentálních signálních drah jsou studovány na mutantních myších jako jsou například Tabby myši, Sprouty nebo Small eye (Sey) mutantní myši a mnoho dalších. Některé mutace jsou homologní k lidským onemocněním. Například X-vázaný tabby (Ta) syndrom u myší je považován za homolog hypohidrotické ektodermální dysplásie (HED) u lidí. Tato mutace napadá velikost řezáku, jeho tvar a pozici stejně jako cytodiferenciaci. Dále se může objevit hypodoncie, anodoncie nebo nějaké morfologické změny napadající ostatní existující zuby. Abnormality v počtu, velikosti a tvaru zubu byly dobře zdokumentovány v lidské...
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