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Study of mesenchymal stem cell migration in the extracellular matrix based on principles of chemotaxis
Scholasterová, Viktorie ; Fohlerová, Zdenka (referee) ; Chmelíková, Larisa (advisor)
This thesis engages in a study of mesenchymal stem cell migration in extracellular matrix based on principles of chemotaxis. First, attention is focused on a theoretical part associated with a clarification of basic terms such as extracellular matrix, migration, confocal microscopy, mesenchymal stem cells or chemotaxis. There is also included a list and a description of some basic methods for monitoring cell migration and a more detailed description of a method called transwell assay, which has been chosen for an experiment in a practical part of this thesis. This part includes protocols of individual steps for the preparation of the experiment, the procedure of data processing obtained by scanning cells with a confocal microscope and a description of the resulting confluence values.
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Study of mesenchymal stem cell migration based on principles of chemotaxis
Pošustová, Veronika ; Skopalík, Josef (referee) ; Chmelíková, Larisa (advisor)
The purpose of this Master thesis is to verify migration of mesenchymal stem cells on the principle known as chemotaxis. First part of this study is focused on cell migration in order to explain the whole migration process. Next part describes various chemotaxis methods and selected studies dealing with clinical applications of mesenchymal stem cells in different medical and biomedical fields. The following step describes confocal microscopy, which is used for acquiring images of the cells. The experimental part is focused on cultivation of mesenchymal stem cells in a laboratory, which is necessary for cell vitality. Furthermore, there are designed two main experiments. Firstly there is a 2D experiment with adherent cells for chemotaxis using -Slide Chemotaxis. Secondly Transwell migration test is designed and executed. Finally, the acquired images from confocal microscope are used for image processing, which was done in Matlab R2020a programming environment. The result of this processing is evaluation of cell confluence and migration. In the end, experimental part of this study was optimized according to recommended studies. The results are summarized in the conclusion with proposal for improvements of those methods.
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Chemical communication of gametes
Otčenášková, Tereza ; Komrsková, Kateřina (advisor) ; Petr, Jaroslav (referee)
Fertilization is a multiple step process leading to fusion of female and male gametes resulting in a formation of a zygote. Besides direct gamete interaction via binding receptors localized on both oocyte and sperm surface, fertilization also involves communication based on chemical molecules triggering various signalling pathways. This work is aimed to characterize chemical communication of gametes of a model organism Mus musculus. For this purpose, modern proteomic and visualisation methods like nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS), selected reaction monitoring (SRM) and immunofluorescent microscopy were used. Lipocalins were identified as candidate proteins involved in communication including those from major urinary proteins (MUPs), LCN lipocalins and fatty acid binding proteins (FABPs). For the first time, we report their presence in the sperm acrosome. Based on lipocalins capacity to bind and transport other molecules, we propose that these proteins have a protective and/or signalling role for gametes. Furthermore, chemical communication between sperm and oocyte is based on chemotaxis which enables their interaction before their fusion. In this work, we detected that spermatozoa show chemotactic responses in the presence of L-glutamate. This amino acid naturally...
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Study of mesenchymal stem cell migration in the extracellular matrix based on principles of chemotaxis
Scholasterová, Viktorie ; Fohlerová, Zdenka (referee) ; Chmelíková, Larisa (advisor)
This thesis engages in a study of mesenchymal stem cell migration in extracellular matrix based on principles of chemotaxis. First, attention is focused on a theoretical part associated with a clarification of basic terms such as extracellular matrix, migration, confocal microscopy, mesenchymal stem cells or chemotaxis. There is also included a list and a description of some basic methods for monitoring cell migration and a more detailed description of a method called transwell assay, which has been chosen for an experiment in a practical part of this thesis. This part includes protocols of individual steps for the preparation of the experiment, the procedure of data processing obtained by scanning cells with a confocal microscope and a description of the resulting confluence values.
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Study of mesenchymal stem cell migration based on principles of chemotaxis
Pošustová, Veronika ; Skopalík, Josef (referee) ; Chmelíková, Larisa (advisor)
The purpose of this Master thesis is to verify migration of mesenchymal stem cells on the principle known as chemotaxis. First part of this study is focused on cell migration in order to explain the whole migration process. Next part describes various chemotaxis methods and selected studies dealing with clinical applications of mesenchymal stem cells in different medical and biomedical fields. The following step describes confocal microscopy, which is used for acquiring images of the cells. The experimental part is focused on cultivation of mesenchymal stem cells in a laboratory, which is necessary for cell vitality. Furthermore, there are designed two main experiments. Firstly there is a 2D experiment with adherent cells for chemotaxis using -Slide Chemotaxis. Secondly Transwell migration test is designed and executed. Finally, the acquired images from confocal microscope are used for image processing, which was done in Matlab R2020a programming environment. The result of this processing is evaluation of cell confluence and migration. In the end, experimental part of this study was optimized according to recommended studies. The results are summarized in the conclusion with proposal for improvements of those methods.
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Chemical communication of gametes
Otčenášková, Tereza ; Komrsková, Kateřina (advisor) ; Petr, Jaroslav (referee)
Fertilization is a multiple step process leading to fusion of female and male gametes resulting in a formation of a zygote. Besides direct gamete interaction via binding receptors localized on both oocyte and sperm surface, fertilization also involves communication based on chemical molecules triggering various signalling pathways. This work is aimed to characterize chemical communication of gametes of a model organism Mus musculus. For this purpose, modern proteomic and visualisation methods like nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS), selected reaction monitoring (SRM) and immunofluorescent microscopy were used. Lipocalins were identified as candidate proteins involved in communication including those from major urinary proteins (MUPs), LCN lipocalins and fatty acid binding proteins (FABPs). For the first time, we report their presence in the sperm acrosome. Based on lipocalins capacity to bind and transport other molecules, we propose that these proteins have a protective and/or signalling role for gametes. Furthermore, chemical communication between sperm and oocyte is based on chemotaxis which enables their interaction before their fusion. In this work, we detected that spermatozoa show chemotactic responses in the presence of L-glutamate. This amino acid naturally...
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Orientation of helminths during host seeking
Vojtová, Terezie ; Mikeš, Libor (advisor) ; Horák, Petr (referee)
The parasitic helminths are often forced in the environment to search for their host quickly in order to continue the ontogenetic development, or else to finish it. To search for and indentify the host, they use various types of receptors which allow them to provoke a taxes corresponding to the oncoming stimulus and to react regarding the current situation. In respect to the concept relevant to the helminths, this work concerns trematodes, whose freely movable stadia are miracidium and cercacia, and then monogeneans which are viparous or oviparous with oncomiracidium larva, and nematodes during which the third larva stage moves. The representatives of the groups of Myxozoans and Ciliophorea are also mentionned, since they use similar mechanisms to look for hosts as helminths. This thesis includes the summary of the existing knowledge related to the orientation of helminths in the environment. On the basis of the data that is already known, it is possible to compare which mechanisms are used for host-seeking by particular species, and according to which signals from the surroundings they most often orientate themselves. Powered by TCPDF (www.tcpdf.org)
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Orientation of helminths during host seeking
Vojtová, Terezie ; Mikeš, Libor (advisor) ; Horák, Petr (referee)
Helminths are often forced in the environment to search for their host quickly in order to continue the ontogenetic development, or else to finish it. To search for and indentify the host, they use various types of receptors which allow them to provoke a taxes corresponding to the oncoming stimulus and to react to the presence of the host. This thesis includes the summary of the existing knowledge related to the orientation of helminths, the comparison of the mechanisms used by the searching of a host and signals in the environment. In respect to the concept relevant to the helminths, this work concerns trematodes, whose freely movable stadia are miracidium and cercaria, and then monogeneans which are viparous or oviparous with oncomiracidium larva, and nematodes with the third infective larva stage. The representatives of fish parasites of Myxozoans and Ciliophorea are also mentionned, since they use similar mechanisms to look for hosts as helminths. Powered by TCPDF (www.tcpdf.org)
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