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Simulation of adhesion and transmigration imune-cells through capillary wall
Morgaenko, Katsiarina ; Čmiel, Vratislav (oponent) ; Chmelíková, Larisa (vedoucí práce)
Theoretical part of this study includes biophysical description of endothelial layer and cell transmigration through it. There is descripted characteristics of all important components used for creation of capillary model in vitro (endothelial cells, stem cells, white blood cells, Calcein AM, PKH26). Practical part of the study devotes to fabrication of two-dimensional and three-dimensional in-vitro models of endothelial layers, processing of visualization by microscope and testing interaction between leukocytes, stem cells and endothelial layer. In conclusion is proposed the best form and size of microfluidic models simulated capillary.
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The cell migration monitoring in a microfluidic system by the "Scratch Wound Healing Assay" method
Morgaenko, Katsiarina ; Skopalík, Josef (oponent) ; Chmelíková, Larisa (vedoucí práce)
This thesis describes the cell culture methods of mouse embryonic fibroblast (3T3), human umbilical vein endothelial (HUVEC), and Chinese hamster ovary (CHO) cell lines in microfluidic systems simulating capillary. Literature research for microfluidic realizations of Scratch Wound Healing Assay with fibroblasts and endothelial cell lines was performed in this thesis. The principles of confocal and fluorescence microscopy, and relevant image processing techniques used for quantifying cell migration were studied. Experimental setup for microfluidic Scratch Wound Healing Assay with using of trypsin – EDTA for creation of cell delusion, and confocal microscope Leica TCS SP8 X for subsequent image acquisition was proposed and tested. An appropriate algorithm for cell migration analysis was written in Matlab computing environment. Experiment’s results are discussed in the conclusion of this thesis.
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The cell migration monitoring in a microfluidic system by the "Scratch Wound Healing Assay" method
Morgaenko, Katsiarina ; Skopalík, Josef (oponent) ; Chmelíková, Larisa (vedoucí práce)
This thesis describes the cell culture methods of mouse embryonic fibroblast (3T3), human umbilical vein endothelial (HUVEC), and Chinese hamster ovary (CHO) cell lines in microfluidic systems simulating capillary. Literature research for microfluidic realizations of Scratch Wound Healing Assay with fibroblasts and endothelial cell lines was performed in this thesis. The principles of confocal and fluorescence microscopy, and relevant image processing techniques used for quantifying cell migration were studied. Experimental setup for microfluidic Scratch Wound Healing Assay with using of trypsin – EDTA for creation of cell delusion, and confocal microscope Leica TCS SP8 X for subsequent image acquisition was proposed and tested. An appropriate algorithm for cell migration analysis was written in Matlab computing environment. Experiment’s results are discussed in the conclusion of this thesis.
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Simulation of adhesion and transmigration imune-cells through capillary wall
Morgaenko, Katsiarina ; Čmiel, Vratislav (oponent) ; Chmelíková, Larisa (vedoucí práce)
Theoretical part of this study includes biophysical description of endothelial layer and cell transmigration through it. There is descripted characteristics of all important components used for creation of capillary model in vitro (endothelial cells, stem cells, white blood cells, Calcein AM, PKH26). Practical part of the study devotes to fabrication of two-dimensional and three-dimensional in-vitro models of endothelial layers, processing of visualization by microscope and testing interaction between leukocytes, stem cells and endothelial layer. In conclusion is proposed the best form and size of microfluidic models simulated capillary.
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